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In vitro Differentiation of Adipose-Derived Mesenchymal Stem Cell into Insulin-Producing Cells

Journal: International Journal of Stem Cell Research and Transplantation (IJST) (Vol.06, No. 01)

Publication Date:

Authors : ;

Page : 287-293

Keywords : Adipose Tissue; Mesenchymal Stem Cell; Islets β-like Cells; ISCT; L-DMEM; FBS; PBS; HRP; DTZl; ELIZA;

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Abstract

Adipose or fatty tissue is similar to bone marrow ontogenetically. Mesenchymal stem cells can be isolated from different types of adipose tissue depots in greater amount than other sources, making them especially suitable for use in regenerative medicine. Adipose tissue was taken from Sudanese donors; the SVF which contains MSCs was isolated using enzymatic and density centrifugation techniques. The AD-MSCs were differentiated to insulin producing cells using three steps protocol. The therapeutic effect of islet β-like cells was determined in vivo using diabetics albino Wister rats. The adherent cells firstly appeared round and spherical in shape, the characteristic shape of MSC was detected after three weeks incubation. The pheno type of these cells showed positives CD34 and CD13 and negatives CD45 and HLADR markers. AD-MSCs were induced into insulin producing cells by a 3-step (15-days) protocol. The differentiated cells were positive for diathizone stain and displayed positive immuno-reactivity to antihuman insulin antibody. Insulin secretion by islet β-like cells in high glucose concentration medium showed positive result with >3.5 u/mlscale reading. The in vivo result of diabetic rats, showed the response in the test group after 24h, one week later the glucose level decreased from 400mg/dl to 96mg/dl (P=0.021), while in the control group the glucose raised up to 600mg/dl. The AD-MSCs can be differentiated into islet β- like cells in vitro and function as insulin producing cells both in vitro and in vivo, these cells are promising source of stem cells for β- cells regeneration.

Last modified: 2018-03-18 02:48:06