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Retinal Differentiation of Pluripotent Stem Cells: (English)

Retinal Differentiation of Pluripotent Stem Cells: (English)

          
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About the Book

This dissertation, "Retinal Differentiation of Pluripotent Stem Cells" by Debarchana, Sarkar, was obtained from The University of Hong Kong (Pokfulam, Hong Kong) and is being sold pursuant to Creative Commons: Attribution 3.0 Hong Kong License. The content of this dissertation has not been altered in any way. We have altered the formatting in order to facilitate the ease of printing and reading of the dissertation. All rights not granted by the above license are retained by the author. Abstract: The retina is an internal photosensitive neural tunic which absorbs light and prevents it from reflecting back. The light receptors and neurons of the retina are initial processor of visual information. Various anomalies of the retina such as retinitis pigmentosa, cone-rod dystrophy to retinal degenerative diseases cause severe loss of vision since they affect photoreceptors directly or indirectly. Conventional therapies have never been fully successful in restoring vision in such diseases. However current research in stem cell therapies has shown remarkable potential. In this project, induced pluripotent stem cells from mouse were coxed into photoreceptor fate in presence and absence of Dorsomorphin using specific media in a stepwise differentiation process. Dorsomorphin is an inhibitor of Bone Morphogenic Protein (BMP) whose suppression may influence neural differentiation. Studies were done using conventional inverted microscopy and fluorescent microscopy on mouse induced pluripotent stem cells (miPS cells). Immunolabelling techniques involving Pax6, Crx, RPE65, Rhodopsin and Opsin were used to evaluate the advantage of these as markers for stem cells differentiation. Reverse Transcriptase PCR was done to confirm the gene expression on the differentiated cells. Human iPS derived Mesenchymal stem cells were cultured and the effect of different concentrations of Retinoic Acid such as 0mM, 0.1mM and 0.5mM on cell proliferation was tested in both presence and absence of Dorsomorphin. The results revealed both control and Dorsomorphin treated miPS cells successfully differentiated into photoreceptors-like cells as detected by positive staining of Rhodopsin and Opsin. The cells were however negative for Pax6, and very weak staining for RPE and Crx. The presence or absence of Dorsomorphin did not make any difference on miPS differentiation. The same observation was made on differentiating human iPS-MSC where Dorsomorphin did not reveal much effect. However highest cell count of proliferating cells was observed in the subgroups containing 0.1mM Retinoic Acid on Day7 groups as control had an average of 590 317.23 and treatment 1206 .33 114.99 cells, with statistical significance of PIn conclusion, the study reveals that iPS can be another potential stem cell source for therapies of retinal diseases involving photoreceptors where the question of ethical issue is not a problem unlike embryonic stem cells. Also it reveals the concentration of Retinoic Acid most suited for human iPS-MSC cell proliferation. Dorsomorphin did not seem to have much effect on either type of stem cells in terms of promoting photoreceptor differentiation. DOI: 10.5353/th_b5071270 Subjects: Retina - DifferentiationStem cells


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Product Details
  • ISBN-13: 9781360998565
  • Publisher: Open Dissertation Press
  • Publisher Imprint: Open Dissertation Press
  • Height: 279 mm
  • No of Pages: 80
  • Spine Width: 4 mm
  • Width: 216 mm
  • ISBN-10: 136099856X
  • Publisher Date: 26 Jan 2017
  • Binding: Paperback
  • Language: English
  • Series Title: English
  • Weight: 209 gr


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