Understanding the freezing responses of T cells and other subsets of human peripheral blood mononuclear cells using DSMO-free cryoprotectants.
Understanding the freezing responses of T cells and other subsets of human peripheral blood mononuclear cells using DSMO-free cryoprotectants.
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DOI：
10.1016/j.jcyt.2020.01.013
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发表时间：
2020-05
期刊：
影响因子：
4.5
通讯作者：
Hubel A
中科院分区：
文献类型：
--
作者：
Pi CH;Hornberger K;Dosa P;Hubel A
本研究检测了外周血单核细胞（PBMCs）和其中含有的特定白细胞亚群在由糖、糖醇和氨基酸组成的三种渗透液中冷冻保存时的冷冻反应。采用多目标差分进化算法优化冷冻保护剂组成，以同时优化辅助t细胞和细胞毒性t细胞的解冻后恢复。使用差分进化算法筛选各种配方显示解冻后恢复超过80%的两个t细胞亚群。流式细胞术对PBMC亚群的表型和活力进行了表征。解冻后辅助性t细胞和细胞毒性t细胞的恢复有显著差异。使用统计模型来分析个体渗透细胞的重要性以及解冻后恢复的三种t细胞亚群（包括辅助性t细胞、细胞毒性t细胞和自然杀伤t细胞）之间的相互作用。统计模型表明，这三个亚群的渗透物首选浓度水平和相互作用模式不同。pbmc解冻后培养72小时，以测定细胞的稳定性。由于解冻后淋巴细胞的凋亡是一个重要的问题，我们在解冻后立即和解冻后24小时分析了在无dmso冷冻保护剂中冷冻的辅助性t细胞和细胞毒性t细胞的凋亡情况。解冻后24小时，两种细胞类型的细胞活力均较解冻后立即下降。辅助性t细胞活力下降17%，细胞毒性t细胞活力下降10%。解冻后，两种细胞均有近30%的细胞处于早期凋亡状态，但24h后早期凋亡的细胞数量下降至20%以下。这项研究帮助我们确定了不同人类PBMC亚群的冷冻反应，使用渗透物的组合。
This study examined the freezing responses of peripheral blood mononuclear cells (PBMCs) and specific white blood cell subsets contained therein when cryopreserved in three combinations of osmolytes composed of sugars, sugar alcohols and amino acids. A differential evolution algorithm with multiple objectives was used to optimize cryoprotectant composition in order to optimize the post-thaw recoveries for both helper and cytotoxicity T-cells simultaneously. The screening of various formulations using differential evolution algorithm showed post-thaw recoveries over 80% for the two subsets of T-cells. The phenotypes and viabilities of PBMC subsets were characterized using flow cytometry. Significant differences between the post-thaw recovery for helper T-cell and cytotoxic T-cell were observed. Statistical models were used to analyze the importance of individual osmolytes and interactions between post-thaw recoveries of three subsets of T-cell including helper T-cells, cytotoxic T-cells and natural killer T-cells. The statistical model indicated that the preferred concentration levels of osmolytes and interaction modes were distinct between these three subsets studied. PBMCs were cultured for 72 hours post-thaw to determine the stability of the cells. As post-thaw apoptosis is a significant concern for lymphocytes, apoptosis of helper T-cell and cytotoxic T-cells frozen in a DMSO-free cryoprotectant was analyzed immediately post-thaw and 24 hours post-thaw. Both cell types showed a decrease in cell viability 24 hours post-thaw compared to immediately post-thaw. Helper T-cell viability dropped 17%, and cytotoxic T-cells had a 10% drop in viability. Immediately post-thaw, both cell types had >30% of cells in early apoptosis, but after 24 hours the number of cells in early apoptosis decreased to below 20%. This study helped us to identify the freezing responses of different human PBMC subsets using combinations of osmolytes.
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DOI：
10.3390/molecules22111789
发表时间：
2017-11-10
期刊：
Molecules (Basel, Switzerland)
影响因子：
--
作者：
de Abreu Costa L;Henrique Fernandes Ottoni M;Dos Santos MG;Meireles AB;Gomes de Almeida V;de Fátima Pereira W;Alves de Avelar-Freitas B;Eustáquio Alvim Brito-Melo G
通讯作者：
Eustáquio Alvim Brito-Melo G
影响因子：
2.2
作者：
Mata MM;Mahmood F;Sowell RT;Baum LL
通讯作者：
Baum LL
影响因子：
2.7
作者：
Luo, Ying;Wang, Peng;Gao, Yingtang
通讯作者：
Gao, Yingtang
影响因子：
3.8
作者：
Pi, Chia-Hsing;Yu, Guanglin;Hubel, Allison
通讯作者：
Hubel, Allison
影响因子：
2.7
作者：
Germann, Anja;Oh, Young-Joo;von Briesen, Hagen
通讯作者：
von Briesen, Hagen