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Efficient and Scalable Directed Differentiation of Clinically Compatible Corneal Limbal Epithelial Stem Cells from Human Pluripotent Stem Cells
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JoVE 杂志 发育生物学
Efficient and Scalable Directed Differentiation of Clinically Compatible Corneal Limbal Epithelial Stem Cells from Human Pluripotent Stem Cells

Efficient and Scalable Directed Differentiation of Clinically Compatible Corneal Limbal Epithelial Stem Cells from Human Pluripotent Stem Cells

DOI:
10.3791/58279-v

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10:07 min

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October 24, 2018

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, , , ,
1BioMediTech Institute, Faculty of Medicine and Life Sciences,University of Tampere, 2Department of Ophthalmology, SILK, Faculty of Medicine and Life Sciences,University of Tampere

Chapters

  • 00:04Title
  • 00:41Step I: Corneal Induction in Suspension Culture
  • 03:07Step II: Corneal Differentiation in Adherent Culture
  • 04:33Step III: Cryo-banking Human Pluripotent Stem Cell-derived Limbal Epithelial Stem Cells
  • 06:08Step IV: Thawing the Cryopreserved Human Pluripotent Stem Cell-derived Limbal Epithelial Stem Cells
  • 07:25Results: Differentiation of Human Pluripotent Stem Cells to Clinically Compatible Corneal Limbal Epithelial Stem Cells
  • 09:31Conclusion

Summary

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This protocol introduces a simple two-step method for differentiating corneal limbal epithelial stem cells from human pluripotent stem cells under xeno- and feeder cell-free culture conditions. The cell culture methods presented here enable cost-efficient, large-scale production of clinical quality cells applicable to corneal cell therapy use.

Tags

Corneal Limbal Epithelial Stem CellsHuman Pluripotent Stem CellsFeeder-freeEmbryoid Body FormationBasal Induction MediumSB-505124Human Basic Fibroblast Growth FactorLimbal Stem Cell DeficiencyCorneal Cell Replacement Therapy

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