Table 1. Frequently used growth media constituents, their working mechanisms and effects, as well as applications
Growth medium constituentsWorking mechanism in ISCsEffect on ISCs and application
WNT3aaActivates canonical WNT signaling (Clevers & Nusse, 2012)Stimulates crypt cells proliferation and maintains the stem cell state (Clevers & Nusse, 2012; Farin et al, 2012; Krausova & Korinek, 2014)
R‐spondin 1aAugments WNT/β‐catenin signaling (de Lau et al, 2014)Stimulates crypt cell proliferation and maintains stem cell state (Farin et al, 2012; Krausova & Korinek, 2014; de Lau et al, 2014)
CHIR99021Stimulates canonical WNT signaling (Yin et al, 2014)Stimulates stem cell proliferation and can be used in combination with VPA, when growing single mouse ISCs in absence of Paneth cells (Yin et al, 2014)
Valproic acidInhibits histone deacetylase and activates Notch signaling (Yin et al, 2014)Maintains proliferative crypts and blocks secretory differentiation (Sato et al, 2011b). Can be used in combination with CHIR99021 when growing single mouse ISCs in absence of Paneth cells (Yin et al, 2014)
NogginaInhibits BMP signaling (Haramis et al, 2004)Stimulates crypt formation (Haramis et al, 2004)
Jagged‐1Activates Notch signaling (Sato et al, 2009)

Maintains the stem cell state, and promotes proliferation, while blocking secretory differentiation, thereby maintaining proliferative crypts (Stanger et al, 2005; Van Dussen et al, 2012)

Used in the early phase of single‐cell cultures in absence of Notch signaling from adjacent supportive cells (Sato et al, 2009; Grabinger et al, 2014)

EGFaActivates RAS/RAF/MEK/ERK signaling pathway (Suzuki et al, 2010; Date & Sato, 2015)Stimulates stem cell migration, proliferation, and inhibits apoptosis (Frey et al, 2004; Suzuki et al, 2010)
PGE2Enhances canonical WNT signaling (Buchanan & DuBois, 2006)Prevents anoikis as well as promotes stem cell survival and proliferation, thereby improving culture efficiency. Stimulates spheroid morphology (Cohn et al, 1997; Joseph et al, 2005)
NicotinamideInhibits the activity of sirtuins (Denu, 2005)Improves ISC maintenance when cultured > 1 week (Sato et al, 2011a). Often used for long‐term human intestinal organoid cultures (Sato et al, 2011a), but can be omitted (Fujii et al, 2015)
Gastrin‐17Not decisively concludedMarginally increases culture efficiency (Sato et al, 2011a)
A83‐01 or SB431542aInhibits TGF‐β signaling (Sato et al, 2011a)Inhibits differentiation and allows human intestinal stem cell cultures to be sustained in the long term (Sato et al, 2011a)
SB202190aInhibits P38 MAPK (Sato et al, 2011a)Inhibits secretory differentiation, increases plating efficiency, and decreases degradation of the EGF receptor (Frey et al, 2006; Sato et al, 2011a; Date & Sato, 2015). Allows human intestinal stem cell cultures to be sustained in the long term (Sato et al, 2011a)
Y‐27632 or thiazovivinInhibition of caspase‐3 (Wu et al, 2015)Prevents anoikis after single‐cell dissociation (Watanabe et al, 2007). Used in the early phase of single‐cell cultures
IL‐22JAK/STAT signaling (Lindemans et al, 2015)ISC proliferation and organoid growth. Can potentially further increase ISC expansion and make EGF redundant (Lindemans et al, 2015)
  • a Mandatory growth medium components for long‐term culturing human intestinal stem cells as organoids.