International Journal of Pediatrics

Isolation, Characterization and Differentiation of Rat Adipose Tissue Derived Mesenchymal Stem Cells

Seyed Javad Hoseini; Hamed Ghazavi; Ahmad Ghorbani; Hamid Reza Sadeghian; Majid Ghayour Mobarhan; Elaheh Mahdipour

Volume 2, Issue 2.3 , May 2014, , Pages 41-41

https://doi.org/10.22038/ijp.2014.2694

Abstract

Introduction: Mesenchymal stem cells have the potential of self-renewal and differentiation into different cell types, including blood cells, heart, nerves and cartilage, ... Read More Introduction: Mesenchymal stem cells have the potential of self-renewal and differentiation into different cell types, including blood cells, heart, nerves and cartilage, and have unlimited power for division. These cells can be obtained from cord, before implantation from fertilized cells and also from various tissues of adults although the differentiation power and the ability to reproduce other cells are different. The use of these cells in the biological sciences, embryology and genetics are considered further.The aims of this study are extraction and characterization of stem cells with respect to differentiation of these cells into bone and fat cells in order to investigate the potential use of them in animal experimental models of stroke and heart attack and then in the case of the proper response at the bedside. Methods: An incision was made in the abdomen and about one gram of fat tissue was removed. After splitting under the hood, pieces of fat transferred into a sterile falcon tube and collagenase was added. The tube was put inside a water bath (37 ° C) for digestion fat components. After centrifugation the cells were extracted and cultured. After the third passage, the cells were placed in the appropriate ostogenic and adipogenic differentiation culture. To ensure the accuracy of the extraction method, the cell surface markers of the stem cells derived from human adipose tissue were analyzed by flowcytometry. Results: Staining with Oil Red and Alizarin Red showed that stem cells extracted by the aforementioned method have the ability to differentiate into bone and fat cells respectively. The results of flocytometry showed that CD34 and CD44 markers were negative and CD45 and CD105 were positive consistent with the immunological properties of Mesenchyma stem cells. Conclusions: This study showed that the mesenchymal stem cells derived from adipose tissue have the potential to differentiate into other cell types and due to the aims specified for continuation of this research, these cells can be used in the animal experimental models of stroke and myocardial infarction. Keywords: Adipose tissue, Mesenchymal stem cells,Rat.

Isolation of Adipose Tissue Stem Cells with Organ Culture Method

Ahmad Ghorbani; Seyed Amir Jalali; Masoumeh Varedi

Volume 2, Issue 2.3 , May 2014, , Pages 53-53

https://doi.org/10.22038/ijp.2014.2716

Abstract

Background Isolation of adipose tissue mesenchymal stem cells (MSCs) with current enzymatic methods has some limitations. For example, it is costly and time-consuming and results ... Read More

Rat Bone Marrow Mesenchymal Stem Cell Differentiation to Insulin Producing Cells and Evaluation their Responses in Vitro and in Vivo

Seyyed Abbas Zojaj; Reza Shafiee-Nick; Ahmad Ghorbani

Volume 2, Issue 2.3 , May 2014, , Pages 64-64

https://doi.org/10.22038/ijp.2014.2727

Abstract

Background In recent years, many researchers haveattempted to cure diabetes by using stem cells technology. Stem cells from different sources have capabilityto differentiateinto ... Read More Background In recent years, many researchers haveattempted to cure diabetes by using stem cells technology. Stem cells from different sources have capabilityto differentiateinto insulin producing cells (IPCs) by different methods. The obstaclesof these methods aretheirexpensive materials and complexity ofmethodswhichare practicallydisadvantagesfor producing enough transplantableIPCs that can cure a diabetic patient. The aim of present study was to test a method for isolation and differentiation of rat bone marrow mesenchymal stem cells (BMSC)into IPCs which is simple method and produce abundant IPCs. Methods: Adult male Wistar rats 4-6 weeks age and 200-250 g weight were used. Bone marrow was isolated from femoral bone under asepticconditionand cultured as monolayerin DMEM in a density of 1×106/well at 37ºC and 5% CO2 for72 h. After three passages, the cells were differentiated into adiposities and osteocyte cells which approved using oil red and alizarin red staining, respectively. To produce IPCs, BMSC was cultured in low glucose DMEM, with 1% DMSO and without FBS for three days on collagenated coverslips. After three days, theculture medium was changedto DMEM containing 25 mM glucose plus10%FBS and incubated for 7 days. For assessment of insulin secretion capabilityof IPCs, the cells were incubated in DMEM containing5.5 or 25 mM glucose with or without IBMX for two hours. Samples from the medium were taken for subsequent insulin assay For in vivo assay, the mice were made diabetic by injection 200 mg/kgstreptozocinintraperitoneally,which produced a fasting blood sugar about 450 mg/dl. For each mouse, 105 IPCs were injectedintraperitonally. Control mice were injected with 100 µl normal saline.The mice blood sugars were checked every 3 days with a glocoumeter device. Results: The content of secreted insulin in 5.5 m M glucose concentration was about 40miu/ml that raised to72 miu/ml with incubation with 25 mM glucose. The insulin secretion was potentiated by IBMX which increased to 104miu/ml. One week after IPC transplantation to diabetic mice, fasting blood sugar was started to decrease. After two weeks of transplantation, the blood glucose was fallen to 265 mg/dL comparing to control mice which their fasting blood sugar was about 450 mg/dl until end of study. Conclusion: In this study we introduced a simple, valuable and low cost method to produce insulin producing cells from bone marrow mesenchymal stem cells which could be translated into human clinical trials. Keywords: Mesenchymal stem cells, Transplantation, Diabetes.

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Author = Ahmad Ghorbani

Isolation, Characterization and Differentiation of Rat Adipose Tissue Derived Mesenchymal Stem Cells

Abstract

Isolation of Adipose Tissue Stem Cells with Organ Culture Method

Abstract

Rat Bone Marrow Mesenchymal Stem Cell Differentiation to Insulin Producing Cells and Evaluation their Responses in Vitro and in Vivo

Abstract