Table 1.

Directed Differentiation of Mouse PSCs Into Kidney Cells

Study	Starting Substrate	Culture Method	Differentiation Protocol	Differentiation End Point	Findings
Steenhard et al,³⁷ 2005	ROSA26 mESCs		Injection into E12–13 kidneys (3–5 d)	Integration into tubules	Integration into tubular structures expressing LTL and Na⁺/K⁺-adenosine triphosphatase
Kim and Dressier,⁴¹ 2005	ROSA26 mESCs	EB formation	10% FBS without LIF (2 d) 10% FBS + RA/ activin/BMP7 (5 d)	Intermediate mesoderm	Expression of Pax2, WT1, and Lhx1 in EBs by RT-PCR Expression of Pax2 in EBs by IF Integration into LTL⁺ tubular structures when injected into E12.5 kidneys
Kobayashi et al,⁴² 2005	Wnt4-expressing mESCs	EB formation	15% FCS without LIF (2 d) 15% FCS + HGF/ activin (20 d)	AQP2⁺ cells in EBs	Expression of AQP2 by RT-PCR Expression of AQP2 by IF Formation of tubular- like structures in 3D culture
Bruce et al,⁴³ 2006	Pax2-GFP mESCs	EB formation	Serum-free media + BMP4/LIF (16 d)		Expression of Pax2 in EBs by IF
Yamamoto et al,³⁶ 2006	mESCs	EB formation	Teratoma formation (2–4 wk)		Formation of tubular- like structures expressing Pax2 and DBA
Vigneau et al,⁴⁴ 2007	T-GFP mESCs	EB formation	Serum-free media + activin (4 d)	T (Brachyury)⁺ mesoderm	T⁺-GFP cells express cadherin-11, WT1, Pax2, and Wnt4 by RT-PCR T⁺-GFP cells injected into E11.5 kidneys integrate into the nephrogenic zone T⁺-GFP cells injected into newborn mice kidneys integrate into proximal tubules
Nakane et al,⁵⁰ 2009	mESCs with tetracycline- inducible Pax2	EB formation	Pax2 overexpression		Expression of AQP1 in EBs by IF Expression of Pax2 and integrin α8 by RT-PCR
Morizane et al,⁴⁵ 2009	mESCs mi PSCs	EB formation	10% FCS (3 d) 10% FCS + GDNF or BMP7 or activin (15 d)		GDNF and BMP7 induced expression of Pax2 and WT1 Activin induced expression of KSP
Ren et al,⁴⁶ 2010	mESCs	EB formation	15% FCS without LIF (2 d) 15% FCS + activin/ RA (6 d) Conditioned media from UB (10 d)	Renal lineage cells	Expression of Pax2 by IF for IM differentiation Expression of WT1, E-cadherin, POD-1, and Pax2 by IF for renal lineage differentiation
Mae et al,⁴⁹ 2010	mESCs	Monolayer	2% FBS + JAK-inh/ LY294002/ CG1423/RA (8 d)	Intermediate mesoderm	Expression of Osr1 and Pax2 by IF
Nishikawa et al,⁴⁷ 2011	mESCs	Monolayer	10% FBS + activin (2 d) 10% FBS + activin/ BMP4 (2 d) 10% FBS + activin/ BMP4/LiCl (2 d) 10% FBS + activin/ BMP4/LiCl/RA (2 d) Conditioned media from UB and MM cells	UB-conditioned media induced MM markers. MM-conditioned media induced UB markers	Expression of T by RT- PCR for PS differentiation Expression of Pax2 and Lhx1 by RT-PCR for IM differentiation Expression of GDNF, WT, and cadherin-11 by RT-PCR for MM differentiation Expression of HoxB7, Wnt11, and c-Ret9 for UB differentiation
Morizane et al,⁴⁸ 2013	mESCs	EB formation Monolayer	10% FCS + activin (3 d) 10% FCS + activin/ HGF or activin/IGF (15 d)	Ksp⁺ cells	Ksp⁺ cells by FACS Formation of tubular- like structures in Matrigel with expression of kidney markers by IF and RT-PCR Integration of Ksp⁺ cells into tubular structures of E13.5 kidneys
Taguchiet al,³¹ 2013	mESCs hiPSCs	EB formation	Serum-free media + activin (1 d) Serum-free media + BMP4/CHIR (2.5 d) Serum-free media + activin/BMP4/ CHIR/ RA/Y27632 (1 d) Serum-free media + CHIR/FGF9/ Y27632 (2 d)	Metanephric mesenchyme	Nephron precursors derived from OSR1⁺ posterior IM and T⁺ posterior mesoderm Expression of Osr1, WT1, Pax2, Six2, and Sall1 by IF and RT-PCR Co-culture of induced EBs resulted in formation of glomerular-like and tubular-like structures expressing kidney markers Protocol reproducible in hiPSCs with longer time course of differentiation

FACS, fluorescence-activated cell sorting; FBS, fet al bovine serum; FCS, fet al calf serum; GDNF, glial cell–derived neurotrophic factor; KSP, kidney-specific protein; LIF, leukemia-inhibitory factor; RT-PCR, reverse-transcription polymerase chain reaction.