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Table 2 Summary of sources and culture conditions used in the development of various reproductive organoids

From: Organoid technology in female reproductive biomedicine

Organoids Source Culture conditions Cell types in organoids Matrix Generation efficiency Reference
Ovarian cancer Ovarian cancer tissue Establishment: Advanced DMEM/F12, Glutamax, HEPES, Noggin, Rspo1, WNT3A, B27, FGF10, bFGF, nicotinamide, N-acetyl-L-cysteine, A83–01, Heregulinβ-1, hEGF, IGF-1, HGF, Forskolin, Hydrocortisone, NRG1, p38i (SB203580), β-Estradiol, Y-27632
Test components:
carboplatin, paclitaxel, alpelisib, pictilisib, MK2206, AZD8055, Niraparib, adavosertib, gemcitabine, doxorubicin, nutlin-3
Disease and original tumor phenotype Cultrex
growth factor reduced BME type 2
33–65% [15, 16]
Fallopian tube human iPSC lines (87iCTR-n3, 01iMEC-n4, 14iCTR-n6) Mesoderm induction: Activin A, CHIR99021, Y27632
Intermediate mesoderm induction: BMP4, CHIR99021, Y27632
The fallopian tube epithelial cells differentiation: WNT4, Follistatin
Establishment:
DMEM/F12, reconstituted Ultroser, Y-27632, estrogen, progesterone,
conditioned media from FTE cells
Maturation: cultured in 3D Matrigel for an extended period with estrogen and progesterone supplemented media
Ciliated (TUBB4A and FOXJ1) cells
secretory (PAX8) cells
Matrigel NR [22]
  Lin − EPCAM+ FTE cells Establishment: Advanced DMEM/F12, GlutaMAX, B27, EGF, TGFBR1 Kinase Inhibitor IV, Y-27632
Differentiation: RSPO1
PAX8+ secretory cells
acetylated tubulin (AcTUB) + ciliated cells
Matrigel NR [23]
  Epithelial progenitor (EpCAM+) cell Establishment: Advanced DMEM/F12, conditioned human Wnt3A medium, conditioned human RSPO1 medium, GlutaMAX, B27, N2, human EGF, human noggin, human FGF10, nicotinamide, Y-27632, TGF-β R Kinase Inhibitor IV
hormonal stimulation: beta-oestradiol, progesterone
Pax8-positive secretory cells
Pax8 negative, acetylated tubulin-positive ciliated cells
Matrigel NR [25]
Endometrium the mouse endometrial glandular-type fragments Establishment: DMEM/F12, GlutaMAX, B27, ITS, FGF10, nicotinamide, WNT3A; R-spondin 1, Noggin, A83–01
Hormonal stimulation: beta-oestradiol, progesterone
secretory cells
ciliated cells
Matrigel NR [4]
  the human endometrial glandular-type fragments Establishment: WNT3A; R-spondin 1, EGF, FGF10, Noggin, A83–01, ITS, N-acetyl-L-cysteine, p38 inhibitor SB202190Hormonal stimulation: beta-oestradiol, progesterone secretory cells
ciliated cells
Matrigel NR [4]
  Dissociated endometrial cancer cells Establishment: DMEM/F12, B27, Glutamax, N-acetyl cysteine, Primocin, nicotinamide, A 83–01, SB 202190, Y- 27632, 17-A estradiol
Test components:
Megestrol acetate, fulvestrant, letrozole, mifepristone, erlotinib, linsitinib; Selleckchem, BGJ-398, BBI608, cisplatin, paclitaxel.
Epithelial cells
mesenchymal derivatives
growth factor reduced BME type 2 NR [26]
  Endometrial epithelial cells Establishment: ExM: Advanced DMEM/F12, N2, B27 minus vitamin A, Primocin, N-Acetyl-L-cysteine, L-glutamine, Recombinant human EGF, Recombinant human Noggin, Recombinant human Rspondin-1, Recombinant human FGF-10, Recombinant human HGF, ALK-4, −5, −7 inhibitor A83–01, Nicotinamide
Differentiation: β-oestradiol, progesterone, cAMP, prolactin, human chorionic gonadotropin, human placental lactogen
Hormonal stimulation: β-oestradiol, progesterone, cAMP
secretory (PAEP+) cells
ciliated (acetylated-α-tubulin+) cells
Matrigel 100% [3]
  Endometriotic epithelial cells (ECT-O)
Epithelial cell from hyperplastic endometrium (HYP-O)
Establishment; SOM; DMEM/F12 + L-glutamine and Hepes, RSPO1-conditioned medium, Noggin, B27, N2, Glutamax, Insulin Transferrin Selenium, Nicotinamide, A83–01, N-acetyl L-cysteine, EGF, bFGF, FGF-10, SB202190 (p38i) secretory cells
ciliated cells
Matrigel ECT-O; 60%
HYP-O; 70%
[24]
  Epithelial cell from endometrial cancer Establishment; SOM; DMEM/F12 + L-glutamine and Hepes, RSPO1-conditioned medium, Noggin, B27, N2, Glutamax, Insulin Transferrin Selenium, Nicotinamide, A83–01, N-acetyl L-cysteine, EGF, bFGF, FGF-10, insulin-like growth factor 1 (IGF1), hepatocyte growth factor (HGF) and lipids
Test components:
paclitaxel, 5-fluorouracil, carboplatin, doxorubicin and everolimus
NR Matrigel EC-O; 40%; [24]
Trophoblast villous cytotrophoblasts (vCTBs), purified from pooled first-trimester placental tissues Establishment: basic trophoblast organoid medium (b-TOM): advanced DMEM/F12, HEPES, B27, N2, glutamine, R-spondin, A83–01, rhEGF, rmHGF, prostaglandin E2, CHIR99021, Noggin, EGF, R-spondin, CHIR99021, A83–01
Differentiation:
lacking R-spondin and CHIR99021, inhibitor of Wnt response-1 (IWR-1)
cytotrophoblasts (CTB) syncytiotrophoblasts (STB)
extravillous trophoblast (EVT)
Matrigel 100% [5]
  trophoblast-enriched cell suspensions Establishment:
basal trophoblast organoid medium (TOM): EGF, FGF2, CHIR99021, A83–01, R-spondin 1, HGF, PGE2, Y-27632, nicotinamideDifferentiation:
EVT differentiation medium (EVTM): advanced DMEM/ F12, 2-mercaptoethanol, BSA, ITS-X, NRG1, A83–01, KSR.
Typically after days 7–10, the medium was changed to EVTM without NRG1 for a further 7–10 days.
syncytiotrophoblast (SCT)
villous cytotrophoblast (VCT)
HLA-G+ extravillous trophoblast cells (EVT) cells
Matrigel 91% [27]
Cervical organoid cervical clear cell carcinoma cells Establishment:
DMEM/F12, human EGF, R-spondin1, Noggin, Y27632, Jagged-1, l-glutamine
Test components:
paclitaxel, cisplatin, gemcitabine hydrochloride, crizotinib, and SU11274
Atypical
cells with clear cytoplasm concordant with morphological features of the original tumor
Matrigel   [28]
  1. Abbreviations: A.83–01 Selective inhibitor of ALK4,5,7, EGF Epidermal growth factor, FGF Fibroblast growth factor, HGF Hepatocyte growth factor, NRG1 Neuregulin-1, Rho-KI Rho kinase inhibitor, RSPO1 R-spondin-1, NR Not-reported