Skip to main content
PMC home page
Wiley Open Access Collection logoLink to Wiley Open Access Collection
. 2020 Nov 9;148(6):1489–1498. doi: 10.1002/ijc.33370

Short tandem repeat profiling for the authentication of cancer stem‐like cells

Paola Visconti 1, Federica Parodi 1,✉, Barbara Parodi 1, Lucia Casarino 2, Paolo Romano 3, Mariachiara Buccarelli 4, Roberto Pallini 5, Quintino Giorgio D'Alessandris 5, Andrea Montori 6, Emanuela Pilozzi 6, Lucia Ricci‐Vitiani 4
PMCID: PMC7894552  PMID: 33128777

Abstract

Colorectal and glioblastoma cancer stem‐like cells (CSCs) are essential for translational research. Cell line authentication by short tandem repeat (STR) profiling ensures reproducibility of results in oncology research. This technique enables to identify mislabeling or cross‐contamination of cell lines. In our study, we provide a reference dataset for a panel of colorectal and glioblastoma CSCs that allows authentication. Each cell line was entered into the cell Line Integrated Molecular Authentication database 2.1 to be compared to the STR profiles of 4485 tumor cell lines. This article also provides clinical data of patients from whom CSCs arose and data on the parent tumor stage and mutations. STR profiles and information of our CSCs are also available in the Cellosaurus database (ExPASy) as identified by unique research resource identifier codes.

Keywords: cell line authentication, colorectal tumor, glioblastoma, human stem‐like cell lines, short tandem repeat profiling

What's new?

Human cell lines obtained from cancer stem‐like cells represent an invaluable model for studying tumor properties. Cell line authentication by short tandem repeat (STR) profiling is an important tool to identify the potential mislabeling or cross‐contamination of cell lines. Here, the authors characterized 18 colorectal cancer stem‐like cell lines from 17 patients and 103 glioblastoma cancer stem‐like cell lines from 95 patients by STR profiling to create a reference dataset that allows the authentication of these cell lines and their identification through a unique research resource identifier. The results will help further ensure the reliability and reproducibility of research experiments.

Abbreviations

AMELX

amelogenin in Xp22.1‐22.3

AMELY

amelogenin in Yp11.2

CLASTR

cellosaurus STR database

CLIMA

Cell Line Integrated Molecular Authentication database

CSC

cancer stem‐like cell

CTSC

colorectal tumor stem‐like cell

GSC

glioblastoma stem‐like cell

ICLAC

International Cell Line Authentication Committee

ICLC

Cell Bank Interlab Cell Line Collection

MGMT

O6‐methylguanine DNA methyltransferase

MSI

microsatellite instability

PBMC

peripheral blood mononuclear cell

PCR

polymerase chain reaction

STR

short tandem repeat

1. INTRODUCTION

Cancer research requires models that use patient‐derived cultured cells. These models allow to study tumor heterogeneity, in particular in the early stages of tumorigenesis. Isolation and in vitro cultivation of cancer stem‐like cells (CSCs), a small fraction of self‐renewing cells with stem‐like properties, provide a model to study the properties of these tumor initiating cells. 1 , 2 , 3 , 4 , 5 , 6 Stem‐like cells are grown as free‐floating oncospheres in serum‐free medium supplemented with growth factors. 1 , 2 , 3 , 4 , 5 , 6 When grafted as orthotopic models, CSCs closely reproduce the parent tumor, both histologically and genetically. Therefore, the identification of each cell line is essential in oncology research. 7 , 8 , 9 Short tandem repeat (STR), a standard authentication technique that allows identification of the individual from whom each cell line originates, 10 amplifies a set of polymorphic STR markers and then separates the polymerase chain reaction (PCR) products by capillary electrophoresis size fractionation. We characterized 18 colorectal CSC lines (CTSCs) 11 from 17 patients and 103 glioblastoma CSCs (GSCs) 3 from 95 patients by STR assay to create a reference dataset that allows the determination of the authenticity of these cell lines and ensures the reliability and reproducibility of research experiments. In one patient with a colorectal tumor and in seven patients with glioblastoma, we established CSC lines from different portions of the same tumor. Moreover, in two glioblastoma patients, of whom we obtained GSC lines from both primary surgery and surgery for recurrence, the STR profile confirmed the authenticity of CSC lines derived from the same patient. Peripheral blood mononuclear cells (PBMCs) and primary tumor cells of some patients were also analyzed. Due to the availability of the surgical samples, we could preserve tumor tissue for genotyping only in a minority of patients.

STR profiling was carried out using standardized procedures for unambiguous authentication and identification of human cell lines according to the American National Standards Institute/American Type Culture Collection Standard ASN‐0002‐2011. 12 Each cell line profile presented in this artcle was entered in a specific data set (ICLC 3) of the Cell Line Integrated Molecular Authentication database (CLIMA), 13 including STR profiles obtained in different cell banks by using different platforms. All STR profiles were also compared using the cellosaurus STR database (CLASTR) search tool of the Cellosaurus database (ExPASy) (https://web.expasy.org/cellosaurus/).

The cell line profiles were challenged against public databases to exclude cross‐contamination with commercially available cell lines. Comparison of the cell line profiles against each other ruled out duplicates due to cross‐contamination. Duplicates were found only in those cell lines that derived from different regions of the same tumor.

2. MATERIALS AND METHODS

Further method descriptions are included in Supplementary Material and Methods.

2.1. CSC cultures

CTSCs and GSCs were isolated from tumor samples through mechanical dissociation and cultured in a serum‐free medium supplemented with growth factors, as previously described. 1 , 2 , 3 , 4 , 5 , 6 Under stem cell culture conditions, proliferating CSC lines actively required 3 to 4 weeks to be established (Supplementary Material and Methods).

2.2. Mycoplasma statement

All experiments were performed with mycoplasma‐free cells. Mycoplasma contamination in cell cultures was evaluated using MycoAlert Mycoplasma Detection Kit (Lonza Walkersville Inc, Walkersville, MD).

2.3. Molecular analyses in CTSCs and colorectal tumors

Single‐point mutations and small insertions/deletions in CTSCs were assessed by targeted DNA resequencing, focusing on 17 genes known to be frequently mutated in colon cancer, as previously described. 11

Microsatellite instability (MSI) detection was performed using the Promega panel of mononucleotide MSI markers (MSI Analysis System, Version 1.2, Promega Corporation, Fitchburg, WI). The expression of four mismatch repair proteins, MLH1, MSH2, MSH6 and PMS2, was investigated in tumor tissues corresponding to CTSCs that showed MSI‐High.

