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Differentiation of Human Pluripotent Stem Cells Into Pancreatic Beta-Cell Precursors in a 2D Culture System
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Biologia do Desenvolvimento
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JoVE Journal Biologia do Desenvolvimento
Differentiation of Human Pluripotent Stem Cells Into Pancreatic Beta-Cell Precursors in a 2D Culture System

Differentiation of Human Pluripotent Stem Cells Into Pancreatic Beta-Cell Precursors in a 2D Culture System

DOI:
10.3791/63298-v

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10:12 min

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December 16, 2021

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1College of Health and Life Sciences,Hamad Bin Khalifa University (HBKU), Qatar Foundation, 2Diabetes Research Center, Qatar Biomedical Research Institute (QBRI),Hamad Bin Khalifa University (HBKU), Qatar Foundation (QF)

Capítulos

  • 00:05Introduction
  • 01:22Induction of Definitive Endoderm Differentiation in Human Pluripotent Stem Cells (hPSCs) (Stage 1)
  • 02:10Immunofluorescence Analysis of hPSC-Derived Definitive Endoderm (Stage 1)
  • 04:19Generation of the Primitive Gut Tube from hPSCs (Stage 2)
  • 05:50Assessment of Differentiation Efficiency of Generating Pancreatic Progenitors
  • 08:10Results: Comparison between Optimized and Non-Optimized Protocol for Scalable Generation of Pancreatic Progenitors from hPSCs
  • 09:21Conclusion

Summary

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The present protocol describes an enhanced method to increase the co-expression of PDX1 and NKX6.1 transcription factors in pancreatic progenitors derived from human pluripotent stem cells (hPSCs) in planar monolayers. This is achieved by replenishing the fresh matrix, manipulating cell density, and dissociating the endodermal cells.

Tags

Human Pluripotent Stem CellsPancreatic Beta-cell Precursors2D Culture SystemCell TherapyDiabetesPancreatic DevelopmentDefinitive EndodermDifferentiationImmunocytochemistryCHIR-99021

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