Organoid co-culture model of the cycling human endometrium in a fully synthetic extracellular matrix enables the study of epithelial-stromal crosstalk

. Author manuscript; available in PMC: 2024 Aug 11.

Published in final edited form as: Med. 2023 Aug 11;4(8):554–579.e9. doi: 10.1016/j.medj.2023.07.004

Key resources table

REAGENT or RESOURCE	SOURCE	IDENTIFIER
Antibodies
Vimentin	abcam	ab8978
DAPI	ThermoFischer	R37606
AlexaFluor^™ 488 phalloidin	ThermoFischer	R37110
EpCAM	Abcam	ab7504
Laminin (LMN)	Abcam	ab11575
Cleaved Caspase-3	Abcam	ab2302
Progesterone receptor (PGR)	Abcam	ab16661
Progestagen associated endometrial protein (PAEP)	Abcam	ab270454
LiveDead Viability Kit	Invitrogen	R37601
acetylated 𝛼-Tubulin- AF594	Abcam	ab195889
Ki67	Abcam	ab15580
Vimentin-AF594	Abcam	ab154207
Vimentin-AF647	Abcam	ab195878
Goat pAb to Rb IgG-AF594	Abcam	ab150080
Goat pAb to Ms IgG-AF647	Abcam	ab150115
smooth muscle actin (SMA)	Invitrogen	MA-37027
E-cad	R&D	AF748
ESRalpha (ESR1)	Abcam	ab16660
EpCAM	Abcam	ab218448

Bacterial and virus strains
N/A
Biological samples
Human uterine tissues	This paper	Protocol number IRB-P001994
Chemicals, peptides, and recombinant proteins
Mifepristone (RU-486)	Sigma	M8046
17-β estradiol	Sigma	50-28-2
Medroxyprogesterone acetate (MPA)	Sigma	71-58-9
PEG-20 (8-arm 20kDa PEG-VS)	Jenkem	8ARM(TP)-VS
XL-IA	(Ac)-GCRD-LPRTG-GPQGIAGQ-DRCG-(Am)	Custom peptide Boston Open Labs (Cambridge, MA), GenScript (Piscataway, NJ), or CPC Scientific (Sunnyvale, CA).
PHSRN-K-RGD	(Ac)-PHSRNGGGK-(GGG-ERCG-(Am))-GGRGDSPY-(Am)	Custom peptide Boston Open Labs (Cambridge, MA), GenScript (Piscataway, NJ), or CPC Scientific (Sunnyvale, CA).
GFOGER	‘‘(Ac)-GGYGGGPG(GPP)₅GFOGER(GPP)₅GPC-(Am)[45,46]	Custom peptide Boston Open Labs (Cambridge, MA), GenScript (Piscataway, NJ), or CPC Scientific (Sunnyvale, CA).
FN-binder	(Am)-GCRE-TLQPVYEYMVGV-(Ac)	Custom Peptide Boston Open Labs (Cambridge, MA), GenScript (Piscataway, NJ), or CPC Scientific (Sunnyvale, CA).
Critical commercial assays
Click-iT^™ EdU Cell Proliferation Kit	Thermo Fisher	C10424-647
Directzol RNA Mini-Prep kit	Zymo Research	R2051
High-Capacity RNA-to-cDNA Kit	ThermoFisher Scientific	4387406
PureLink DNase Set	Thermofisher Scientific	12185010
Prolactin ELISA duoset	R&D Systems/Fisher	DY682
Matrigel Phenol free	Fisher/Corning	356231
TMT10plex kit	Pierce	90110
barcoded mRNA capture beads	ChemGenes	N/A
Human XL Cytokine Luminex Performance Panel Premixed Kit	R&D Systems	LKTM014
Luminex Performance Human MMP Magnetic Panel	R&D Systems	LMPM000
Maxima H Minus Reverse Transcriptase	ThermoFisher	EP0751
KAPA Hifi PCR Mastermix	Kappa Biopsystems	KR0368

Deposited data
Bulk RNAseq data and single cell RNA sequencing data	This paper, and Baugh and Goods et al.⁴⁴	https://fairdomhub.org/studies/1139; dbGAP Accession number is phs003326.v1.p1.
Raw imaging data	This paper.	https://fairdomhub.org/studies/1139
Proteomics	Baugh and Goods et al.⁴⁴	https://fairdomhub.org/studies/1139
Experimental models: Cell lines
Human: Primary endometrial epithelial cells (EEOs)	This paper	Protocol number IRB-P001994
Human: Primary endometrial stromal cells (ESCs)	This paper	Protocol number IRB-P001994
Experimental models: Organisms/strains
N/A
Oligonucleotides
GAPDH	Hs02786624_g1	157
PGR	Hs01556702_m1	77
ESR1	Hs01046816_m1	65
PAEP	Hs01046123_g1	66
ITGA2	Hs00158127_m1	67
ITGA5	Hs01547673_m1	54
ITGB4	Hs00236216_m1	65
ITGA6	Hs01041011_m1	64
ITGA3	Hs01076879_m1	85
ITGA1	Hs00235006_m1	87
ITGB2	Hs00164957_m1	76
ITGAV	Hs00233808_m1	64
ITGA8	Hs00233321_m1	89
ITGB1	Hs01127536_m1	74
Recombinant DNA
N/A
Software and algorithms
ImageJ	Rasband, W.S., ImageJ, U. S. National Institutes of Health, Bethesda, Maryland, USA, https://imagej.nih.gov/ij/, 1997-2018.	https://imagej.nih.gov/ij/
Imaris (RRID:SCR_007370)	Imaris 9.7 (Bitplane)	http://www.bitplane.com/imaris/imaris
GraphPad Prism (RRID:SCR_002798)	Graph pad Prism 8	https://www.graphpad.com/
Other
N/A