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Leukodepletion Filters-Derived CD34+ Cells As a Cell Source to Study Megakaryocyte Differentiation and Platelet Formation
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Journal JoVE Biologie
Leukodepletion Filters-Derived CD34+ Cells As a Cell Source to Study Megakaryocyte Differentiation and Platelet Formation

Leukodepletion Filters-Derived CD34+ Cells As a Cell Source to Study Megakaryocyte Differentiation and Platelet Formation

DOI:
10.3791/62499-v

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06:57 min

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May 20, 2021

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, , , , ,
1BPPS UMR-S 1255, FMTS,Université de Strasbourg, INSERM, EFS Grand Est

Chapitres

  • 00:04Introduction
  • 01:01Leukoreduction Filter (LRF) Back-Flushing Modality Preparation
  • 01:48PBMC Collection
  • 02:47CD34+ Cell Selection
  • 04:01Evaluation of CD34+ Cell Purity
  • 04:35CD34+ Cell Freezing
  • 05:07Results: Mature Megakaryocyte Differentiation and Platelet Formation
  • 06:16Conclusion

Summary

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Traduction automatique

This protocol describes in detail all the steps involved in obtaining leukofilter-derived CD34+ hematopoietic progenitors and their in vitro differentiation and maturation into proplatelet-bearing megakaryocytes that are able to release platelets in the culture medium. This procedure is useful for in-depth analysis of cellular and molecular mechanisms controlling megakaryopoiesis.

Tags

Leukodepletion FiltersCD34+ CellsMegakaryocyte DifferentiationPlatelet FormationHematopoietic ProgenitorsMegakaryopoiesisPBMC CollectionDensity Gradient CentrifugationCD34+ Cell SelectionFC Receptor BlockingCD34 Microbeads

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