The expression of the stem cell marker CD133 and the epithelial antigen Ber‐EP4 in CTSCs was evaluated by flow cytometry, as previously described. 5

2.4. Molecular analyses in GSCs and glioblastoma tissues

Tumor proliferation index was analyzed by immunohistochemistry on paraffin sections of glioblastoma samples using the avidin‐biotin‐peroxidase complex methods (ABC‐Elite kit, Vector, Burlingame, CA). 14 The anti‐Ki‐67 monoclonal antibody (MIB‐1, Dako) was used. The expression of CD133 and Sox2 in GSCs was evaluated by flow cytometry. O6‐methylguanine DNA methyltransferase (MGMT) promoter methylation patterns were studied by methylation‐specific PCR on genomic DNA. 15 DNA from normal lymphocytes treated with SssI methyltransferase (New England Biolabs, Ipswich, MA) was used as positive control. PCR products were separated onto 3% agarose gel, stained with ethidium bromide and visualized under UV illumination.

2.5. STR profiling

All human cell lines described have been authenticated by STR profiling within the last 3 years. Genomic DNA was isolated from cell pellets using the DNeasy Blood & Tissue Kit (Qiagen, Milan, Italy) and treated with RNase, according to the manufacturer's instructions. Yield was measured with NanoDrop 1000 spectrophotometer (Thermo Fisher Scientific, Wilmington, DE). One nanogram of DNA of each sample was used for the STR analysis. Samples were amplified and electrophoretically separated on an ABI Prism 3100 Genetic Analyzer (Applied Biosystems).

STR profiles were analyzed by the GeneMapper ID software (Applied Biosystems), Version 3.2. The results showed highly reproducible cell line‐specific numeric patterns. The assay was performed by the Cell Bank Interlab Cell Line Collection (ICLC) of the Biological Resource Centre, IRCCS Ospedale Policlinico San Martino of Genoa, in collaboration with the Department of Legal and Forensic Medicine of Genoa University. Comparison of STR profiles using the CLIMA database was performed using the identification feature in CLIMA 2.1 version of the database, as previously described. 13 Each new cell line profile was compared to profiles of all cell lines (4485 cell lines names and 5587 distinct authentication assays) contained in the database, that are divided into datasets (Table S1). All STR profiles of CTSCs and GSCs were compared using CLASTR, the Cellosaurus STR similarity search tool that contains more than 6400 distinct cell lines with an associated STR profile. 16

3. RESULTS

3.1. Patient clinical data and molecular cell line characterization

We characterized 18 CTSCs from 17 patients (7 males and 10 females, mean age 68.4 years). Tumor site and disease stage (grade, pTNM and Dukes) of CTSCs are shown in Table 1. Mutations were harbored by CTSCs in the following genes: ACVR1B, AMER1, APC, BRAF, CTNNB1, FBXW7, KIAA1804, KRAS, MAP2K4, NRAS, PIK3CA, PTEN, SMAD2, SMAD4, SOX9, TCF7L2 and TP53 (Table 1; Table S2).

TABLE 1.

STR profiles, disease stage and principal mutations of CTSCs

Patient ID Sex Age Tumor site Disease grade Disease pTNM Disease dukes Cell line name Cellosaurus accession number ACVR1B AMER1 APC BRAF CTNNB1 FBXW7 KIAA1804 KRAS MAP2K4 NRAS PIK3CA PTEN SMAD2 SMAD4 SOX9 TCF7L2 TP53 STR ID AM D5S818 D13S317 D7S820 D16S539 VWA TH01 TPOX CSF1PO MSI Status Ref.
1 M 68 Left G3 IIIB C CTSC#1.1 CVCL_A9WH ■ ■ ■ ■ ■ 254 X,Y 11,13 8,9 10 9,11 16,17 8,9 8 12 MSS 5, 11
1 M 68 Left G2 IIIB C CTSC#1.2 CVCL_A9WI ■ ■ ■ ■ ■ ■ 250 X,Y 11,13 8,9 10 9,11 16,17 8,9 8 12 MSS 5, 11
2 F 66 Right G2 IIA B CTSC#18 CVCL_A9WJ ■ ■ ■ ■ ■ ■ ■ ■ ■ 248 X 11,12 11,12 10,13 11 17 6,9.3 8,9 9,14 MSI‐H 5, 11
3 F 63 Right G2 IVA D CTSC#CRO CVCL_A9WK 251 X 11,12 11,14 10,11 9,11 17,19 7,9.3 8 10 MSS 11
4 M 64 Right G2 IIA B CTSC#85 CVCL_A9WL ■ ■ ■ ■ 249 X,Y 9,12 8,12 10 11,13 14 6,7 8 10,12 MSS 11
5 M 80 Left G3 IIIC C CTSC#383 CVCL_A9WM ■ ■ ■ ■ ■ 413 X,Y 12 9,11 11,12 12,15 17,18 9,9.3 9,11 10,11 MSS 11
6 M 76 Right G2 IIIC C CTSC#389 CVCL_A9WN ■ ■ ■ ■ ■ ■ 500 X,Y 12,13,15 14,15 8,11 9,14 15,18,19 9,9.3 10,12 10,12 MSI‐H 11
7 M 57 Right G3 IIIC C CTSC#393 CVCL_A9WT ■ ■ ■ ■ 416 X,Y 12 9,12 12 9 18 7,9 8,11 13 MSS 6 , 11
8 F 46 Left G2 IIIB C CTSC#398 CVCL_A9WQ ■ ■ ■ ■ ■ 415 X 12 8,10 8,10 9,14 14,15 6 10,12 11 MSS 6 , 11
9 M 82 Right G3 IIIC C CTSC#416 CVCL_A9WR ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ 501 X,Y 11,14 7,12 12,13 11,14 15,18 6,9.3 7 11,12,13 MSI‐H 6 , 11
10 F 70 Right G2 IIA B CTSC#417 CVCL_A9WS ■ ■ ■ 417 X 11 11 11 12 16,17 6,8 9 12 MSI‐L 11
11 F 68 Right G3 IIIB C CTSC#430 CVCL_A9WT ■ ■ ■ ■ ■ 498 X 11,15 8,11 8,14 11 16,19,20 9,9.3 9,11 11 MSI‐H 6 , 11
12 M 49 Left G3 IIIB C CTSC#432 CVCL_A9WU ■ ■ ■ ■ 499 X,Y 10,13 10,12 10,12 13 17,18 8,9.3 8 10,11 MSS 6 , 11
13 F 87 Right G3 IVB — CTSC#446 CVCL_A9WW ■ ■ ■ ■ ■ ■ 502 X 13 9,12 10,11 11,13 17 9.3 8,11 11 MSS
14 F 73 Right G3 IIA B CTSC#510 CVCL_A9WY ■ ■ ■ ■ ■ ■ ■ ■ ■ 606 X 12,13 9,12 10,11 9,12 18,2 8,9 8,9 10,11 MSS
15 F 85 Right G3 IIIC C CTSC#438 CVCL_A9WV ■ ■ ■ ■ ■ ■ ■ 579 X 11,14 9,12 9,10 9,10 16,17 6,9 8 11 MSI‐H 11
16 F 63 Right G3 IIIC C CTSC#482 CVCL_A9WX ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ ■ 708 X 9,10 11,12 9,10 10 15,16,17,18 6,9 8,9,10 12,13 MSI‐H
17 F 66 Right G3 IIIB C CTSC#553 CVCL_A9WZ — — — — — — — — — — — — — — — — — 604 X 11,12 11,12 11,12 9,12 14,17 9,10 8 12 MSS
Open in a new tab

