Developments in cell culture systems for human pluripotent stem cells

Table 1 Primed human pluripotent stem cell culture conditions

	Thomson et al[1]	Xu et al[53]	Xu et al[20]	Vallier et al[16]	Li et al[26]	Ludwig et al[30,80]	Liu et al[52]	Lu et al[33]	Yao et al[117]	Wang et al[23]	Chen et al[17]	Frank et al[118]
FGF2	-	+	+	+	+	+	+	+	+	+	+	+
TGF-β family	-	-	-	Activin	-	TGF-β	-	TGF-β	-	Activin	TGF-β/Nodal	TGF-β/Activin
Serum	FBS	KOSR	KOSR	XF	-	-	-	-	-	-	-	-
Other factors	-	-	Noggin	Insulin	FLT3	Insulin	-	Insulin, BAFF, WNT3A	-	LR3-IGF1, HRG1β	Insulin	Dorsomorphin, IWP-2
Common culture supplements	-	-	NEAA, BME, L-glutamine	BSA, L-glutamine, transferrin, BME, F12, Monothio-glycerol	BME, NEAA, L-glutamine	BSA, BME, transferrin, cholesterol, pipecolic acid, GABA	BME, N2, B27, L-glutamine	BME, transferrin, cholesterol, albumin, L-glutamine	BME, N2, B27, NEAA, BSA-Fraction-V, L-glutamine	BSA-Fraction-V, NEAA, Vc, transferrin, BME, trace elements A, B, C	Vc, transferrin, selenium, NaHCO3	N2, B27, BSA, L-glutamine, NEAA, BME
Base medium	DMEM/F12	DMEM	DMEM/F12	IMDM	XVIVO-10	DMEM/F12	DMEM/F12	DMEM/F12	DMEM/F12	DMEM/F12	DMEM/F12	DMEM/F12
ECM	-	Matrigel	Matrigel	Matrigel	Matrigel	Matrigel	Fibronectin	Matrigel	Matrigel	Matrigel	Matrigel/ Vitronectin	Matrigel
Feeder	MEF	-	-	-	-	-	-	-	-	-	-	-
Markers	Surface markers[119]: SSEA3, SSEA4, TRA1-60, TRA1-81, CD24, CD57, CD90
	Nuclear markers: OCT4, NANOG, SOX2

Table 2 Naïve pluripotent stem cell and extended pluripotent stem cell culture conditions

	Gafni et al[48]	Chan et al[49]	Valamehr et al[50]	Ware et al[51]	Theunissen et al[120]	Guo et al[47]	Ying et al[42]	Yang et al[15]	Yang et al[15]	Yang et al[14]
	Naive hESC	Naive hESC	Naive hESC	Naive hESC	Naive hESC	Naive hESC	Naive mESC	hEPSC	mEPSC	mEPSC
MEK inhibitor	PD0325901	PD0325901	PD0325901	PD0325901	PD0325901	PD0325901	PD0325901	-	-	PD0325901
GSK3 inhibitor	CHIR 99021	BIO	CHIR99021	CHIR99021	IM-12	CHIR99021	CHIR99021	CHIR99021	CHIR99021	CHIR99021
LIF	+	+	+	+	+	+	+	+	+	-
FGF2	+	+	+	+	-	-	-	-	-	-
TGFβ	+	+	-	Activin	Activin	-	-	-	-	-
ROCK Inhibitor	Y27632	Thiazovivin	Thiazovivin	Y27632	Y27632	Y27632	-	Y27632	-	-
Insulin	+	+	-	-	-	+	-	-	-	-
Other factors	JNKi; p38i; PKCi	Dorsomorphin	TGFi	TGFi; FGFRi; HDACi, STAT3i	B-RAFi; LCK/SRCi	PKCi	-	IWR1, dimethindene maleate, minocycline hydrochloride	Dimethindene maleate, minocycline hydrochloride	p38i, JNKi; SRCi; Tanki, WNTi
Base medium	KO DMEM	DMEM/F12	DMEM/F12	DMEM/F12	DMEM/F12/ Neurobasal	DMEM/F12/ Neurobasal	DMEM/F12	DMEM/F12/Neurobasal	DMEM/F12/Neurobasal	DMEM/F12
Medium Supplement	KOSR, NEAA, AlbuMAX I, L-glutamine, BME	TeSR1 components	KOSR, NEAA, BME, L-glutamine	GlutaMAX, KOSR, NEAA, BME, sodium pyruvate	B27/N2, L-glutamine, NEsAA, BME, BSA	N2/B27, L-glutamine, BME, Vc, selenium, putrescine, proges-terone	N2/B27	N2/B27, GlutaMAX, BME, BSA, KOSR	N2/B27, GlutaMAX, BME, BSA, KOSR	BME, GPS, NEAA, KOSR
Feeder	MEF optional	MEF	-	-	MEF	MEF	-	MEF	MEF	MEF
Coating	Gelatin/vitronectin	Matrigel	Matrigel/vitronectin	Matrigel	-	-	Gelatin	-	-	-
Markers	Surface markers[121]: CD7, CD77, CD75, CD130						Oct4, Nanog, Rex1	OCT4, KLF4, NANOG	Oct4, Nanog, Sox2, Sall4	Oct4, Nanog, Sox2, Klf4, Rex1, Esrrb
	Nuclear markers[119]: OCT4, NANOG, SOX2, KLF4,5,17, TFCP2L1, DPPA3,5

mESC: Mouse embryonic stem cell; hEPSC: Human extended pluripotent stem cell; mEPSC: Mouse extended pluripotent stem cell.