Open AccessRat adipose‐derived stromal cells expressing BMP4 induce ectopic bone formation in vitro and in vivo 1
Author(s) -
LIN Lin,
FU Xin,
ZHANG Xin,
CHEN Lianxu,
ZHANG Jiying,
YU Changlong,
MA Kangtao,
ZHOU Chunyan
Publication year - 2006
Publication title -
acta pharmacologica sinica
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.514
H-Index - 90
eISSN - 1745-7254
pISSN - 1671-4083
DOI - 10.1111/j.1745-7254.2006.00449.x
Subject(s) - mesenchymal stem cell , adipose tissue , stromal cell , von kossa stain , biology , stem cell , pathology , bone morphogenetic protein 4 , alkaline phosphatase , microbiology and biotechnology , bone morphogenetic protein , cancer research , medicine , endocrinology , gene , biochemistry , enzyme
Aim: Bone morphogenetic protein 4 (BMP4) is one of the main local contributing factors in callus formation in the early phase of fracture healing. Adipose‐derived stromal cells (ADSC) are multipotent cells. The present study was conducted to investigate the osteogenic potential of ADSC when exposed to adenovirus containing BMP4 cDNA (Ad‐BMP4). Methods : ADSC were harvested from Sprague‐Dawley rats. After exposure to Ad‐BMP4, ADSC were assessed by alkaline phosphatase activity (ALP) assay, RT‐PCR and von Kossa staining. BMP4 expression was assessed by RT‐PCR, immunofluorescence and Western blot analysis. ADSC transduced with Ad‐BMP4 were directly injected into the hind limb muscles of athymic mice. ADSC Ad‐EGFP(enhanced green fluorescence protein) served as controls. All animals were examined by X‐ray film and histological analysis. Results : The expression of BMP4 was confirmed at both mRNA and protein levels. The expression of the osteoblastic gene, ALP activity and von Kossa staining confirmed that ADSC transduced with Ad‐BMP4 underwent rapid and marked osteoblast differentiation, whereas ADSC transduced with Ad‐EGFP and cells left alone displayed no osteogenic differentiation. X‐ray and histological examination confirmed new bone formation in athymic mice transplanted with ADSC transduced with Ad‐BMP4. Conclusion : Our data demonstrated successful osteogenic differentiation of ADSC transduced with Ad‐BMP4 in vitro and in vivo. ADSC maybe an ideal source of mesenchyme lineage stem cells for gene therapy and tissue engineering.