Notes: STR profiles generated in our study from CTSCs. Tumor site and disease stage (grade, pTNM and dukes) of colorectal tumors that originated CTSCs and the principal mutations harbored by CTSCs are also shown. Mutation in the following genes were analyzed: ACVR1B, AMER1, APC, BRAF, CTNNB1, FBXW7, KIAA1804, KRAS, MAP2K4, NRAS, PIK3CA, PTEN, SMAD2, SMAD4, SOX9, TCF7L2, TP53 (details are shown in Table S2). Shown is also MSI status of CTSCs. MSI‐H, MSI‐high; MSI‐L, MSI‐low; MSS, microsatellite stable (details are shown in Table S3). Previous studies involving CTSCs are indicated by the number of reference in the last column of the table.

Abbreviations: F, female; M, male; STR, short tandem repeat; CTSC, colorectal tumor stem‐like cell.

Analysis of MSI status using mononucleotide MSI markers showed that 6 CTSC lines had MSI‐High (CTSC#18, CTSC#389, CTSC#416, CTSC#430, CTSC#438, CTSC#482), one line had MSI‐Low (CTSC#417) and the other lines were microsatellite stable (Table S3). In accordance, expression analysis of the four mismatch repair proteins, MLH1, MSH2, MSH6 and PMS2, assessed in tumor tissues corresponding to the microsatellite instable CTSCs, also showed MSI‐High (Table S3).

CTSCs were analyzed for the expression of CD133 and epithelial antigen Ber‐EP4 (Table S4). CD133 is one of the key stem cell markers for colorectal cancer 5 and its expression is associated with cell differentiation and tumor size. 17 In our collection of CTSCs, CD133 and Ber‐EP4 expression was 63.4% ± 6.4% (mean ± SEM; range 23.2%‐99.3%) and 94.8% ± 1.3% (mean ± SEM; range 82.8%‐100%), respectively.

We characterized 103 GSCs from 95 glioblastoma patients (66 males and 29 females, mean age 62.3 years). Tumor location, disease stage (primary/recurrent tumor), MGMT gene methylation status and proliferation index (Ki67) of parent tumors are shown in Table 2. MGMT gene promoter status was methylated in 46 tumors (49%), unmethylated in 44 tumors (46%) and not available in 5 tumors (5%).

TABLE 2.

STR profiles, disease stage, MGMT methylation and proliferation index Ki67 (%) of GSCs

