Differentiation of human embryonic stem cells to hepatocyte-like cells on a new developed xeno-free extracellular matrix.
Human embryonic stem cells (hESCs) provide a new source for hepatocyte production in translational medicine and cell replacement therapy. The reported hESC-derived hepatocyte-like cells (HLCs) were commonly generated on Matrigel, a mouse cell line-derived extracellular matrix (ECM). Here, we performed the hepatic lineage differentiation of hESCs following a stepwise application of growth factors on a newly developed serum- and xeno-free, simple and cost-benefit ECM, designated "RoGel," which generated from a modified conditioned medium of human fibroblasts. In comparison with Matrigel, the differentiated HLCs on both ECMs expressed similar levels of hepatocyte-specific genes, secreted α-fetoprotein, and metabolized ammonia, showed glycogen storage activity as well as low-density lipoprotein and indocyanine green uptake. The transplantation of hESC-HLCs into the carbon tetrachloride-injured liver demonstrated incorporation of the cells into the host mouse liver and the expression of albumin. The results suggest that the xeno-free and cost-benefit matrix may be applicable in bioartificial livers and also may facilitating a clinical application of human pluripotent stem cell-derived hepatocytes in the future.
Department of Stem Cells and Developmental Biology at the Cell Science Research Center, Royan Institute for Stem Cell Biology and Technology, ACECR, PO Box 19395-4644, Tehran, Iran.
Zahra
Z
Farzaneh
Mohammad
M
Pakzad
Massoud
M
Vosough
Behshad
B
Pournasr
Hossein
H
Baharvand
D000641:Ammonia / Q000378:metabolism; D000818:Animals; D001692:Biological Transport; D002251:Carbon Tetrachloride; D018929:Cell Culture Techniques; D002454:Cell Differentiation; D003094:Collagen; D004338:Drug Combinations; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000637:transplantation; D005109:Extracellular Matrix; D050991:GATA4 Transcription Factor / Q000096:biosynthesis; D005782:Gels / Q000494:pharmacology; D051956:Goosecoid Protein / Q000096:biosynthesis; D051542:Hepatocyte Nuclear Factor 3-beta / Q000096:biosynthesis; D022781:Hepatocytes / Q000166:cytology*; D006801:Humans; D007208:Indocyanine Green / Q000378:metabolism; D057026:Induced Pluripotent Stem Cells / Q000166:cytology* / Q000637:transplantation; D007797:Laminin; D008077:Lipoproteins, LDL / Q000378:metabolism; D008099:Liver / Q000166:cytology / Q000293:injuries / Q000378:metabolism; D051379:Mice; D011509:Proteoglycans; D055748:SOXB1 Transcription Factors / Q000096:biosynthesis; D055760:SOXF Transcription Factors / Q000096:biosynthesis; D000509:alpha-Fetoproteins / Q000096:biosynthesis / Q000378:metabolism
D004338:Drug Combinations; C496502:FOXA2 protein, human; D050991:GATA4 Transcription Factor; C484170:GATA4 protein, human; C497992:GSC protein, human; D005782:Gels; D051956:Goosecoid Protein; D007797:Laminin; D008077:Lipoproteins, LDL; D011509:Proteoglycans; C490906:SOX1 protein, human; C109188:SOX17 protein, human; D055748:SOXB1 Transcription Factors; D055760:SOXF Transcription Factors; D000509:alpha-Fetoproteins; C059485:matrigel; D051542:Hepatocyte Nuclear Factor 3-beta; D000641:Ammonia; D003094:Collagen; D002251:Carbon Tetrachloride; D007208:Indocyanine Green