Clinical mesenchymal stromal cell products undergo functional changes in response to freezing.
Clinical mesenchymal stromal cell products undergo functional changes in response to freezing.
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DOI：
10.1016/j.jcyt.2014.06.008
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发表时间：
2015-01
期刊：
影响因子：
4.5
通讯作者：
Hubel, Allison
中科院分区：
文献类型：
--
作者：
Pollock, Kathryn;Sumstad, Darin;Kadidlo, Diane;McKenna, David H.;Hubel, Allison
目前的MSC冷冻保存方法导致不同的解冻后恢复和冷冻引起的表型变化。本研究的目的是确定离体细胞扩增对MSC表型的影响以及所得表型对冷冻和解冻的反应。人骨髓穿刺液购自Lonza（Walkersville，MD）。分离MSC，并在离体培养中在6代（8-10倍/代）内评估细胞的总计数、活力、凋亡和衰老。将每次传代时收获的一半细胞重新铺板以继续培养，另一半细胞以1°C/min的速率在受控速率冷冻机中冷冻。将冷冻样品储存在液氮中，解冻，并在解冻后立即和48小时重新评估总细胞计数、活力和衰老。冷冻前或解冻后样品之间的活力无显著差异。在冷冻前培养物中衰老随时间增加，并且在冷冻前和解冻后经历生长停滞的一个样品中显著更高。冷冻导致所有样品中相似的初始解冻后恢复，但仅在高度衰老的样品中观察到解冻后48小时生长停滞。高冷冻衰老似乎与MSC样品中解冻后功能差相关，但需要进行额外的研究以获得足够大的样本量来量化结果。
Current methods of MSC cryopreservation result in variable post thaw recovery and phenotypic changes due to freezing. The objective of this investigation is to determine the influence of ex-vivo cell expansion on phenotype of MSCs and the response of resulting phenotypes to freezing and thawing. Human bone marrow aspirate was purchased from Lonza (Walkersville, MD). MSCs were isolated, and cells were assessed for total count, viability, apoptosis, and senescence over 6 passages (8–10 doublings/passage) in ex vivo culture. One half of cells harvested at each passage were re-plated for continued culture, and the other half were frozen at 1°C/min in a controlled rate freezer. Frozen samples were stored in liquid nitrogen, thawed, and reassessed for total cell count, viability, and senescence immediately and 48 hours post thaw. Viability did not differ significantly between samples pre freeze or post thaw. Senescence increased over time in pre freeze culture, and was significantly higher in one sample that experienced growth arrest both pre freeze and post thaw. Freezing resulted in similar initial post thaw recovery in all samples, but 48 hour post thaw growth arrest was observed in the sample with high senescence only. High freeze senescence appears to correlate with poor post thaw function in MSC samples, but additional studies are necessary to obtain a sample size large enough to quantify results.
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影响因子：
3.7
作者：
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