Hepatic differentiation of human embryonic stem cells on microcarriers.
Translation of stem cell research to industrial and clinical settings mostly requires large quantities of cells, especially those involving large organs such as the liver. A scalable reactor system is desirable to ensure a reliable supply of sufficient quantities of differentiated cells. To increase the culture efficiency in bioreactor system, high surface to volume ratio needs to be achieved. We employed a microcarrier culture system for the expansion of undifferentiated human embryonic stem cells (hESCs) as well as for directed differentiation of these cells to hepatocyte-like cells. Cells in single cell suspension were attached to the bead surface in even distribution and were expanded to 1×10(6)cells/ml within 2 days of hESC culture with maintenance of the level of pluripotency markers. Directed differentiation into hepatocyte-like cells on microcarriers, both in static culture and stirred bioreactors, induced similar levels of hepatocyte-like cell differentiation as observed with cells cultured in conventional tissue culture plates. The cells expressed both immature and mature hepatocyte-lineage genes and proteins such as asialoglycoprotein receptor-1 (ASGPR-1) and albumin. Differentiated cells exhibited functional characteristics such as secretion of albumin and urea, and CYP3A4 activity could be detected. Microcarriers thus offer the potential for large-scale expansion and differentiation of hESCs induced hepatocyte-like cells in a more controllable bioreactor environment.
Stem Cell Institute Leuven, Department of Development and Regeneration, Cluster Stem Cell Biology and Embryology, KU Leuven Medical School, Belgium.
Yonsil
Y
Park
Stem Cell Institute Leuven, Department of Development and Regeneration, Cluster Stem Cell Biology and Embryology, KU Leuven Medical School, Belgium.
Yemiao
Y
Chen
Stem Cell Institute Leuven, Department of Development and Regeneration, Cluster Stem Cell Biology and Embryology, KU Leuven Medical School, Belgium.
Laura
L
Ordovas
Stem Cell Institute Leuven, Department of Development and Regeneration, Cluster Stem Cell Biology and Embryology, KU Leuven Medical School, Belgium. Electronic address: catherine.verfaillie@med.kuleuven.be.
Catherine M
CM
Verfaillie
D000418:Albumins / Q000235:genetics / Q000378:metabolism; D037263:Asialoglycoprotein Receptor / Q000235:genetics / Q000378:metabolism; D015415:Biomarkers / Q000378:metabolism* / Q000652:urine*; D019149:Bioreactors; D018929:Cell Culture Techniques / Q000379:methods*; D002454:Cell Differentiation / Q000502:physiology; D048708:Cell Growth Processes; D002460:Cell Line; D051544:Cytochrome P-450 CYP3A / Q000235:genetics / Q000378:metabolism; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000378:metabolism; D022781:Hepatocytes / Q000166:cytology* / Q000378:metabolism; D006801:Humans
C546927:ASGR1 protein, human; D000418:Albumins; D037263:Asialoglycoprotein Receptor; D015415:Biomarkers; D051544:Cytochrome P-450 CYP3A; C510163:CYP3A4 protein, human
Bioreactor; Hepatic differentiation; Human embryonic stem cells; Microcarrier; Suspension culture system