Patient ID Sex Age Tumor location Type of tumor Cell line name Cellosaurus accession number CVCL MGMT Ki67 (%) STR ID AM D5S818 D13S317 D7S820 D16S539 VWA TH01 TPOX CSF1PO Ref.
19 m 40 Temporal pt GSC#1 CVCL_A9T7 M 20 418 X 12,13 8 8,10 11,14 16,18 6,9.3 8 10 1 , 2 , 3 , 4
20 m 57 Parietal pt GSC#10 CVCL_A9T8 M 30 631 X,Y 11 11,12 10,11 11,12 16,17 9.3 8 11,12 1 , 2 , 3 , 4
21 m 77 Parietal pt GSC#23C CVCL_A9T9 UM 50 419 X,Y 11 12 10,11 9,11 17 7 8,9 11,12 1 , 2 , 3 , 4
21 m 77 Parietal pt GSC#23P CVCL_A9TA UM 50 492 X,Y 11 12 10,11 9,11 17 7 8,9 11 1 , 2 , 3 , 4
22 m 72 Frontal pt GSC#28 CVCL_A9TB M 5 427 X,Y 10,12 10,11 9,10 10,14 14,16 9,9.3 8,11 10,11 1 , 2 , 3 , 4
23 m 44 Frontal pt GSC#30P CVCL_A9TC M 10 428 X,Y 11,12 12 10,12 12 14,16 6,9 8,11 11,12 1 , 2 , 3 , 4
23 m 44 Frontal pt GSC#30pt CVCL_A9TD M 10 493 X 11,12 12 10 12 14,16 6,9 8,11 11,12 1 , 2 , 3 , 4
24 m 59 Occipital pt GSC#61 CVCL_A9TE UM 35 420 X,Y 11 8,12 9,10 10,13 17 9.3 11 11 1 , 2 , 3 , 4
25 m 64 Frontal rt GSC#62 CVCL_A9TF M 10 421 X,Y 9 11,12 7,11 12 15,17 6,8 8 10,13 1 , 2 , 3 , 4
26 m 48 Parietal pt GSC#67 CVCL_A9TG UM 20 422 X,Y 10,13 11,13 8 9,11 18 6,7 11 10,12 1 , 2 , 3 , 4
27 m 58 Parietal pt GSC#68 CVCL_A9TH UM 10 423 X,Y 11,12 10,11 10,11 11,13 15,17 6,9.3 8 10 1 , 2 , 3 , 4
28 f 67 Parietal pt GSC#70 CVCL_A9TI UM 20 424 X 11,12 12,13 8,10 11,12 14,17 9 8,11 11 1 , 2 , 3 , 4
29 f 70 Frontal pt GSC#74 CVCL_A9TJ UM 15 425 X 11,12 8,12 9 11,12 15 9,9.3 8 10 1 , 2 , 3 , 4
30 f 48 Frontal pt GSC#76 CVCL_A9TK UM 15 426 X 10 12 8,12 12 17 7 11 12 1 , 2 , 3 , 4
31 m 52 Temporal pt GSC#83 CVCL_A9TL UM 40 445 X 11 12 12 10,11 17,18 6 8,11 10,11 1 , 2 , 3 , 4
31 m 52 Temporal pt GSC#83.2 CVCL_A9TM UM 40 446 X 11 12 12,13 10,11 17,18 6 8,11 10,11 1 , 2 , 3 , 4
32 f 49 Parietal pt GSC#112 CVCL_A9TN M 18 619 X 12 12 11 11 17 7 11 10 1 , 2 , 3
33 m 53 Parietal rt GSC#120 CVCL_A9TP UM 30 432 X,Y 11 11 10,11 9,11 14,21 9.3 8 10,13 1 , 2 , 3
34 m 57 Temporal pt GSC#142 CVCL_A9TQ M 40 429 X 10.13 9 8,10 12 16,17 6,9.3 8,11 11 1 , 2 , 3
35 f 69 Frontal pt GSC#147 CVCL_A9TR UM 25 440 X 12,13 11 8,12 9,12 15,17 6,8 8 11,12 1 , 2 , 3
36 m 55 Parietal rt GSC#148 CVCL_A9TS UM 70 433 X 9 10 8,10 12,13 18 6,9.3 8,9 9 1 , 2 , 3
37 m 69 Occipital pt GSC#151 CVCL_A9TT M 30 434 X,Y 13 12 9,11 11,12 16,18 7,9 11 12,13 1 , 2 , 3
38 m 56 Parietal pt GSC#163 CVCL_A9TU UM 15 442 X,Y 11,12 12 9,11 11 18 7,9 8 10,12 1 , 2 , 3
39 m 61 Temporal pt GSC#169 CVCL_A9TV UM 40 436 X,Y 11,12 9 10,11 11,13 14 10 11 10,11 1 , 2 , 3
40 m 58 Temporal — GSC#170 CVCL_A9TW M 20 643 X 11,12 11 9,12 9,11 14,16 6,9.3 11,12 11,12 3
41 m 74 Frontal rt GSC#171 CVCL_A9TX M 10 437 X,Y 12,13 11,13 8,10 9,12 16,17 8,9.3 10,11 10,11
42 f 77 Parietal pt GSC#172 CVCL_A9TY UM 20 443 X 11 9 9,12 11,13 14,16 7 9,10 11,12 1 , 2 , 3
43 f 64 Occipital rt GSC#181 CVCL_A9TZ M 10 438 X 11,12 12 11,12 8,9 17,18 9 8 10
44 f 53 Frontal pt GSC#184 CVCL_A9U0 UM 25 441 X 11 12,13 9,10 11 17,18 6,9 8 10,12 3
45 m 70 Temporal pt GSC#188 CVCL_A9U1 UM 20 439 X,Y 12,13 12,14 10 9,13 18,19 9.3 8,11 11,13 3
46 m 73 Frontal pt GSC#191 CVCL_A9U2 M 30 444 X,Y 11,13 12 10,11 11 16,17 6,9.3 10,11 11 3
47 m 70 Parietal pt GSC#195 CVCL_A9U3 UM 40 431 X,Y 12 8,11 7,13 12 14,17 9,9.3 8 11 3
47 m 70 Parietal pt GSC#195V CVCL_A9U4 UM 40 447 X,Y 12 8,11 7,13 12 14,17 9,9.3 8 11
48 f 71 Parietal pt GSC#196 CVCL_A9U5 M 25 775 X 12,13 11,12 10,11 10,11 17 7,9 10, 11 11 1 , 2 , 3
49 f 80 Parietal pt GSC#204 CVCL_A9U6 M 25 473 X 10,12 12 11,12 9,12 17,18 6,9.3 8,9 9,11 1 , 2 , 3
50 m 76 Temporal pt GSC#206 CVCL_A9U7 M 50 474 X 10,13 11,13 9,12 11,12 16 9.3 8,10 9,12 3
51 m 68 Temporal rt GSC#208 CVCL_A9U8 UM 40 475 X,Y 11 11 10,11 12,13 15,16 9,9.3 8 11 1 , 2 , 3
52 f 43 Occipital pt GSC#209 CVCL_A9U9 M 40 507 X 11,13 11 10,13 12 13,16 9 11 10,11 1 , 2 , 3
53 m 53 Parietal pt GSC#210 CVCL_A9UA UM 40 476 X,Y 12 11,12 8,12 11,12 16,18 8,9 8,11 9 1 , 2 , 3
54 m 49 Frontal rt GSC#213 CVCL_A9UB M 30 477 X,Y 10,11 8,12 8,11 10,11 17,18 6,8 8 11,12 1 , 2 , 3
55 m 62 Frontal pt GSC#220C CVCL_A9UC UM 30 478 X,Y 11,13 11,13 8,11 10,11 14,17 7,9 8 12,13 1 , 2 , 3
56 m 77 Temporal pt GSC#221 CVCL_A9UD UM 40 479 X,Y 11,13 8,11 8,13 11,12 16,17 9,9.3 9,11 10 1 , 2 , 3
57 m 64 Temporal pt GSC#242 CVCL_A9UE M 25 480 X,Y 12 11 10,14 9,11 17,18 8 8,11 11,12 1 , 2 , 3
58 m 57 Multicentric pt GSC#257 CVCL_A9UF M 50 481 X,Y 12,14 12 9,10 11,12 14,18 8,9.3 8,11 12,13 1 , 2 , 3
59 m 38 Temporal pt GSC#262 CVCL_A9UG UM 35 482 X,Y 11 11 10,11 12,13 16,18 6,9 8 12 1 , 2 , 3
60 m 58 Occipital pt GSC#275 CVCL_A9UH M 50 483 X,Y 10,12 10,12 8,12 9,13 16 6,10 8 10,11 1 , 2 , 3
60 m 58 Occipital rt GSC#275bis CVCL_A9UI M 50 484 X,Y 10,12 10,12 8,12 9,13 16 6,10 8 10,11
61 m 56 Temporal pt GSC#277 CVCL_A9UJ UM NA 508 X,Y 12 11 8,10 12,13 16,17 6,8 8,11 10,12 3
