Basic Research
Copyright ©2005 Baishideng Publishing Group Inc. All rights reserved.
World J Gastroenterol. Dec 21, 2005; 11(47): 7461-7465
Published online Dec 21, 2005. doi: 10.3748/wjg.v11.i47.7461
Fibroblast growth factor-4 and hepatocyte growth factor induce differentiation of human umbilical cord blood-derived mesenchymal stem cells into hepatocytes
Xin-Qin Kang, Wei-Jin Zang, Li-Jun Bao, Dong-Ling Li,Tu-Sheng Song, Xiao-Li Xu, Xiao-Jiang Yu
Xin-Qin Kang, Tu-Sheng Song, Department of Genetics and Molecular Biology, Medical School of Xi’an Jiaotong University, Xi’an 710061, Shaanxi Province, China
Wei-Jin Zang, Xiao-Li Xu, Xiao-Jiang Yu, Dong-Ling Li, Department of Pharmacology, Key Laboratory of Environment and Genes Related to Diseases of Ministry of Education, School of Medicine, Xi’an Jiaotong University, Xi’an 710061, Shaanxi Province, China
Li-Jun Bao, Department of Obstetrics and Gynecology, First Hospital of Xi’an Jiaotong University, Xi’an 710061, Shaanxi Province, China
Author contributions: All authors contributed equally to the work.
Supported by National Natural Science Foundation of China, No. 30470633 and Doctoral Foundation of Xi’an Jiaotong University, No.DFXJTU2002-16
Correspondence to: Wei-Jin Zang, Box 77, Department of Pharmacology, Key Laboratory of Environment and Genes Related to Diseases of Ministry of Education, School of Medicine, Xi’an Jiaotong University, Xi’an 710061, Shaanxi Province, China. zwj@mail.xjtu.edu.cn
Telephone: +86-29-82655003 Fax: +86-29-82655003
Received: June 13, 2005
Revised: July 1, 2005
Accepted: July 4, 2005
Published online: December 21, 2005
Abstract

AIM: 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 µg/L (t = 2.884, P<0.05) in MSCs cultured with FGF-4 and HGF, and was higher (54.28±3.11 µg/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 µg/mL, and to 3.63±0.30 μg/mL on d 28 (t = 11.748, P<0.01). Urea (4.72±1.03 μmol/L) was detected on d 20 (t = 4.272, P<0.01), and continued to increase to 10.28±1.06 μmol/L on d 28 (t = 9.276, P<0.01). Cells expressed CK-18 on d 16. Glycogen storage was observed on d 24.

CONCLUSION: 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.

Keywords: Mesenchymal stem cell; Differentiation; Hepatocyte