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Organoid Culture

Organoids are in-vitro 3D cell aggregates derived from stem cells which are not only capable of self-organization and long term self-renewal, but which also exhibit similar function to the tissues from which they were derived.1-4) This is achieved through the use of physical and biochemical cues which are able to recapitulate cells’ natural environment within living tissue. As such, organoids are able to overcome many of the limitations of existing culture models (2D monolayers, 3D aggregates (such as spheroids), animal models, etc.). Current applications for organoid culture systems include those in:

  • Developmental Biology5-9)
  • Disease Pathology10-22)
  • Drug Toxicity / Efficacy Testing23-26)
  • Regenerative Medicine27-30)
  • Personalized Medicine15,18,26,31,32)

Organoids can be generated by imbedding either primary tissue (human adult stem cells (hAdSCs)) or pluripotent stem cells (induced pluripotent stem cells (iPSCs) or embryonic stem cells (ESCs)) in the appropriate matrix components and applying appropriate signaling molecules / proteins.3)

Some examples of organoid culture conditions can be found below (matrix components, base culture medium, and chemically defined supplements not shown).

Tissue Type Origin Culture Conditions Reference
Intestine hAdSCs A83-01 [A3324], Nicotinamide [N0078], N-Acetylcysteine [A0905], Y-27623 [Y0018], SB202190, EGF, Rspondin1, Noggin, Wnt-3a 33
hPSCs EGF, Rspondin1, FGF-4, Noggin, Wnt-3a, Activin A 34
Colon hAdSCs A83-01 [A3324], Nicotinamide [N0078], N-Acetylcysteine [A0905], Y-27623 [Y0018], SB202190, EGF, Rspondin1, Noggin, Wnt-3a 33
Stomach hAdSCs A83-01 [A3324], Nicotinamide [N0078], N-Acetylcysteine [A0905], Y-27623 [Y0018], EGF, Rspondin, FGF-10, Gastrin, Noggin, Wnt-3a 11
hPSCs CHIR 99021 [C2943], Y-27623 [Y0018], Retinoic Acid [R0064], EGF, FGF-4, BMP-4, Noggin, Activin A, Wnt-3a 12
Lung hAdSCs A83-01 [A3324], Nicotinamide [N0078], Y-27632 [Y0018], SB202190, Rspondin1, FGF-7, FGF-10, Noggin 35
hPSCs hPSC Differentiation:
SB431542 [B4003], SANT-2, SU-5402, bFGF, Noggin, SHH, SAG, Activin A

Organoid Culture:
CHIR 99021 [C2943], SB431542, FGF-4, Noggin
36
Brain hPSCs Y-27632 [Y0018], Heparin [H0393], 2-Mercaptoethanol [M0058], bFGF, Insulin 37
Liver hAdSCs Organoid Culture:
Y-27632 [Y0018], A83-01 [A3324], Nicotinamide [N0078], N-Acetylcysteine [A0905], Forskolin [F0855], EGF, Rspondin1, FGF-10, Gastrin, Noggin, Wnt-3a, HGF

Hepatocyte Differentiation:
A83-01 [A3324], DAPT [D4257], Dexamethasone [D1961], EGF, FGF-19, BMP-7, Gastrin, HGF
17
hPSCs bFGF, BMP-4, Activin A, HGF, Oncostatin M 38
Pancreas hAdSCs A83-01 [A3324], Nicotinamide [N0078], N-Acetylcysteine [A0905], EGF, Rspondin1, FGF-10, Gastrin, Noggin, Wnt-3a 20
Kidney hPSCs Differentiation:
CHIR 99021 [C2943], 2-Mercaptoethanol [M0058], Heparin [H0393], Retinoic Acid [R0064], 1-Thioglycerol [T0905], bFGF, FGF-9, BMP-2, BMP-4, Insulin, Activin A, Holo-transferrin

Organoid Culture:
CHIR 99021 [C2943], Heparin [H0393], FGF-9, HGF, GDNF
39, 40
Prostate hAdSCs Y-27623 [Y0018], A83-01 [A3324], Nicotinamide [N0078], N-Acetylcysteine [A0905], SB202190, Prostaglandin E2 [P1884], Testosterone [T0027], EGF, Rspondin1, FGF-10, bFGF, Noggin 41
hPSCs Prostate Differentiation:
FGF-10, Activin A, Wnt-10b

Organoid Culture:
Retinoic Acid [R0064], Testosterone [T0027], EGF, Rspondin1, Noggin
42

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