In Vitro Differentiation and Maturation of Human Embryonic Stem Cell into Multipotent Cells

Table 2

Models for differentiation of embryonic stem cells into multipotent mesenchymal stem cells.

Days in EB formation	Differentiation conditions	Selection	Functional assay	Markers studied	Cells line(s)	Reference
None	Coculture of hESC with OP9 cells in serum-containing media	FACS purification with CD73^+	Differentiation in vitro to osteoblasts, chondrocyte, and adipocytes	Cell surface markers: CD29, CD44, CD54,; CD73, CD90, CD105; D21: mineralization; D17: lipid droplets	Human-H1 and H9	[24]
None	Monolayer direct differentiation in presence of bFGF2 and PDGF AB	FACS purification with CD105^+ and CD24^−	Differentiation in vitro to osteoblasts, chondrocyte and adipocytes	Cell surface markers: CD29, CD44, CD49a,; CD105, CD166; D21: mineralization; D14: lipid droplets; D21: proteoglycans	Human-H1 and HUES9	[25]
None	Spontaneous monolayer differentiated cells from hESC colonies	Serum (10%)	Differentiation in vitro to osteoblasts and adipocytes	Cell surface markers: CD13, CD44, CD71,; CD73, CD105, CD166; After 3 weeks: mineralization^a; D21: lipid droplets	Human-H1	[26]
None	Coculture of hESC with OP9 cells in serum containing media	FACS purification with CD73^+	Differentiation in vitro to osteoblasts, chondrocyte, and myoblasts	Cell surface markers: CD29, CD44, CD54 CD73, CD90, CD105,; CD106, CD166, STRO1	Human-H1 and H9	[27]

Culture medium is α-minimal essential medium or Dulbecco’s modified Eagle’s medium. ^aIn this study they did not analyze the main osteoblastic markers by PCR.
bFGF2: basic fibroblast growth factor-2; CD: cluster of differentiation; OP9: mouse stromal cells.