TY - JOUR
T1 - Highly efficient differentiation of functional hepatocytes from human induced pluripotent stem cells
AU - Ma, Xiaocui
AU - Duan, YuYou
AU - Tschudy-Seney, Benjamin
AU - Roll, Garrett
AU - Behbahan, Iman Saramipoor
AU - Ahuja, Tijess P.
AU - Tolstikov, Vladimir
AU - Wang, Charles
AU - McGee, Jeannine
AU - Khoobyari, Shiva
AU - Nolta, Jan
AU - Willenbring, Holger
AU - Zern, Mark A
PY - 2013
Y1 - 2013
N2 - Human induced pluripotent stem cells (hiPSCs)hold great potential for useinregenerative medicine, novel drug development, and disease progression/developmental studies. Here, we report highly efficient differentiation of hiPSCs toward a relatively homogeneous population of functional hepa-tocytes. hiPSC-derived hepatocytes (hiHs) not only showed a high expression of hepatocyte-specific proteins and liver-specific functions, but they also developed a functional biotransformation system including phase I and II metabolizing enzymes and phase III transporters. Nuclear receptors, which are critical for regulating the expression of metabolizing enzymes, were also expressed in hiHs. hiHs also responded to different compounds/inducers ofcytochrome P450 as mature hepatocytes do. To follow up on this observation, we analyzed the drug metabolizing capacity of hiHs in real time using a novel ultraperformance liquid chromatography-tandem mass spectrometry. We found that, like freshly isolated primary human hepatocytes, the seven major metabolic pathways of the drug bufuralol were found in hiHs. In addition, transplanted hiHs engrafted, integrated, and proliferated in livers of an immune-deficient mouse model, and secreted human albumin, indicating that hiHs also function in vivo. In conclusion, we have generated a method for the efficient generation of hepatocytes from induced pluripotent stem cells in vitro and in vivo, and it appears that the cells function similarly to primary human hepatocytes, including developing a complete metabolic function. These results represent asignificant step toward using patient/disease-specific hepatocytes for cell-based therapeutics as well as for pharmacology and toxicology studies.
AB - Human induced pluripotent stem cells (hiPSCs)hold great potential for useinregenerative medicine, novel drug development, and disease progression/developmental studies. Here, we report highly efficient differentiation of hiPSCs toward a relatively homogeneous population of functional hepa-tocytes. hiPSC-derived hepatocytes (hiHs) not only showed a high expression of hepatocyte-specific proteins and liver-specific functions, but they also developed a functional biotransformation system including phase I and II metabolizing enzymes and phase III transporters. Nuclear receptors, which are critical for regulating the expression of metabolizing enzymes, were also expressed in hiHs. hiHs also responded to different compounds/inducers ofcytochrome P450 as mature hepatocytes do. To follow up on this observation, we analyzed the drug metabolizing capacity of hiHs in real time using a novel ultraperformance liquid chromatography-tandem mass spectrometry. We found that, like freshly isolated primary human hepatocytes, the seven major metabolic pathways of the drug bufuralol were found in hiHs. In addition, transplanted hiHs engrafted, integrated, and proliferated in livers of an immune-deficient mouse model, and secreted human albumin, indicating that hiHs also function in vivo. In conclusion, we have generated a method for the efficient generation of hepatocytes from induced pluripotent stem cells in vitro and in vivo, and it appears that the cells function similarly to primary human hepatocytes, including developing a complete metabolic function. These results represent asignificant step toward using patient/disease-specific hepatocytes for cell-based therapeutics as well as for pharmacology and toxicology studies.
KW - Hepatocyte differentiation
KW - Induced pluripotent stem cells
KW - Liver regeneration
KW - Stem cell transplantation
UR - http://www.scopus.com/inward/record.url?scp=84878756211&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=84878756211&partnerID=8YFLogxK
U2 - 10.5966/sctm.2012-0160
DO - 10.5966/sctm.2012-0160
M3 - Article
C2 - 23681950
AN - SCOPUS:84878756211
VL - 2
SP - 409
EP - 419
JO - Stem cells translational medicine
JF - Stem cells translational medicine
SN - 2157-6564
IS - 6
ER -