Rotating microgravity-bioreactor cultivation enhances the hepatic differentiation of mouse embryonic stem cells on biodegradable polymer scaffolds.
Embryonic stem (ES) cells are pluripotent cells that are capable of differentiating all the somatic cell lineages, including those in the liver tissue. We describe the generation of functional hepatic-like cells from mouse ES (mES) cells using a biodegradable polymer scaffold and a rotating bioreactor that allows simulated microgravity. Cells derived from ES cells cultured in the three-dimensional (3D) culture system with exogenous growth factors and hormones can differentiate into hepatic-like cells with morphologic characteristics of typical mature hepatocytes. Reverse-transcription polymerase chain-reaction testing, Western blot testing, immunostaining, and flow cytometric analysis show that these cells express hepatic-specific genes and proteins during differentiation. Differentiated cells on scaffolds further exhibit morphologic traits and biomarkers characteristic of liver cells, including albumin production, cytochrome P450 activity, and low-density lipoprotein uptake. When these stem cell-bearing scaffolds are transplanted into severe combined immunodeficient mice, the 3D constructs remained viable, undergoing further differentiation and maturation of hepatic-like cells in vivo. In conclusion, the growth and differentiation of ES cells in a biodegradable polymer scaffold and a rotating microgravity bioreactor can yield functional and organizational hepatocytes useful for research involving bioartificial liver and engineered liver tissue.
The Artificial Liver Lab., Southwest Hospital, The Third Military Medical University, Chongqing, China. yingjiewng@yahoo.com
Yingjie
Y
Wang
Yunping
Y
Zhang
Shichang
S
Zhang
Guangyong
G
Peng
Tao
T
Liu
Yangxin
Y
Li
Dedong
D
Xiang
Michael J
MJ
Wassler
Harnath S
HS
Shelat
Yongjian
Y
Geng
D000418:Albumins / Q000378:metabolism; D000818:Animals; D019149:Bioreactors; D018929:Cell Culture Techniques / Q000295:instrumentation*; D002454:Cell Differentiation / Q000187:drug effects / Q000235:genetics; D049109:Cell Proliferation / Q000187:drug effects; D048430:Cell Shape / Q000187:drug effects; D003577:Cytochrome P-450 Enzyme System / Q000378:metabolism; D058732:Embryoid Bodies / Q000166:cytology / Q000187:drug effects / Q000637:transplantation / Q000648:ultrastructure; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000187:drug effects / Q000648:ultrastructure; D005434:Flow Cytometry; D005455:Fluorescent Antibody Technique; D005786:Gene Expression Regulation / Q000187:drug effects; D019344:Lactic Acid / Q000494:pharmacology*; D008077:Lipoproteins, LDL / Q000378:metabolism; D008099:Liver / Q000166:cytology* / Q000378:metabolism; D051379:Mice; D016513:Mice, SCID; D009928:Organ Specificity / Q000187:drug effects / Q000235:genetics; D011091:Polyesters; D011108:Polymers / Q000494:pharmacology*; D012399:Rotation; D054457:Tissue Scaffolds / Q000737:chemistry; D014893:Weightlessness
D000418:Albumins; D008077:Lipoproteins, LDL; D011091:Polyesters; D011108:Polymers; D019344:Lactic Acid; C033616:poly(lactide); D003577:Cytochrome P-450 Enzyme System