Vaccine & Gene Therapy Institute, Oregon Health & Science University, Beaverton, Oregon, USA.
Oregon National Primate Research Center, Oregon Health & Science University, Beaverton, Oregon, USA.
J Clin Apher. 2021 Feb;36(1):67-77. doi: 10.1002/jca.21842. Epub 2020 Sep 17.
Macaques are physiologically relevant animal models of human immunology and infectious disease that have provided key insights and advanced clinical treatment in transplantation, vaccinology, and HIV/AIDS. However, the small size of macaques is a stumbling block for studies requiring large numbers of cells, such as hematopoietic stem cells (HSCs) for transplantation, antigen-specific lymphocytes for in-depth immunological analysis, and latently-infected CD4+ T-cells for HIV cure studies. Here, we provide a detailed protocol for collection of large numbers of HSCs and T-cells from cynomolgus macaques as small as 3 kg using the Terumo Spectra Optia apheresis system, yielding an average of 5.0 × 10 total nucleated cells from mobilized animals and 1.2 × 10 total nucleated cells from nonmobilized animals per procedure. This report provides sufficient detail to adapt this apheresis technique at other institutions, which will facilitate more efficient and detailed analysis of HSCs and their progeny blood cells.
猕猴是研究人类免疫学和传染病的重要生理相关动物模型,为移植、疫苗学和 HIV/AIDS 领域的临床治疗提供了关键的见解。然而,猕猴体型较小,对于需要大量细胞的研究构成了障碍,例如移植所需的造血干细胞(HSCs)、用于深入免疫分析的抗原特异性淋巴细胞,以及用于 HIV 治愈研究的潜伏感染 CD4+ T 细胞。在这里,我们提供了一个详细的方案,使用 Terumo Spectra Optia 血细胞分离机从体重仅 3 公斤的食蟹猴中采集大量的 HSCs 和 T 细胞,从动员的动物中平均获得 5.0×10^6 个总核细胞,从非动员的动物中平均获得 1.2×10^6 个总核细胞。本报告提供了足够的详细信息,可使其他机构采用这种细胞分离技术,从而更有效地对 HSCs 及其祖细胞进行详细分析。