P-199

Adult stem cells provide a promising potential to generate insulin producing cells in canines.

M.A. Mahgoub, H.M. Samy, N.M. El-Badawy, A.I. Wasfy, A.M. Edris
Ain Shams University, Faculty of Medicin, Cairo, Egypt

Objective: Explore the possibility of the in vitro differentiation of adult stem cells of monocytic origin into functional insulin producing cells & test their therapeutic potency.

Method: The study was conducted on 14 diabetic canine models,2 of them as control .All the procedure was accordant with the animal experimental guidelines of the university. All the canines were under Exogenous insulin therapy till the day of implantation .Monocytes were isolated from peripheral blood of each diabetic canine separately by density gradient centrifugation. The cells were cultured in dedifferentiation medium with interleukin3 and macrophage colony stimulating factor for 6 days, then cultured for an extra 21 days with hepatocyte growth factor & epidermal growth factor to stimulate their differentiation into insulin secreting cells and then examined under inverse microscopy. They were tested for both insulin & glucagon by immuno-histochemistry and for Insulin & C-peptide by radioimmunoassay after glucose challenge test using different glucose concentrations. The cells were injected inside the deltoid muscle of the12 diabetic canines(autologus transplantation).Control canines were injected with culture media free of cells. Follow up was done by measuring the fasting blood glucose level &oral glucose tolerance test.55 days after implantation the grafts were removed &examined for insulin & glucagon by immunohistochemistry.

Results: Typical islets like clustered cells were observed. Immunohistochemistry showed positive insulin & glucagon staining. Glucose challenge test revealed insulin & C- peptide secretion in a glucose dependant manner. The injected cells down regulated the blood glucose level in the 12 diabetic canines .Oral glucose tolerance test showed normal glucose curve in most of the canines. The removed grafts showed positive staining for insulin & glucagon by immunohistochemistry. The control canines remained diabetic.

Conclusion: Functional islet like cells can be differentiated from stem cells of monocytic origin, which may be a new approach for autologus clinical diabetes stem cells therapy.

P-199

Adult stem cells provide a promising potential to generate insulin producing cells in canines.

M.A. Mahgoub, H.M. Samy, N.M. El-Badawy, A.I. Wasfy, A.M. Edris
Ain Shams University, Faculty of Medicin, Cairo, Egypt

Objective: Explore the possibility of the in vitro differentiation of adult stem cells of monocytic origin into functional insulin producing cells & test their therapeutic potency.

Method: The study was conducted on 14 diabetic canine models,2 of them as control .All the procedure was accordant with the animal experimental guidelines of the university. All the canines were under Exogenous insulin therapy till the day of implantation .Monocytes were isolated from peripheral blood of each diabetic canine separately by density gradient centrifugation. The cells were cultured in dedifferentiation medium with interleukin3 and macrophage colony stimulating factor for 6 days, then cultured for an extra 21 days with hepatocyte growth factor & epidermal growth factor to stimulate their differentiation into insulin secreting cells and then examined under inverse microscopy. They were tested for both insulin & glucagon by immuno-histochemistry and for Insulin & C-peptide by radioimmunoassay after glucose challenge test using different glucose concentrations. The cells were injected inside the deltoid muscle of the12 diabetic canines(autologus transplantation).Control canines were injected with culture media free of cells. Follow up was done by measuring the fasting blood glucose level &oral glucose tolerance test.55 days after implantation the grafts were removed &examined for insulin & glucagon by immunohistochemistry.

Results: Typical islets like clustered cells were observed. Immunohistochemistry showed positive insulin & glucagon staining. Glucose challenge test revealed insulin & C- peptide secretion in a glucose dependant manner. The injected cells down regulated the blood glucose level in the 12 diabetic canines .Oral glucose tolerance test showed normal glucose curve in most of the canines. The removed grafts showed positive staining for insulin & glucagon by immunohistochemistry. The control canines remained diabetic.

Conclusion: Functional islet like cells can be differentiated from stem cells of monocytic origin, which may be a new approach for autologus clinical diabetes stem cells therapy.