62 m 60 Frontal pt GSC#284 CVCL_A9UK UM NA 485 X,Y 11 11 10 9,11 14 8,9 8 11,13 1 , 2
63 m 50 Cerebellar pt GSC#290 CVCL_A9UL UM NA 486 X,Y 11 11 9,13 10,13 15,19 7,9 8,9 11,12
64 f 61 Frontal pt GSC#291 CVCL_A9UM M 40 487 X 12 8,14 10,14 11,12 17,19 7,9 8,10 12
65 m 47 Frontal pt GSC#298 CVCL_A9UN UM 30 488 X,Y 11,13 9,11 7,11 11,12 15,17 8,9 8 10,11
66 m 65 Temporal pt GSC#309S CVCL_A9UP M 5 489 X 11,13 8,12 9,12 14 16 9,9.3 8 11,12
67 f 54 Parietal pt GSC#314C CVCL_A9UQ M 40 491 X 12,13 11,14 10,12 11 16,19 6,9.3 8,11 10
67 f 54 Parietal pt GSC#314P CVCL_A9UR M 40 490 X 12,13 11,14 10,12 11 16,19 6,9.3 8,11 10
68 m 53 Frontal pt GSC#315 CVCL_A9US UM 50 629 X,Y 9 9,12 10,12 12 16,18 9,9.3 9,10 10
69 m 69 Temporal pt GSC#318 CVCL_A9UT M NA 509 X,Y 12 11,12 9,10 11,12 16,19 8,9.3 9,11 10,12
70 f 51 Parietal pt GSC#323 CVCL_A9UU NA 40 644 X 11,12 11,12 11 11,13 18 8,9.3 11 12,13
71 m 70 Temporal pt GSC#326 CVCL_A9UV UM 45 645 X,Y 9,11 10,12 8,12 11,12 16,17 7,9.3 8,9 12
72 m 82 Frontal pt GSC#327 CVCL_A9UW UM 60 630 X,Y 13 11 8,10 11 17,18 9,9.3 8 10,11
73 m 73 Occipital pt GSC#329 CVCL_A9UX M 30 510 X,Y 11,13 8,14 10,11 9,10 17,19 6 8 11,12
74 m 70 Temporal pt GSC#352 CVCL_A9UY UM 40 657 X,Y 11,13 12,13 9,11 8,11 16,17 6,7 9 11
75 m 61 Frontal pt GSC#361 CVCL_A9UZ UM 50 658 X,Y 11,12 8,12 10 13 14,17 6,7 8,9 10,12
76 m 54 Temporal pt GSC#365 CVCL_A9V0 UM 50 656 X,Y 9,12 11 8,10 10,11 18 6,8 8 11
77 m 43 Temporal pt GSC#366 CVCL_A9V1 UM 35 652 X,Y 11 11,12 8,9 10,13 16,17 9 8,11 12
78 f 65 Frontal pt GSC#369 CVCL_A9V2 M 40 660 X 13 11,12 10 11,12 15 6,9.3 8 10
79 m 69 Frontal pt GSC#381 CVCL_A9V3 M 30 653 X 11,13 11 10,11 11,13 14,17 9 8 11
80 f 54 Temporal pt GSC#384 CVCL_A9V4 M 50 632 X 11,12 12 10,11 10 16,17 7,9 8,11 10,12
81 m 72 Occipital pt GSC#389 CVCL_A9V5 UM 30 655 X,Y 12 8,11 9,10 11,12 14,17 6 8 10,11
82 f 62 Temporal pt GSC#391 CVCL_A9V6 M 15 723 X 10,13 12 8,12 12 16 9,9.3 9,10 9,10
83 m 70 Temporal pt GSC#393 CVCL_A9V7 UM 25 724 X 11 8,14 10 12,13 17,18 9,10 8 10
84 m 64 Frontal pt GSC#394 CVCL_A9V8 M 30 665 X 13 12,13 10,11 11,13 16,17 9.3,10 9,11 11,12
84 m 64 Frontal rt GSC#394bis CVCL_A9V9 M 10 670 X,Y 13 12 10,11 11,13 16,17 9.3,10 9,11 11,12
85 m 49 Temporal pt GSC#395 CVCL_A9VA M 25 627 X 9,11 12 8,11 13 16,17 6,9.3 8,11 10,11
86 f 62 Parietal pt GSC#397 CVCL_A9VB NA 25 654 X 9,11 11,12 9,11 11 15,18 9.3 8,11 10,11
87 m 67 Temporal pt GSC#399 CVCL_A9VC NA 20 623 X,Y 12 12 8,12 11,12 16,17 7,9.3 11 11,12
88 m 71 Occipital pt GSC#401 CVCL_A9VD M 25 633 X,Y 11,12 13 10,11 11,13 15 6,9.3 8 11,12
89 f 68 Frontal pt GSC#403 CVCL_A9VE M 25 624 X 9,11 11,12 12 11 14,15 7,9.3 9,11 12,13
90 m 66 Temporal pt GSC#406 CVCL_A9VF M 20 628 X,Y 12,13 14 10,12 9,12 15,16 9.3 8,9 10
91 f 70 Frontal pt GSC#407 CVCL_A9VG NA 20 651 X 12 9 10,11 11,13 19 9 11 10,11
92 m 69 Temporal pt GSC#411 CVCL_A9VH M 20 646 X 8,11 9,11 8,11 11,12 16,19 9.3 8 10,12
93 f 62 Parietal pt GSC#413 CVCL_A9VI NA 20 625 X 12,13 12 10 11,14 16,17 8,9.3 8,11 10
94 m 71 Frontal pt GSC#415 CVCL_A9VJ UM 15 634 X,Y 12 11,13 8 8,11 18 9,10 8,10 11,12
95 f 85 Frontal pt GSC#416 CVCL_A9VK NA 20 648 X 10 11 8,10 12,13 14,18 9,9.3 8,11 10
96 f 52 Frontal pt GSC#420 CVCL_A9VL M 25 635 X 11,13 12 8,11 12 15,17 6,7 10 11
97 m — Frontal pt GSC#421 CVCL_A9VM UM 20 663 X,Y 10,12 11 9,11 10,12 16,17 9 8,12 10,12
98 f 70 Temporal pt GSC#426 CVCL_A9VN UM 25 668 X 12,13 8,12 10,(13) 9 18 6,9 11 10,11
99 f — Temporal pt GSC#429 CVCL_A9VP UM 20 722 X 13,15 12,13 9,10 9,12 15,18 8 8,11 9,11
100 m 53 Occipital pt GSC#431 CVCL_A9VQ M 40 671 X,Y 13 11 8,11 8,10 17,18 6 9,10 10
101 m 67 Parietal pt GSC#432 CVCL_A9VR M NA 700 X,Y 12,13 8,12 12,13 11,12 16 8 8 10,12
102 f 57 Temporal pt GSC#433 CVCL_A9VS M 20 666 X 11,13 10,12 9,11 11 16,18 6,9 8,11 12,13
103 m 72 Parietal pt GSC#440 CVCL_A9VT M 25 664 X,Y 11,12 9,10 8,9 9,11 15,17 6,9 8,11 9,11
104 f 67 Frontal pt GSC#441 CVCL_A9VU UM 30 692 X 12,13 12 10,11 11 14,17 7,9.3 8,12 10
105 f 51 Frontal pt GSC#442 CVCL_A9VV UM 30 667 X 11 12 7,10 8,12 17 9.3 8 12
106 m 54 Parietal pt GSC#445 CVCL_A9VW UM 30 701 X,Y 10,12 10,11 11 9,12 17,18 7,8 8 10,11
107 m 52 Frontal pt GSC#447P CVCL_A9VX M 25 702 X 12,13 11 11,12 11,12 17,18 6,9 8 11,12
108 m 64 Temporal pt GSC#448 CVCL_A9VY M 20 669 X,Y 11 12 8,13 11,12 16,17 9 8 10,12
109 m 75 Temporal pt GSC#449 CVCL_A9VZ M 20 703 X,Y 11 11 8,11 13, 14 15,17 8,9.3 8,12 10
110 m 76 Temporal pt GSC#450 CVCL_A9W0 M 20 694 X 11,12 11,13 9,11 11,13 17,18 8,9 8 10
111 f 67 Temporal pt GSC#452C CVCL_A9W1 M 20 704 X 12,13 8,12 8 11 17 6,9.3 8 10,12
111 f 67 Temporal pt GSC#452P CVCL_A9W2 M 20 705 X 12,13 8,12 8 11 17 6,9.3 8 10,12
112 m 77 Temporal pt GSC#454 CVCL_A9W3 M 25 706 X 10,11 11 11, 12 11,12 16,17 9,9.3 8,9 12
113 m 74 Temporal pt GSC#455 CVCL_A9W4 UM 25 707 X,Y 12 8 11 12 16,20 6,9.3 11 10
Open in a new tab

