Fibroblast growth factor-4 and hepatocyte growth factor induce differentiation of human umbilical cord blood-derived mesenchymal stem cells into hepatocytes.
OBJECTIVE
To investigate the differentiation of human umbilical cord blood (HUCB)-derived mesenchymal stem cells (MSCs) into hepatocytes by induction of fibroblast growth factor-4 (FGF-4) and hepatocyte growth factor (HGF), and to find a new source of cell types for therapies of hepatic diseases.


METHODS
MSCs were isolated by combining gradient density centrifugation with plastic adherence. When HUCB-derived MSCs reached 70% confluence, they were cultured in Iscove modified Dulbecco medium (IMDM) supplemented with 10 mL/L FBS, 20 ng/mL HGF and 10 ng/mL FGF-4. The medium was changed every 4 d and stored for albumin, alpha-fetoprotein (AFP) and urea assay. Expression of CK-18 was detected by immunocytochemistry. Glycogen storage in hepatocytes was determined by PAS staining.


RESULTS
By combining gradient density centrifugation with plastic adherence, we could isolate MSCs from 25.6% of human umbilical cord blood. When MSCs were cultured with FGF-4 and HGF, approximately 63.6% of cells became small, round and epithelioid on d 28 by morphology. Compared with the control, the level of AFP increased significantly from d 12 to 18.20+/-1.16 microg/L (t = 2.884, P<0.05) in MSCs cultured with FGF-4 and HGF, and was higher (54.28+/-3.11 microg/L) on d 28 (t = 13.493, P<0.01). Albumin increased significantly on d 16 (t = 6.68, P<0.01) to 1.02+/-0.15 microg/mL, and to 3.63+/-0.30 microg/mL on d 28 (t = 11.748, P<0.01). Urea (4.72+/-1.03 micromol/L) was detected on d 20 (t = 4.272, P<0.01), and continued to increase to 10.28+/-1.06 micromol/L on d 28 (t = 9.276, P<0.01). Cells expressed CK-18 on d 16. Glycogen storage was observed on d 24.


CONCLUSIONS
HUCB-derived MSCs can differentiate into hepatocytes by induction of FGF-4 and HGF. HUCB-derived MSCs are a new source of cell types for cell transplantation therapy of hepatic diseases.
Kang
Xin-Qin
XQ
Department of Genetics and Molecular Biology, Medical School of Xi'an Jiaotong University, Xi'an 710061, Shaanxi Province, China.
Zang
Wei-Jin
WJ
Bao
Li-Jun
LJ
Li
Dong-Ling
DL
Song
Tu-Sheng
TS
Xu
Xiao-Li
XL
Yu
Xiao-Jiang
XJ
D002454:Cell Differentiation / Q000187:drug effects; D002478:Cells, Cultured; D005312:Fetal Blood / Q000166:cytology*; D051519:Fibroblast Growth Factor 4 / Q000494:pharmacology*; D017228:Hepatocyte Growth Factor / Q000494:pharmacology*; D022781:Hepatocytes / Q000166:cytology*; D006801:Humans; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000187:drug effects
C496445:FGF4 protein, human; D051519:Fibroblast Growth Factor 4; D017228:Hepatocyte Growth Factor