Cryobiophysical characteristics of genetically modified hematopoietic progenitor cells
Cryobiophysical characteristics of genetically modified hematopoietic progenitor cells
复制标题
DOI：
10.1006/cryo.1999.2157
复制
发表时间：
1999-03-01
期刊：
影响因子：
2.7
通讯作者：
Darr, TB
中科院分区：
文献类型：
--
作者：
Hubel, A;Norman, J;Darr, TB
使用冷冻显微镜观察正常供体和粘多糖病I型供体的造血祖细胞的冷冻反应，并使用水运输和细胞内冰形成的理论模型进行分析。本研究中使用的来自患有MPS I的供体的细胞被培养并用用于人类基因治疗临床前研究的α - l -伊杜糖醛酸酶（IDUA）酶的逆转录病毒载体转导。在培养和转导过程中，对表达CD34抗原的造血祖细胞在不同时间点的水转运和细胞内冰形成（IIF）特征进行了测定。不同来源（正常供体与MPST供体）和不同培养条件（新鲜分离与培养和转导）的细胞在水输送、渗透性失活细胞体积分数和通透性方面有统计学意义的变化。其中，L-pg和E-a在离体培养后增加，仅在培养3天后就观察到通透性参数的变化。同样，造血祖细胞的IIF特性也会受到培养和转导过程的影响。从患有MPS的供体中新鲜分离的细胞的LIF特征在统计上与来自同一供体的培养和转导细胞的LIF特征不同。为了治疗目的而体外培养的细胞的低温保存能力将需要了解由培养条件引起的生物物理变化以及这些变化影响生存能力的方式。(C) 1999学术出版社。
The freezing responses of hematopoietic progenitor cells isolated from normal donors and from donors with mucopolysaccharidosis type I(MPS I) were determined using cryomicroscopy and analyzed using theoretical models for water transport and intracellular ice formation. The cells from donors with MPS I used in this investigation were cultured and transduced with a retroviral vector for the alpha-L-iduronidase (IDUA) enzyme in preclinical studies for human gene therapy. The water transport and intracellular ice formation (IIF) characteristics were determined at different time points in the culture and transduction process for hematopoietic progenitor cells expressing CD34 antigen from donors with MPS I and from normal donors. There were statistically significant changes in water transport, osmotically inactive cell volume fraction, and permeability between cells from different sources (normal donors vs donors with MPST) and different culture conditions (freshly isolated vs cultured and transduced). Specifically, L-pg and E-a increased after ex vivo culture of the cells and the changes in permeability parameters were observed after as little as 3 days in culture. Similarly, the IIF characteristics of hematopoietic progenitor cells can also be influenced by the culture and transduction process. The LIF characteristics of freshly isolated cells from donors with MPS were statistically distinct from those of cultured and transduced cells from the same donor, The ability to cryopreserve cells which are cultured ex vivo for therapeutic purposes will require an understanding of the biophysical changes resulting from the culture conditions and the manner in which these changes influence viability. (C) 1999 Academic Press.