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非人类灵长类动物造血干细胞和祖细胞的制备及基因修饰

JoVE Journal
Genetics

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JoVE Journal Genetics

Preparation and Gene Modification of Nonhuman Primate Hematopoietic Stem and Progenitor Cells

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非人类灵长类动物造血干细胞和祖细胞的制备及基因修饰

DOI:

10.3791/58933-v

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11:16 min

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February 15, 2019

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Stefan Radtke, Anai M. Perez, Rasika Venkataraman, Sowmya Reddy, Kevin G. Haworth, Olivier Humbert, Hans-Peter Kiem^2,3, Christopher W. Peterson²

¹Stem Cell and Gene Therapy Program,Fred Hutchinson Cancer Research Center, ²Department of Medicine,University of Washington, ³Department of Pathology,University of Washington

Chapters

00:04Title
00:42Bone Marrow Hemolysis (Day -1)
02:44CD34-enriched Cell Quality Control (Day -1)
05:10CD34+ Hematopoietic Stem and Progenitor Cell (HSPC) Gene Modification and Overnight Recovery (Day 0)
06:33Cell Harvest and Infusion Preparation (Day 1)
07:42Liquid Culture Quality Control (Days 1-5)
09:19Results: Representative HSPC Identification and Analysis
10:34Conclusion

Summary

Automatic Translation

该方案的目标是从启动的骨髓中分离出非人类灵长类动物 cd34⁺细胞, 用慢病毒载体对这些细胞进行基因修饰, 并准备一种产品, 用于注入自体宿主。协议的总长度约为48小时。

Tags

Nonhuman Primate Hematopoietic Stem Cells Progenitor Cells Gene Therapy Bone Marrow Cell Isolation CD34 Magnetic Cell Sorting Flow Cytometry Cell Culture

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