Human embryonic and rat adult stem cells with primitive endoderm-like phenotype can be fated to definitive endoderm, and finally hepatocyte-like cells.
Stem cell-derived hepatocytes may be an alternative cell source to treat liver diseases or to be used for pharmacological purposes. We developed a protocol that mimics mammalian liver development, to differentiate cells with pluripotent characteristics to hepatocyte-like cells. The protocol supports the stepwise differentiation of human embryonic stem cells (ESC) to cells with characteristics of primitive streak (PS)/mesendoderm (ME)/definitive endoderm (DE), hepatoblasts, and finally cells with phenotypic and functional characteristics of hepatocytes. Remarkably, the same protocol can also differentiate rat multipotent adult progenitor cells (rMAPCs) to hepatocyte-like cells, even though rMAPC are isolated clonally from cultured rat bone marrow (BM) and have characteristics of primitive endoderm cells. A fraction of rMAPCs can be fated to cells expressing genes consistent with a PS/ME/DE phenotype, preceding the acquisition of phenotypic and functional characteristics of hepatocytes. Although the hepatocyte-like progeny derived from both cell types is mixed, between 10-20% of cells are developmentally consistent with late fetal hepatocytes that have attained synthetic, storage and detoxifying functions near those of adult hepatocytes. This differentiation protocol will be useful for generating hepatocyte-like cells from rodent and human stem cells, and to gain insight into the early stages of liver development.
Roelandt
Philip
P
Interdepartmental Stem Cell Institute Leuven, Catholic University Leuven, and Hepatology Department, University Hospitals Leuven, Leuven, Belgium. Philip.roelandt@med.kuleuven.be
Pauwelyn
Karen Ann
KA
Sancho-Bru
Pau
P
Subramanian
Kartik
K
Bose
Bipasha
B
Ordovas
Laura
L
Vanuytsel
Kim
K
Geraerts
Martine
M
Firpo
Meri
M
De Vos
Rita
R
Fevery
Johan
J
Nevens
Frederik
F
Hu
Wei-Shou
WS
Verfaillie
Catherine M
CM
D053687:Adult Stem Cells / Q000166:cytology* / Q000378:metabolism; D000818:Animals; D032701:Biomimetics / Q000379:methods*; D002454:Cell Differentiation; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000378:metabolism; D004707:Endoderm / Q000166:cytology*; D018507:Gene Expression Regulation, Developmental; D022781:Hepatocytes / Q000166:cytology*; D006801:Humans; D051379:Mice; D039902:Multipotent Stem Cells / Q000166:cytology / Q000378:metabolism; D010641:Phenotype; D051381:Rats; D015533:Transcriptional Activation