Figure - PMC

Skip to main content

An official website of the United States government

Here's how you know

Here's how you know

Official websites use .gov
A .gov website belongs to an official government organization in the United States.

Secure .gov websites use HTTPS
A lock ( ) or https:// means you've safely connected to the .gov website. Share sensitive information only on official, secure websites.

View full-text article in PMC

. 2022 Oct 17;2(11):100317. doi: 10.1016/j.crmeth.2022.100317

Search in PMC
Search in PubMed
View in NLM Catalog
Add to search

© 2022 The Authors

This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).

PMC Copyright notice

Reprogramming HDFs and PBMCs into naive human iPSCs with a fast SeV vector removal system and modified SeV-KLF4 vectors

(A) qPCR analysis of SeV genome expression in naive iPSCs reprogrammed with the CytoTune 2.0 or 2.0L SeV reprogramming kit using TaqMan probes. Values are normalized to GAPDH expression and shown as the mean ± SD. n = 3 for each point.

(B) Schematic structure of the SeV vectors and the modified reprogramming cocktail. We changed the SeV18 + KLF4/TSΔF (KLF4/TS) vector in the CytoTune-2.0 or 2.0L reprogramming kit to one of three modified vectors: SeV18 + KLF4/TS12ΔF (KLF4/TS12), SeV18 + KLF4/PmiR367T2/TSΔF (KLF4/miR/TS), or SeV18 + KLF4/PmiR367T2/TS12ΔF (KLF4/miR/TS12). KLF4/miR/TS12 was developed for this study.

(C) Experimental design for the derivation of naive human iPSCs from HDFs or PBMCs using the modified SeV-OSKL cocktail and incubation temperature shift after the generation of naive human iPSCs.

(D) Number of naive human iPSC colonies generated from HDFs at day 14. Each of the SeV-KLF4/TS vectors was co-infected with SeV-KOS and SeV-LMYC. Data are shown as the mean ± SD. n = 3. ∗p < 0.05.

(E) qPCR analysis of SeV genome expression in naive iPSCs derived from HDFs reprogrammed with SeV-KOS, SeV-LMYC, and the respective SeV-KLF4 vectors using TaqMan probes. Data are shown as the mean ± SD. n = 3 of each point.

(F) qPCR analysis of SeV genome expression in HDF-derived naive iPSCs (nOSKL_1, 2) and PBMC-derived naive iPSCs (nOSKL_3, 4) reprogrammed with SeV-KOS, SeV-LMYC, and SeV-KLF4/miR/TS12 vectors at days 14 and 34 using TaqMan probes. Data are shown as the mean ± SD. n = 3.

(G) Representative phase-contrast images of naive iPSCs reprogrammed with SeV-KOS, SeV-LMYC, and SeV-KLF4/miR/TS12 vectors on iMEF feeder cells or in feeder-free conditions. Scale bars, 200 μm.