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Annals of Clinical & Laboratory Science 36:127-136 (2006)
© 2006 Association of Clinical Scientists

Human Bone Marrow Mesenchymal Stem Cells Transfected with Human Insulin Genes Can Secrete Insulin Stably

Yuhua Lu, Zhiwei Wang and Mingyan Zhu
Department of General Surgery, Affiliated Hospital of Nantong University, Nantong, Jiangsu, China

Address correspondence to Dr. Zhiwei Wang, Depatment of General Surgery, Affiliated Hospital of Nantong University, Nantong, Jiangsu Province, China, 2226001; tel 86 138 6281 5432; fax 86 513 8610 2729; e-mail lyh_sigma{at}sohu.com.

Beta-cell replacement therapy by pancreatic islet transplantation has become a promising treatment for type 1 diabetes. However, the limited supply of human islet tissue prevents this therapy from being widely used to treat patients with type 1 diabetes. In order to obtain insulin-secreting cells, retrovirus vector pLNCX was used to transfer the human insulin gene into human bone marrow mesenchymal stem cells (hMSCs). The hMSCs were isolated from bone marrow of healthy volunteers and were expanded in vitro. The reverse transcriptase-polymerase chain reaction (RT-PCR) was used to amplify the insulin DNA fragment from a healthy pancreas sample. The recombinant vector pLNCX-Ins was constructed by cloning the insulin DNA fragment into retrovirus vector pLNCX. After being packaged by BD RetroPack PT67 packaging cells, the virus that contained the insulin gene was used to infect hMSCs. Transcription and expression of the insulin gene in transfected hMSCs were examined by RT-PCR and immunofluorescence. The transfected hMSCs stably secreted insulin into culture media for >3 weeks. Thus, insulin gene-transfected hMSCs can secrete insulin and provide a new way to cope with the shortage of beta cells for therapy of type 1 diabetes.

Keywords: mesenchymal stem cells, bone marrow, retroviral vector, gene therapy, insulin, type 1 diabetes






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