In vitro and in vivo induction of bone formation based on ex vivo gene therapy using rat adipose-derived adult stem cells expressing BMP-7

被引:63
作者
Yang, M
Ma, QJ
Dang, GT
Ma, KT
Chen, P
Zhou, CY [1 ]
机构
[1] Peking Univ, Hlth Sci Ctr, Sch Basic Med Sci, Dept Biochem & Mol Biol, Beijing 100083, Peoples R China
[2] Peking Univ, Hosp 3, Dept Orthoped, Beijing 100871, Peoples R China
基金
中国国家自然科学基金;
关键词
adipose-derived adult stem cells; BMP-7; differentiation; gene therapy;
D O I
10.1080/14653240510027244
中图分类号
Q813 [细胞工程];
学科分类号
摘要
Background Adipose-derived adult stem (ADAS) cells are multipotent cells capable of differentiating into osteoblasts, adipocytes and chondrocytes. The aim of this study was to determine whether BMP-7-expressing ADAS cells would elicit bone formation in vitro and in vivo. Methods ADAS cells were harvested from Lewis rats and transduced with adenovirus carrying the recombinant human bone morphogenetic protein-7 (Ad-BMP-7) gene. Untransduced cells and cells transduced with adenovirus carrying the enhanced green fluorescence protein (Ad-EGFP) gene served as controls. BMP-7 expression was assessed by RT-PCR, immunofluorescence on day 1, and Western blot on days 4, 8 and 12. Alkaline phosphatase (ALP) activity was assayed on days 2, 4, 6, 8, 10 and 12. Osteocalcin production and bone nodule formation were detected by immunohistochemistry and von Kossa stain on day 12. A total of 1 x 10 6 cells mixed with type I collagen were implanted into the subcutaneous pocket in Lewis rat and subjected to histologic analysis 1, 2 and 4 weeks post-implantation. Results The Ad-BMP-7-transduced ADAS cells expressed BMP-7 at both mRNA and protein levels. ALP activity was detected in Ad-BMP-7-transduced cells from day 2 to day 12, peaking on day 8. Osteocalcin production and matrix mineralization further confirmed that these cells differentiated into osteoblasts and induced bone formation in vitro. Histologic examination revealed that implantation of BMP-7-expressing ADAS cells could induce new bone formation in vivo. Discussion ADAS cells would be a promising source of adult autologous stem cells for BMP gene therapy and tissue engineering.
引用
收藏
页码:273 / 281
页数:9
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