Open AccessIn vitro and in vivo induction of bone formation based on adeno‐associated virus‐mediated BMP‐7 gene therapy using human adipose‐derived mesenchymal stem cells 1
Author(s) -
KANG Yan,
LIAO Weiming,
YUAN Zhenhua,
SHENG Puyi,
ZHANG Longjuan,
YUAN Xiangwei,
LEI Lei
Publication year - 2007
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.1745-7254.2007.00583.x
Subject(s) - mesenchymal stem cell , osteocalcin , bone morphogenetic protein 2 , stem cell , adipose tissue , microbiology and biotechnology , bone morphogenetic protein 6 , in vivo , genetic enhancement , gene delivery , chemistry , bone morphogenetic protein , bone morphogenetic protein 7 , alkaline phosphatase , in vitro , biology , endocrinology , biochemistry , gene , enzyme
Abstract Aim: To determine whether adeno‐associated virus (AAV)‐2‐mediated, bone morphogenetic protein (BMP)‐7‐expressing human adipose‐derived mesenchymal stem cells (ADMS) cells would induce bone formation in vitro and in vivo. Methods: ADMS cells were harvested from patients undergoing selective suction‐assisted lipectomy and transduced with A AV carrying the human BMP‐7 gene. Non‐transduced cells and cells transduced with A AV serotype 2 carrying the enhanced green fluorescence protein gene served as controls. ADMS cells were qualitatively assessed for the production of BMP‐7 and osteocalcin, and subjected to alkaline phosphatase (ALP) and Chinalizarin staining. A total of 2.5×10 6 cells mixed with type I collagen were implanted into the hind limb of severe combined immune‐deficient (SCID) mice and subjected to a histological analysis 3 weeks post implantation. Results: Transfection of the ADMS cells achieved an efficiency of 99% at d 7. Transduction with AAV2‐BMP‐7 induced the expression of BMP‐7 until d 56, which was markedly increased by d 7. The cells were positively stained for ALP. Osteocalcin production and matrix mineralization further confirmed that these cells differentiated into osteoblasts and induced bone formation in vitro. A histological examination demonstrated that implantation of BMP‐7‐expressing ADMS cells could induce new bone formation in vivo. Conclusion: The present in vitro and in vivo study demonstrated that human ADMS cells would be a promising source of autologous mesenchymal stem cells for BMP gene therapy and tissue engineering.