The Human Skin-Derived Precursors for Regenerative Medicine: Current State, Challenges, and Perspectives

Table 4

The protocols for mesodermal and endodermal differentiation of hSKPs.

Differentiation	Basic medium		Reagent (concentration)	Indication	Ref.
Adipocyte differentiation	DMEM medium		IBMX (0.45 nM), insulin (2.07 μM), dexamethasone (100 nM), amphotecticin B (0.5 μg/ml), Pen/Step (100 units/100 μg/ml), rabbit serum (15%)	Differentiation for 2-3 weeks with medium changed every 3-4 days	[34]
Osteocyte differentiation	DMEM medium		Dexamethasone (100 nM), β-glycerophosphate (10 mM), L-ascorbic acid 2-phosphate (50 μM), Pen/Step (1%), FBS (10%)	Differentiation for 2-3 weeks with medium changed every 3-4 days	[34]
Chondrocyte differentiation	DMEM/F12 (3 : 1) medium		Dexamethasone (100 nM), ascorbic acid (250 mM), bone morphogenetic protein-2 (50 ng/ml), Pen/Step (1%), FBS (10%)	Differentiation for 2-3 weeks with medium changed every 2-3 days	[38]
SMC differentiation	DMEM medium		PDGF-BB (5 ng/ml), TGF-β1 (2.5 ng/ml), FBS (5%)	Differentiation for 3-4 weeks with medium changed every 3 days	[25]
Insulin-producing cell differentiation	Stage 1	High-glucose DMEM/F12 (1 : 1) medium	EGF (20 ng/ml), bFGF (40 ng/ml), B27 (1%), final glucose (17.5 mM)	Culture until forming spheres	[39]
	Stage 2	Low-glucose DMEM medium	db-cAMP (1 mM), RA (1 μM), B27 (1%), 2% FBS, final glucose (5 mM)	Culture for 2 days
	Stage 3	High-glucose DMEM/F12 (1 : 1) medium	Nicotinamide (10 mM), insulin-like growth factor 1 (10 mM), activin-A (2 nM), B27 (1%), 2% FBS, final glucose (17.5 mM)	Culture for up to 1 week
	Stage 4	As stage 3	As stage 3 (except without insulin-like growth factor 1)	Before transferring, cells are trypsinized and resuspended
Hepatocyte differentiation	DMEM/F12 (3 : 1) medium		Day 0: FGF-4 (10 ng/ml); Days 1-2: FGF-4 (10 ng/ml), HGF (30 ng/ml); Days 3–5: FGF-4 (5 ng/ml), HGF (30 ng/ml), 0.5 × ITS; Days 6–8: HGF (30 ng/ml), 0.25 × ITS, dexamethasone (20 μg/l); Days 9–11: HGF (20 ng/ml), dexamethasone (20 μg/l); From day 12 onwards: HGF (20 ng/ml), dexamethasone (20 μg/l), OSM (10 ng/ml)	Differentiation for 3–6 weeks with medium changed every 3 days	[40]

DMEM: Dulbecco’s modified Eagle medium; IBMX: 3-isobutyl-1-methylxanthine; FBS: fetal bovine serum; SMCs: smooth muscle cells; PDGF: platelet-derived growth factor; TGF: transforming growth factor; EGF: epidermal growth factor; bFGF: basic fibroblast growth factor; db-cAMP: dibutyryl cyclic adenosine monophosphate; RA: all-trans retinoic acid; FGF-4: fibroblast growth factor-4; HGF: hepatocyte growth factor; ITS: insulin-transferin-selenite; OSM: oncostatin M.