Perspectives of pluripotent stem cells in livestock

Table 2 Most recent examples of induced pluripotent stem cells successfully generated from livestock

Species	Cell type	Culture medium and condition	Expression of pluripotency markers	Long-term culture (passage)	Karyotype	In vitro differentiation to EBs	In vivo differentiation to teratomas	Germline transmission	Ref.
Cattle	Fetal fibroblasts	DMEM/F-12, 20% KSR, 1 mM l-glutamine, 0.1 mM NEAAs, 0.1 mM mercaptoethanol, 100 U/mL penicillin, 100 μg/mL streptomycin, 8 ng/mL bFGF, 1000 U/mL hLIF on MEF at 37℃ and 5% CO2	Yes	40	Normal	Yes	Yes	No	[58]
	Fetal fibroblasts	KO-DMEM, 15% FBS, 2 mM L-glutamine, 1% NEAAs, 0.1 mM β-mercaptoethanol, 106 U/mL hLIF, 10 ng/mL bFGF on STO at 37℃ and 5% CO2	Yes	50	Normal	Yes	Yes	No	[130]
Buffalo	Fetal fibroblasts	DMEM high glucose, 20% ESC-FBS, 2 mM l-glutamine, 1% NEAAs, 0.1 mM β-mercaptoethanol, 10 ng/mL bFGF, 10 ng/mL LIF on MEF at 37℃ and 5% CO2	Yes	10	Normal	Yes	Yes	No	[65]
	Fetal fibroblasts	DMEM/F-12, 20% KSR, 0.1 mM NEAAs, 1 mM L-glutamine, 0.1 mM mercaptoethanol, 100 U/mL penicillin, 100 μg/mL streptomycin, 10 ng/mL bFGF, 1000 U/mL hLIF on gelatine at 37℃ and 5% CO2	Yes	15	Normal	Yes	No	No	[66]
Ovine	Embryonic fibroblasts	DMEM, 20% FBS, 1% ITS, 0.1 mM 2-β mercaptoethanol, 1 mM NEAAs, 2 mM glutamine, 4 ng/mL bFGF, 1000 U/mL mLIF on MEFs at 37℃ and 5% CO2	Yes	17	Normal	Yes	Yes	Yes (formation of ICM in tetraploid)	[50]
	Embryonic fibroblasts	KO-DMEM, KSR, 0.1 mM NEAAs, 2 mM L-glutamine, 0.1 mM 2-mercaptoethanol, 8 ng/mL hFGF2, 1000 U/mL mLIF on SNL at 37℃ and 5% CO2	Yes	23	Normal	Yes	Yes	Yes (live-born chimeric lambs)	[67]
Caprine	Fetal fibroblasts	DMEM/F12, 20% KSR, 1 mM L-glutamine, 0.1 mM 2-mercaptoethanol, 1% NEAAs, 2% sodium bicarbonate solution, 1000 IU/mL 2i/LIF, 4 ng/mL bFGF on STO at 37℃ and 5% CO2	Yes	30	Normal	Yes	Yes	No	[282]
	Embryonic fibroblasts	KO-DMEM, 20% KSR, 1% NEAA, 1% L-glutamine, 0.1 mM EAA, 1% penicillin /streptomycin, 10 ng/mL FGF2 on GEF at 37℃ and 5% CO2	Yes	22	Normal	Yes	No	No	[283]
Porcine	Embryonic fibroblasts and microvascular pericyte cells	LCDMV medium contains 50% neurobasal medium, 50% DMEM/F12, 1 × N2, 0.5 × B27, 5% KSR, 10 ng/mL LIF, 1 μM CHIR99021, 2 μM (S)-(+)-dimethindene maleate, 2 μM minocycline hydrochloride, 40 μg/mL vitamin C on MEF at 37℃ and 5% CO2	Yes	28	Normal	Yes	Yes	Yes (chimeric formation in post-implantation pig conceptuses)	[284]
	Sertoli cells	DMEM/F12, 10% KSR, 10% FBS, 1 mM l-glutamine, 1 mM antibiotic, 1% NEAAs, 0.1 mM β-mercaptoethanol, 10 ng/mL bFGF, 10 ng/mL hLIF on MEF at 37℃ and 5% CO2	Yes	50	Normal	Yes	Yes	No	[285]
Equine	Fetal fibroblasts	DMEM/F12, 20% KSR, 10 ng/mL bFGF, 1% penicillin/streptomycin, 10 ng/mL hLIF on MEF at 37℃ and 5% CO2	Yes	25	--	Yes	No	No	[286]
	Fetal fibroblasts	DMEM, 20% FBS or KO-DMEM, 20% KSR, 2 mM l-glutamine, 0.1 mM β-mercaptoethanol, 0.1 mM MEM- NEAAs, 1% penicillin–streptomycin, 8 ng/mL hbFGF, 1000 U/mL hLIF on SNL at 37℃ and 5% CO2	Yes	30	Normal	Yes	Yes	No	[61]
Canine	Embryonic fibroblasts	Serum-free N2B27-based medium, 4 ng/mL hbFGF on MEF at 37℃ and 5% CO2	Yes	50	Normal	Yes	No	No	[287]
	Fetal fibroblasts	KO-DMEM/F12, 20% KSR, 2 mM L-glutamine, 0.1 mM NEAAs, 0.1 mM β-mercaptoethanol, 0.1 mM bFGF on MEF at 37℃ and 5% CO2	Yes	15	Normal	Yes	Yes	No	[288]

DMEM: Dulbecco's modified Eagle's medium; KO-DMEM: Knockout Dulbecco's modified Eagle's medium; DMEM/F12: Dulbecco's modified Eagle's medium/nutrient mixture F-12;‎ NEAAs: Non-essential amino acids; bFGF: Basic fibroblast growth factor; hLIF: Human leukemia inhibitory factor; MEF: Mouse embryonic fibroblast; KSR: Knock-out serum replacement; FBS: Fetal bovine serum; LIF: Leukemia inhibitory factor; ESC-FBS: Embryonic stem cells-fetal bovine serum; ITS: Insulin–transferrin–selenium; mLIF: Mouse leukemia inhibitory factor; hFGF2: Human fibroblast growth factor 2; EAAs: Essential amino acids; hbFGF: Human basic fibroblast growth factor; STO: Sandos inbred mouse-derived 6-thioguanine-and ouabain-resistant.