Notes: Shown are STR profiles generated in our study from GSCs. Tumor location, primary/recurrent tumor status of glioblastoma tumors that originated GSCs, MGMT gene methylation status and tumor proliferation index (Ki67%) of GSCs are also shown. Previous studies involving GSCs are indicated by the number of reference in the last column of the table.

Abbreviations: f, female; GSC, glioblastoma stem‐like cell; Ki67%, % cells positive to Ki67 proliferation antigen; m, male; M, methylated; MGMT, O6‐methylguanine DNA methyltransferase; NA, not analyzed; pt, primary tumor; rt, recurrent tumor; STR, short tandem repeat; UM, unmethylated.

For GSCs, the MSI status was not analyzed since its frequency, as determined through the amplification of the monucleotide loci, is rare in glioblastoma tumors. Single loci MSI are observed in a low percentage of glioblastoma samples and the presence of high MSI is not a typical feature of this tumor. 18

GSCs were also analyzed for the expression of the markers CD133 and Sox2 (Table S5). Expression of CD133 (n = 45) and Sox2 (n = 43) was 25.0% ± 5.0% (mean ± SEM; range 0%‐95.9%) and 66.1% ± 4.8% (mean ± SEM; range 0.3%‐97.3%), respectively. Our data show that GSCs display different levels of CD133 and Sox2 expression, regardless of their stemness properties. The extensive intra‐ and intertumor heterogeneity of glioblastoma, 19 may account for the inability of surface markers CD133 and Sox2 to characterize GSCs.

3.2. Identification of STR profiles

Eighteen CTSCs and 103 GSCs were genetically identified using STR profiling with two different kits using 10 or 16 different loci (Supplementary Material and Methods). To protect the identity of the subjects, eight core STR loci plus Amelogenin were used to report on the identity of a given sample. The following core loci were recommended, D5S818, D13S317, D7S820, D16S539, vWA, TH01, TPOX and CSF1PO. 20 , 21 The information about the other loci can be shared in strict confidence to researchers and biorepositories. 22 , 23 Tables 1 and 2 show STR profiles of CTSCs and GSCs, respectively. As an example, the STR profile for GSC#447P cell line (STR ID 702) is given in Figure S1.

Amelogenin marker was used for gender determination. 24 This marker is located on the gonosomal chromosomes, in Xp22.1‐22.3 (AMELX) and Yp11.2 (AMELY). Its DNA fragments, obtained in PCR using specific primers for intron 3, differ by 6 bp, between X and Y chromosomes, because the AMELX contains a 6 bp deletion in intron 3 (CRCh38.12, 11,296,918 and 11,296,919, GenBank).

Failure in amelogenin sex gene detection is rare in healthy individuals 25 and in diploid cells. However, AMELY chromosomal losses are highly frequent in tumor cell lines, hence exclusive AMELX is not predictive for the authentication in samples originated from male patients. 12 Cell lines derived from tumor samples can lose part of the Y chromosome during culture, therefore sex determination can only be indicative.

Among the GSCs that arose from 66 male individuals, 16 cell lines showed only AMELX (GSC#1, GSC#83 and 83.2, GSC#142, GSC#148, GSC#170, GSC#206, GSC#309S, GSC#381, GSC#393, GSC#394, GSC#395, GSC#411, GSC#447P, GSC#450, GSC#454). Therefore, about 24% of the male‐derived cell line profiles seem to have lost AMELY. It has been reported that about 40% to 45% of cell lines purportedly derived from males lacked the AMELY allele. 26

3.3. Comparison of STR profiles

The analysis of STR profiles of CTSCs and GSCs by CLIMA 13 revealed that some cell lines were derived from the same individual. Among GSCs, the following cell lines derived from the same patient have profile similarity, GSC#23C and GSC#23p (93.75% of similarity); GSC#83, GSC#83.2 (93.75% of similarity); GSC#30P and GSC#30PT (100% of similarity); GSC#195 and GSC#195V (100% of similarity); GSC#314P and GSC#314C (100% of similarity). The following cell lines derived from primary and secondary surgery of the same patient show profile similarity, GSC#275 and GSC#275bis (100% of similarity); GSC#394, GSC#394bis (100% of similarity).

In some cell lines, PBMCs and the primary tumor tissues (T) were also analyzed for STR profiles. Results were as follows, GSC#298 and #298 PBMCs (100% of similarity); GSC#309s, #309T and #309 PBMCs (100% of similarity); 314T and 314 PBMCs (100%); GSC#318 and #318T (100%). All the other cell lines have unique profiles, that do not match with other profiles for a percentage higher than 90%. All unique profiles have been confirmed in CLASTR.

