Reversal of diabetes in mice by intrahepatic injection of bone-derived GFP-murine mesenchymal stem cells infected with the recombinant retrovirus-carrying human insulin gene.
BACKGROUND
The objective of this study was to assess the effect of intrahepatic injection of bone-derived green fluorescent protein (GFP)-transgenic murine mesenchymal stem cells (GFP-mMSCs) containing the human insulin(ins) gene in streptozotocin-induced diabetic mice.


METHODS
GFP-mMSCs were isolated from the bone marrow of GFP transgenic mice, expanded, and transfected with a recombinant retrovirus MSCV carrying the human insulin gene. C57BL/6J mice were made diabetic by an intraperitoneal administration of 160 mg/kg streptozotocin (STZ), followed by intrahepatic injection of transfected GFP-mMSCs. The variations in body weight and the blood glucose and serum insulin levels were determined after cell transplantation. GFP-mMSCs survival and human insulin expression in liver tissues were examined by fluorescent microscopy and immunohistochemistry.


RESULTS
The body weight in diabetic mice that received GFP-mMSCs harboring the human insulin gene was increased by 6% within 6 weeks after treatment, and the average blood glucose levels in these animals were 10.40 +/- 2.80 mmol/l (day 7) and 6.50 +/- 0.89 mmol/l (day 42), respectively, while the average values of blood glucose in diabetic animals without treatment were 26.80 +/- 2.49 mmol/l (day 7) and 25.40 +/- 4.10 mmol/l (day 42), showing a significant difference (p < 0.05). Moreover, secretion of human insulin of GFP-mMSCs in serum and animal liver was detected by radioimmunoassay (RIA) and immunohistochemistry (IHC).


CONCLUSIONS
Experimental diabetes could be relieved effectively for up to 6 weeks by intrahepatic transplantation of murine mesenchymal stem cells expressing human insulin. This study implies a novel approach of gene therapy for type I diabetes.
Xu
Jian
J
Department of General Surgery, the Affiliated Hospital, Nantong University, Nantong, Jiangsu Province, China, 226001.
Lu
Yuhua
Y
Department of General Surgery, the Affiliated Hospital, Nantong University, Nantong, Jiangsu Province, China, 226001.
Ding
Fei
F
Key Laboratory of Neuroregeneration, Nantong University, Nantong, Jiangsu Province, China, 226001.
Zhan
Xi
X
Greenebaum Cancer Center, School of Medicine, University of Maryland, Baltimore, USA.
Zhu
Mingyan
M
Department of General Surgery, the Affiliated Hospital, Nantong University, Nantong, Jiangsu Province, China, 226001.
Wang
Zhiwei
Z
Department of General Surgery, the Affiliated Hospital, Nantong University, Nantong, Jiangsu Province, China, 226001. dxjqhy@yahoo.com.cn.
D000818:Animals; D001842:Bone and Bones; D003921:Diabetes Mellitus, Experimental / Q000139:chemically induced / Q000628:therapy*; D004195:Disease Models, Animal; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins / Q000235:genetics; D006801:Humans; D007004:Hypoglycemic Agents; D007267:Injections / Q000379:methods; D007328:Insulin / Q000235:genetics*; D008099:Liver / Q000378:metabolism; D045164:Mesenchymal Stem Cell Transplantation; D051379:Mice; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D011995:Recombination, Genetic; D012190:Retroviridae / Q000235:genetics*; D016896:Treatment Outcome
D007004:Hypoglycemic Agents; D007328:Insulin; D049452:Green Fluorescent Proteins