Search academic texts

Information × Registration Number 0211U006665, 0109U004664 , R & D reports Title In vitro induction of pancreatic beta cell phenotype in cryopreserved human adult mesenchymal stem cells derived from bone marrow, skin and adipose tissue popup.stage_title Head Petrenko A.Yu., Registration Date 04-03-2011 Organization Institute for Problems of Cryobiology and Cryomedicine of the National Academy of Sciences of the Ukraine popup.description2 The object of study - human mesenchymal stem cells (MSC) in the induction of differentiation in vitro into insulin-producing cells.The purpose of study - research induction of differentiation in vitro decryopreserved MSCs from adipose tissue (AT), bone marrow (BM) and the skin into insulin-producing cells. The methods of cultivation and cryopreservation, morphological, immunocytochemical analysis were used. Mesenchymal stem cells were isolated from the human BM, stromal-vascular fraction of AT and of skin. After selective cultivation isolated cells demonstrated MSC immunophenotype: CD29 +, CD44 +, CD73 +, CD105 + and CD34 ?, CD38 ?, CD45?. MSCs from of chosen sources were subjected to induction of differentiation in vitro into insulin-producing cells. Foe this aim three different media of differentiation based on DMEM/F12 medium with 10% FCS, 25 mM glucose and 10 mM nicotinamide with and without addition of extract of pancreas of newborn piglets or extract of rat regenerating pancreas were used. The result of differentiation of MSCs into insulin-producing cells were detected by dithizone staining. The presence of clusters and single cells stained with dithizone, was observed in cultures derived from the BM, AT and skin, which allows us to gauge the success of chosen paths of induction of differentiation in all experimental groups.In immunocytochemical studies of fixed cultures of differentiated MSCs have been used antibodies to the hormones of pancreatic islets and dye DAPI. The most of the clusters and single cell cultures after induction in all three media of differentiation stained with antibodies to insulin and proinsulin and only a few clusters were positive for glucagon and somatostatin. Quantitative determination of intracellular and secreted insulin showed that AT MSCs as a result of induced differentiation in the direction of insulin producing cells have the greatest ability to synthesize and secrete insulin, derived BM MSC are characterized by high intracellular content of insulin, and skin derived MSCs practically do not synthesize insulin in response to inducing stimuli. Biotechnology and Cell Biology. Product Description popup.authors Волкова Наталія Олександрівна Грищук Віктор Петрович Мазур Світлана Петрівна Петренко Олександр Юрійович Петренко Юрій Олександрович Петрова Надія Іванівна Скоробогатова Наталія Григорівна popup.nrat_date 2020-04-02 Close