Identification of Nonhuman Primate Hematopoietic Stem and Progenitor Cells.
Identification of Nonhuman Primate Hematopoietic Stem and Progenitor Cells.
复制标题
非人灵长类造血干细胞和祖细胞的鉴定。
DOI：
10.1007/978-1-0716-2679-5_6
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发表时间：
2023
期刊：
影响因子：
--
通讯作者：
Kiem,Hans-Peter
中科院分区：
文献类型：
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作者：
Radtke,Stefan;Kiem,Hans-Peter
造血干细胞（HSC）基因治疗/编辑和移植方案的临床前开发经常在大型动物模型如非人灵长类动物（NHP）中进行。大多数试剂和药物的生理学、大小和预期寿命以及交叉反应性的相似性允许开发快速转化为临床应用的治疗策略。特别是在20世纪90年代末观察到HSC基因治疗的不良事件之后，进行自体移植和长期随访动物的能力使NHP成为一种非常有吸引力的模型，用于测试HSC基因治疗的效率、可行性、新HSC介导的基因转移的安全性/该方案描述了一种在标本或干细胞内表型表征功能不同的NHPHSPC亚群的方法。来自三种不同NHP物种的产品。程序基于对人和NHP之间具有交叉反应性的细胞表面标志物的流式细胞术评估，以允许立即进行临床转化。该方案已成功用于猪尾猕猴、食蟹猴和恒河猴移植用富集、培养和基因修饰的NHPCD 34+造血干细胞和祖细胞（HSPC）以及分选纯化的CD 34亚群的质量控制。它还允许对移植后长期随访期间采集的原始标本进行纵向评估，以监测骨髓干细胞室的归巢、植入和重建。
The preclinical development of hematopoietic stem cell (HSC) gene therapy/editing and transplantation protocols is frequently performed in large animal models such as nonhuman primates (NHPs). Similarity in physiology, size, and life expectation as well as cross-reactivity of most reagents and medications allows for the development of treatment strategies with rapid translation to clinical applications. Especially after the adverse events of HSC gene therapy observed in the late 1990s, the ability to perform autologous transplants and follow the animals long-term make the NHP a very attractive model to test the efficiency, feasibility, and safety of new HSC-mediated gene-transfer/editing and transplantation approaches.This protocol describes a method to phenotypically characterize functionally distinct NHPHSPC subsets within specimens or stem cell products from three different NHP species. Procedures are based on the flow-cytometric assessment of cell surface markers that are cross-reactive in between human and NHP to allow for immediate clinical translation. This protocol has been successfully used for the quality control of enriched, cultured, and gene-modified NHPCD34+hematopoietic stem and progenitor cells (HSPCs) as well as sort-purified CD34 subsets for transplantation in the pig-tailed, cynomolgus, and rhesus macaque. It further allows the longitudinal assessment of primary specimens taken during the long-term follow-up post-transplantation in order to monitor homing, engraftment, and reconstitution of the bone marrow stem cell compartment.