Bone Marrow-Derived Stem Cells Can Differentiate into Retinal Cells in Injured Rat Retina<br />By Anna Begelfer-Ostrovski<...
Bone marrow-<br />spongy, fatty tissue houses stem cells.<br />located inside a few large bones.<br />It’s stem cells tran...
Bone marrow cells (BMCs) differentiate into various cells:<br />hepatocytes<br />endothelial cells of the blood vessel <br...
This experiment:<br />examined whether bone marrow stem cells can differentiate into retinal neural cells in adult rats.<b...
Experiment overview:<br />Collection of BMC’s stem cells.<br />Injury of Rat’s eye.<br />Injection of BMC’s stem cells<br ...
Materials and Methods<br />Preparation of Stem Cell-Enriched BMCs for grafting:<br />5-fluorouracil injected into the peri...
Materials and Methods<br />Rats and Grafting Procedure<br />rats were anesthetized with pentobarbital sodium.<br />pupils ...
Materials and Methods<br />Injury:<br />eyeballs were perforated at the equator with a 30-gauge needle<br />retina was inj...
Materials and Methods<br />Post injury: <br />20 μl of the cell suspension, containing 200,000 cells, were slowly injected...
Materials and Methods<br />Tissue Sectioning<br />The animals were sacrificed 2 weeks after injection<br />eyes were enucl...
Results: <br />BMCs had been built into the host retina in 83% of the experimental eyes (5/6). <br />Saline treated retina...
Image I:<br />Saline injection<br />Proliferated due to BMC’s injection<br />Distribution of BMC’s using green fluorescenc...
Image I:<br />The grafted cells( green fluorescence) were distributed around the injured sites.<br />Grafted cells were ob...
Image II<br />Immunohistochemistry on Sections after Transplantation<br />
Image II <br />Injected cells labeled with expressed green fluorescence. <br />Astrocytes, carbindin, vimentin, and rhodop...
 Discussion:<br />stem cell-enriched BMCs injected into injured eyes differentiated into retinal neural cells in vivo.<br ...
Conclusion:<br /> stem cell-enriched BMCs have the ability to differentiate into retinal neural cells.<br />Injection of s...
Take home message: <br />injection of BMC’s into the eye can potentially rescue injured retinal tissue, even in humans.<br />
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Bone Marrow Derived Stem Cells Can Differentiate Into Retinal

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Bone Marrow Derived Stem Cells Can Differentiate Into Retinal

  1. 1. Bone Marrow-Derived Stem Cells Can Differentiate into Retinal Cells in Injured Rat Retina<br />By Anna Begelfer-Ostrovski<br />Minoru Tomita1,2, <br />Yasushi Adachi1,2,3,<br />Haruhiko Yamada2, <br />Kanji Takahashi2,<br />Katsuji Kiuchi2,5, <br />Haruki Oyaizu1, <br />Kazuya Ikebukuro1, <br />Hiroyuki Kaneda1, <br />Miyo Matsumura2,3,4, <br />Susumu Ikehara M.D.1,3,<br /> <br />
  2. 2. Bone marrow-<br />spongy, fatty tissue houses stem cells.<br />located inside a few large bones.<br />It’s stem cells transform themselves into white and red blood cells and platelets.<br />essential for immunity and circulation<br />
  3. 3. Bone marrow cells (BMCs) differentiate into various cells:<br />hepatocytes<br />endothelial cells of the blood vessel <br />epithelial cells of the stomach, esophagus, small intestine, large intestine and bronchus<br />cardiac muscle <br />skeletal muscle<br />neural cells <br />
  4. 4. This experiment:<br />examined whether bone marrow stem cells can differentiate into retinal neural cells in adult rats.<br />
  5. 5. Experiment overview:<br />Collection of BMC’s stem cells.<br />Injury of Rat’s eye.<br />Injection of BMC’s stem cells<br />Sectioning of the eye <br />Data Analysis<br />Conclusion<br />
  6. 6. Materials and Methods<br />Preparation of Stem Cell-Enriched BMCs for grafting:<br />5-fluorouracil injected into the peritoneal cavity 4-week old male rat (inside abdominal cavity)<br />BMCs were obtained from femurs and tibias (leg)<br /> Fractionated by density centrifugation using Lymphoprep™ density solution <br />collected and labeled. <br />
  7. 7. Materials and Methods<br />Rats and Grafting Procedure<br />rats were anesthetized with pentobarbital sodium.<br />pupils were dilated with 0.5% tropic amide <br />the corneas were further anesthetized with drops of 0.4% oxybuprocainehydrochlorid<br />
  8. 8. Materials and Methods<br />Injury:<br />eyeballs were perforated at the equator with a 30-gauge needle<br />retina was injured by scratching  (under a surgical microscope)<br />special care was taken to injure the whole layer of the retina <br />
  9. 9. Materials and Methods<br />Post injury: <br />20 μl of the cell suspension, containing 200,000 cells, were slowly injected into the intravitreous space.<br />6 rats, 6 injected eyes with BMC’s stem cells.<br />Control-20 μl of normal saline solution injected into the same place of injured eyes. <br />
  10. 10. Materials and Methods<br />Tissue Sectioning<br />The animals were sacrificed 2 weeks after injection<br />eyes were enucleate and prepared for cutting. <br />Samples were stained and observed under confocal microscope. <br />
  11. 11. Results: <br />BMCs had been built into the host retina in 83% of the experimental eyes (5/6). <br />Saline treated retina showed progressing damage. <br />BMC’s treated cells showed cell proliferation.<br />
  12. 12. Image I:<br />Saline injection<br />Proliferated due to BMC’s injection<br />Distribution of BMC’s using green fluorescence<br />C without Flourescence<br />
  13. 13. Image I:<br />The grafted cells( green fluorescence) were distributed around the injured sites.<br />Grafted cells were observed at the site of injury and at normal retinal structure.<br />Most of the grafted cells were in the injured site<br />BMCs had migrated into the retina, followed by proliferation and differentiation into retinal neural cells.<br />
  14. 14. Image II<br />Immunohistochemistry on Sections after Transplantation<br />
  15. 15. Image II <br />Injected cells labeled with expressed green fluorescence. <br />Astrocytes, carbindin, vimentin, and rhodopsin, red fluorescence. <br />Incorporated bone marrow-derived cells, yellow fluorescence.<br /> Incorporated bone marrow cells had differentiated into retinal cells. <br />
  16. 16.  Discussion:<br />stem cell-enriched BMCs injected into injured eyes differentiated into retinal neural cells in vivo.<br />Injection of retinal tissue of newborn and brain-derived cell lines are other methods that did not bring same results. <br />
  17. 17. Conclusion:<br /> stem cell-enriched BMCs have the ability to differentiate into retinal neural cells.<br />Injection of stem cell-enriched BMCs into the retina would help repair damaged retinal cells.<br />
  18. 18. Take home message: <br />injection of BMC’s into the eye can potentially rescue injured retinal tissue, even in humans.<br />

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