RESEARCH ARTICLE


AAV Based Ex Vivo Gene Therapy in Rabbit Adipose-derived Mesenchymal Stem/Progenitor Cells for Osteogenesis



Michael Y. Mi 1, 3, Ying Tang 1, Melessa N. Salay 1, Guangheng Li 1, Johnny Huard 1, Freddie H. Fu 1, Christopher Niyibizi 4, Bing Wang 1, 1, *
Molecular Therapy Laboratory, Stem Cell Research Center, Children’s Hospital of Pittsburgh, Department of Orthopaedic Surgery, University of Pittsburgh School of Medicine, Pittsburgh, PA 15261, USA.
Department of Neurology, University of Pittsburgh School of Medicine, Pittsburgh, PA 15261, USA
Harvard College, Harvard University, Cambridge, MA 02138, USA
Department of Orthopaedics and Rehabilitation Pennsylvania State University College of Medicine, Hershey, PA 17033, USA

Article Information


Identifiers and Pagination:

Year: 2009
Volume: 1
First Page: 69
Last Page: 75
Publisher Id: TOSCJ-1-69
DOI: 10.2174/1876893800901010069

Article History:

Received Date: 6/5/2009
Revision Received Date: 24/5/2009
Acceptance Date: 25/5/2009
Electronic publication date: 19/8/2009
Collection year: 2009

Wang et al.; Licensee Bentham Open

open-access license: This is an open access article distributed under the terms of the Creative Commons Attribution 4.0 International Public License (CC-BY 4.0), a copy of which is available at: https://creativecommons.org/licenses/by/4.0/legalcode. This license permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

* Address correspondence to this author at the Molecular Therapy Laboratory, Stem Cell Research Center, Children’s Hospital of Pittsburgh, Department of Orthopaedic Surgery, University of Pittsburgh School of Medicine, Pittsburgh, PA 15261, USA. bingwang@pitt.edu


Abstract

To explore a new source of adult derived stem cells for musculoskeletal tissue repair and regeneration, we isolated rabbit adipose-derived mesenchymal stem/progenitor cells (rAMSCs) from adult rabbit adipose tissue. , the isolated cells were shown to possess the potential to differentiate into cells of mesenchymal lineages that include osteoblasts. The isolated cells were transduced with AAV2 vectors encoding eGFP, human BMP-2, and human TGF-β1 genes prior to implantation into hind limb muscles of SCID mice to assess osteogenesis . The results showed that rAMSCs were more efficiently transduced with GFP by AAV2 than by AAV6. The AAV2-BMP2- and AAV-TGF-β1- transduced rAMSCs showed efficient expression of BMP-2 and TGF-β1 proteins . When the combination of AAV2 BMP2- and AAV-TGF-β1-transduced rAMSCs were implanted into the thigh muscles of SCID mice, the cells osteogenically differentiated toward new bone formation at the implanted sites 15 days after implantation. This study demonstrates that rabbit adipose tissue contains cells with the potential to give rise to osteoblasts, and that the combination of AAV2-BMP-2- and AAV2-TGF-β1-transduced rAMSCs may provide a novel strategy to enhance new bone formation in less than 3 weeks. In addition, the data suggest that the AAV2 vector is suitable for genetic engineering of rAMSCs.

Keywords: AAV, rAMSCs, BMP-2, TGF-β1, osteogenesis.