School of Medical Sciences
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Browsing School of Medical Sciences by Subject "Adult stem cells"
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ItemHeralding the Dawn of Cultured Adult Stem Cell Transplantation( 2002-01-01) Vemuganti, Geeta Kashyap ; Balasubramanian, D.Over the last few years, research on stem cells has received much public attention both for its extraordinary potential and for the associated social, legal and public issues. One of the fundamental questions in developmental biology is: how does a single cell - the fertilized cell give rise to a complex, multicellular organism and what are the factors involved during this remarkable development? With that began the quest to define stem cells, identify and isolate the embryonic, fetal and adult stem cells. Then came the question of whether these stem cells derived from various sources can be used to determine the pharmacokinetics of drugs and toxins; and ultimately be used to replace cells in diseased tissues. These questions were answered by, series of experiments on animals followed by clinical trials using cultured adult stem cells, since the latter evoke the least debate on the ethical and legal fronts. Apart from the clinical use of hematopoietic stem cells, the other adult stem cells that have been tested and put to clinical use include: myoblasts for cardiac and skeletal muscle loss or degeneration; cultured beta cells for diabetes mellitus, and cultured limbal epithelial cells for reconstruction of ocular surface in limbal stem cell deficiency. This review briefly describes the concept of stem cells, the regulatory mechanisms that govern them, with special emphasis on stem cells in epithelial tissues and the technique of culturing limbal stem cells that has been developed at our institute.
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ItemIsolation, characterization and differentiation potential of rat bone marrow stromal cells( 2010-04-01) Polisetti, Naresh ; Chaitanya, V. G. ; Babu, Phanithi Prakash ; Vemuganti, Geeta K.Background: Bone marrow mesenchymal cells have been identified as a source of pluripotent stem cells with varying degrees of plasticity in humans. However, there are a few reports on rat-derived cells, which could be good models for the research purpose. We describe here a simple method of establishing the rat bone marrow stromal cells by the principle of adhesion and document their phenotype along with their differentiation potential to other lineages. Materials and Methods: Rat bone marrow stromal cells were isolated by three methods: direct plastic adherence, ficoll hypaque separation and a combination of both. The stromal cells obtained by these methods were characterized by fluorescent activating cell sorting (FACS) for established hematopoietic and non-hematopoietic markers. The cells obtained by combination method (combination of ficoll density gradient centrifugation and plastic adherence) were cultured and serially passaged. Transcriptional confirmation was done by reverse transcription polymerase chain reaction (RT-PCR) for vimentin and collagen type 1 alpha 1. Attempts were made to differentiate the marrow stromal cells into adipocytes, osteocytes and neuronal like cells. Results: Bone marrow samples from 10 rats yielded 4-5 million bone marrow mononuclear cells /ml per femur. Of the three methods tested, a combination method yielded good growth of spindle cells. The cells obtained by combined method showed high percentage of positivity for vimentin, fibronectin and CD90 and negative for hematopoietic markers. Further, RT-PCR confirmed vimentin and collagen type - 1 alpha 1 expression. Oil red O staining and Alizarin red staining confirmed adipocytic and osteogenic differentiation. On immunocytochemical analysis, the cells expressed nestin, -tubulin III, neurofilament and synaptophysin. Conclusion: Adequate quantities of rat marrow stromal cell cultures can be established by a simple method based on adhesion properties. Their phenotypic characteristics and plasticity support the evidence that they are mesenchymal stem cells with a distinct tendency for neural lineage.