4. DISCUSSION

Human cell lines obtained from CSCs represent an invaluable model for studying the properties of tumors. 27 These cells provide new insights into the biology of tumors and models that use the CSCs are essential tools for translational research. For example, we previously demonstrated that CSC‐enriched spheroid cultures faithfully capture important features of primary colorectal tumors in terms of both genetic landscape and drug sensitivity. 2 , 3 , 4 , 5 , 6 In a recent study, we provided an extensive analysis of CTSC response to EGFR‐targeted therapy in vivo, leading to a deeper understanding of the molecular determinants of therapy resistance and sensitivity to combination therapies. 6 In another study on the translational impact of the CSC model, we demonstrated that resistance of GSCs to standard treatment (ie, radiation therapy and temozolomide) relates to the clinical outcome of donor patients. 3 In addition to demonstrating the clinical relevance of CSCs, these studies suggest how this model may guide therapeutic strategies in terms of both response predictions to current treatment and more appropriate drug selection. 6

In the present study, we detected MSI‐H in 7 out 18 CTSC cell lines (38%) that is higher than expected in CRC. There are two possible explanations for this result. The first is that 13 (72%) tumor samples, which the CTSCs were isolated from, came from right‐sided colon cancers that harbor the MSI‐H phenotype most often. 28 An alternative explanation is that the cultured CRC cell lines show MSI‐H more frequently than the parent tumor, 29 suggesting that MSI‐H tumors can be more easily expanded in vitro. Most of the cell lines described in this article have been used in earlier studies 1 , 2 , 3 , 4 , 5 , 6 , 9 , 10 , 11 , 12 , 13 (see Tables 1 and 2 for details).

Despite the success of using cell lines as models to advance cancer research, misidentification of cell lines is a widespread problem. 7 , 8 , 9 , 12 , 30 , 31 Authentication testing is an effective way to solve the problem, for this reason the disclosure of false or misidentified cell lines is the principal aim of the International Cell Line Authentication Committee (www.ICLAC.org), a voluntary, independent scientific committee, established in 2012. ICLAC produces important guidelines, such as “Guide to Human Cell Line Authentication” and “Obtaining Cell Lines from Reliable Sources.” ICLAC was established after the publication of a consensus Standard for human cell line authentication by STR profiling. 12 Cellosaurus is a cell line knowledge resource containing information about 92 500 human cell lines and reports data about problematic (contaminated/misidentified) cell lines. Recently, a collaboration between the Cellosaurus database 32 and the Resource Identification Initiative (https://f1000research.com/articles/4-134/v2) determined the use of an unique research resource identifier to flag each established cell line for searches and data analysis. 33

Using STR profiling, we generated for each CSC line a unique molecular identity pattern. STR profiles from CTSCs and GSCs were compared both with cell line profiles included in CLIMA 2.1 database 13 and with cell lines of Cellosaurus using the CLASTR search tool. Besides STR profiles, for each CTSC and GSC line, clinical data of the patients are reported such as tumor location, stage and mutations, MGMT methylation status and tumor proliferation index, MSI, expression of mismatch repair proteins (Tables 1 and 2; Tables S2 and S3) and the expression of molecular markers (Tables S4 and S5).

The cell lines used in our study will be available to researchers through Material Transfer Agreement (MTA).

CONFLICT OF INTEREST

The authors declared no potential conflicts of interest.

ETHICS STATEMENT

Glioblastoma tissue samples were harvested from patients undergoing craniotomy at the Institute of Neurosurgery, Università Cattolica del Sacro Cuore (UCSC), Rome, Italy. All the patients provided written informed consent according to the research proposals approved by the Ethical Committee of UCSC. Fresh human colorectal cancer tissues were obtained in accordance with the standards of the ethics committee on human experimentation of the Istituto Superiore di Sanità (authorization no. CE5ISS 09/282). All the patients provided written informed consent. Cell lines obtained from tumor stem‐like cells were de‐identified to protect patient health information.

Supporting information

Appendix S1: Supporting Information

Click here for additional data file. (11.6MB, pdf)

ACKNOWLEDGMENTS

This work was supported by the grants from Italian Ministry of Health (Alliance Against Cancer network ACC to Barbara Parodi) (RF‐2016‐02361089 to Lucia Ricci‐Vitiani) and Associazione Italiana per la Ricerca sul Cancro (AIRC) (IG 2014 n.15584 to Lucia Ricci‐Vitiani and IG 2019 n.23154 to Roberto Pallini).

Visconti P, Parodi F, Parodi B, et al. Short tandem repeat profiling for the authentication of cancer stem‐like cells. Int. J. Cancer. 2021;148:1489–1498. 10.1002/ijc.33370

Paola Visconti and Federica Parodi contributed equally to the study.

Funding information Associazione Italiana per la Ricerca sul Cancro, Grant/Award Numbers: IG 2013 14574, IG 2014 15584; Italian Ministry of Health, Grant/Award Number: RF‐2016‐02361089

DATA AVAILABILITY STATEMENT

The cell lines used in this study will be available to researchers through a Material Transfer Agreement (MTA). The presented STR profiles and clinical information of our CSCs are available in the Cellosaurus database (ExPASy), under the RRID numbers listed in Tables 1 and 2. In addition, the STR profiles are also uploaded to the Cell Line Integrated Molecular Authentication Database 2.1 (CLIMA 2.1) (http://bioinformatics.hsanmartino.it/clima2/). Other data supporting the findings of this study are available from the corresponding author upon request.

REFERENCES

  • 1. Grande S, Palma A, Ricci‐Vitiani L, et al. Metabolic heterogeneity evidenced by MRS among patient‐derived Glioblastoma Multiforme stem‐like cells accounts for cell clustering and different responses to drugs. Stem Cells Int. 2018;2018:3292704. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 2. Marziali G, Buccarelli M, Giuliani A, et al. A three‐microRNA signature identifies two subtypes of glioblastoma patients with differentclinicaloutcomes. Mol Oncol. 2017;11:1115‐1129. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 3. D'Alessandris QG, Biffoni M, Martini M, et al. The clinical value of patient‐derived glioblastoma tumorspheres in predicting treatment response. Neuro Oncol. 2017;19:1097‐1108. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 4. Pallini R, Ricci‐Vitiani L, Banna GL, et al. Cancer stem cell analysis and clinical outcome in patients with glioblastoma multiforme. Clin Cancer Res. 2008;14:8205‐8212. [DOI] [PubMed] [Google Scholar]
  • 5. Ricci‐Vitiani L, Lombardi DG, Pilozzi E, et al. Identification and expansion of human colon‐cancer‐initiating cells. Nature. 2007;445:111‐115. [DOI] [PubMed] [Google Scholar]
  • 6. De Angelis ML, Zeuner A, Policicchio E, et al. Cancer stem cell‐based models of colorectal cancer reveal molecular determinants of therapy resistance. Stem Cells Transl Med. 2016;5:511‐523. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 7. Korch C, Varella‐Garcia M. Tackling the human cell line and tissue misidentification problem is needed for reproducible biomedical research. Adv Molec Pathol. 2018;1:209‐228. [Google Scholar]
  • 8. Capes‐Davis A. Cell line detective work. Basic principles and molecular applications. Adv Molec Pathol. 2018;1:229‐238. [Google Scholar]
  • 9. Schweppe RE, Korch C. Challenges and advances in the development of cell lines and xenografts. Adv Molec Pathol. 2018;1:239‐251. [Google Scholar]
  • 10. Nims RW. Historical and current methods for detecting interspecies and intraspecies cell mixtures and thereby assuring cell line purity. Adv Molec Pathol. 2018;1:253‐264. [Google Scholar]
  • 11. Signore M, Buccarelli M, Pilozzi E, et al. UCN‐01 enhances cytotoxicity of irinotecan in colorectal cancer stem‐like cells by impairing DNA damage response. Oncotarget. 2016;7(28):44113‐44128. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 12. ATCC® Standards Development Organization . Designation: ASN‐0002. Authentication of Human Cell Lines: Standardization of STR Profiling. ANSI/ATCC ASN‐0002‐2011; 2011.
  • 13. Romano P, Manniello A, Aresu O, Armento M, Cesaro M, Parodi B. Cell line data base: structure and recent improvements towards molecular authentication of human cell lines. Nucleic Acids Res. 2009;37(suppl_1):D925‐D932. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 14. Falchetti ML, Pierconti F, Casalbore P, et al. Glioblastoma induces vascular endothelial cells to express telomerase in vitro. Cancer Res. 2003;63(13):3750‐3754. [PubMed] [Google Scholar]
  • 15. Esteller M, Hamilton SR, Burger PC, Baylin SB, Herman JG. Inactivation of the DNA repair gene O6‐methylguanine‐DNA methyltransferase by promoter hypermethylation is a common event in primary human neoplasia. Cancer Res. 1999;59(4):793‐797. [PubMed] [Google Scholar]
  • 16. Robin T, Capes‐Davis A, Bairoch A. CLASTR: the Cellosaurus STR similarity search tool ‐ a precious help for cell line authentication. Int J Cancer. 2019;146:1299‐1306. 10.1002/ijc.32639. [DOI] [PubMed] [Google Scholar]
  • 17. Kazama S, Kishikawa J, Kiyomatsu T, et al. Expression of the stem cell marker CD133 is related to tumor development in colorectal carcinogenesis. Asian J Surg. 2018;41(3):274‐278. [DOI] [PubMed] [Google Scholar]
  • 18. Maxwell JA, Johnson SP, McLendon RE, et al. Mismatch repair deficiency does not mediate clinical resistance to temozolomide in malignant glioma. Clin Cancer Res. 2008;14(15):4859‐4868. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 19. Ludwig K, Kornblum HI. Molecular markers in Glioma. J Neurooncol. 2017;134(3):505‐512. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 20. Stabley DL, Holbrook J, Harris AW, et al. Establishing a reference dataset for the authentication of spinal muscular atrophy cell lines using STR profiling and digital PCR. Neuromuscul Disord. 2017;27:439‐446. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 21. Capes‐Davis A, Reid YA, Kline MC, et al. Match criteria for human cell line authentication: where do we draw the line? Int J Cancer. 2013;132:2510‐2519. [DOI] [PubMed] [Google Scholar]
  • 22. Isasi R, Andrews PW, Baltz JM, et al. Identifiability and privacy in pluripotent stem cell research. Cell Stem Cell. 2014;14:427‐430. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 23. Bredenoord AL, Mostert M, Isasi R, Knoppers BM. Data sharing in stem cell translational science: policy statement by the international stem cell forum ethics working party. Regen Med. 2015;10:857‐861. [DOI] [PubMed] [Google Scholar]
  • 24. Nakahori Y, Takenaka O, Nakagome Y. A human X‐Y homologous region encodes amelogenin. Genomics. 1991;9:264‐269. [DOI] [PubMed] [Google Scholar]
  • 25. Steinlechner M, Berger B, Niederstatter H, Parson W. Rare failures in amelogenin sex test. Int J Legal Med. 2002;116:117‐120. [DOI] [PubMed] [Google Scholar]
  • 26. Liang‐Chu MM, Yu M, Haverty PM, et al. Human biosample authentication using the high‐throughput, cost‐effective SNPtrace(TM) system. PLoS One. 2015;10(2):e0116218. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 27. Baiocchi M, Biffoni M, Ricci‐Vitiani L, Pilozzi E, De Maria R. New models for cancer research: human cancer stem cell xenografts. Review. Curr Opin Pharmacol. 2010;10(4):380‐384. [DOI] [PubMed] [Google Scholar]
  • 28. Raut PC, Pawlik MP, Rodriguez‐Bigas MA. Clinicopathologic features in colorectal cancer patients with microsatellite instability. Mutat Res. 2004;568:275‐282. [DOI] [PubMed] [Google Scholar]
  • 29. Medico E, Russo M, Picco G, et al. The molecular landscape of colorectal cancer cell lines unveils clinically actionable kinase targets. Nat Commun. 2015;6:7002. [DOI] [PubMed] [Google Scholar]
  • 30. Freedman LP, Gibson MC, Ethier SP, Soule HR, Neve RM, Reid YA. Reproducibility: changing the policies and culture of cell line authentication. Nat Methods. 2015;12:493‐497. [DOI] [PubMed] [Google Scholar]
  • 31. Nims RW, Reid Y. Best practices for authenticating cell lines. In Vitro Cell Dev Biol Anim. 2017;53:880‐887. [DOI] [PubMed] [Google Scholar]
  • 32. Bairoch A. The Cellosaurus, a cell line knowledge resource. J Biomol Tech. 2018;29:25‐38. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 33. Capes‐Davis A, Bairoch A, Barrett T, et al. Cell lines as biological models: practical steps for more reliable research. Chem Res Toxicol. 2019;32(9):1733‐1736. [DOI] [PMC free article] [PubMed] [Google Scholar]

Associated Data

This section collects any data citations, data availability statements, or supplementary materials included in this article.

Supplementary Materials

Appendix S1: Supporting Information

Click here for additional data file. (11.6MB, pdf)

Data Availability Statement

The cell lines used in this study will be available to researchers through a Material Transfer Agreement (MTA). The presented STR profiles and clinical information of our CSCs are available in the Cellosaurus database (ExPASy), under the RRID numbers listed in Tables 1 and 2. In addition, the STR profiles are also uploaded to the Cell Line Integrated Molecular Authentication Database 2.1 (CLIMA 2.1) (http://bioinformatics.hsanmartino.it/clima2/). Other data supporting the findings of this study are available from the corresponding author upon request.

Articles from International Journal of Cancer are provided here courtesy of Wiley

RESOURCES