Law P, Alsop P, Dooley D C, Meryman H T
Cryobiology. 1983 Dec;20(6):644-51. doi: 10.1016/0011-2240(83)90068-8.
The feasibility of using hypo- or hypertonic stress to selectively destroy lymphocytes while sparing stem cells was investigated. Lymphocytes were isolated from peripheral blood and exposed to Hanks' balanced salt solutions ranging in concentration from 66 to 2700 mOsm. The Boyle-van't Hoff plot of cell volume versus reciprocal osmolality was linear. Following osmotic stress, viabilities of the lymphocytes and the granulocyte-monocyte progenitor cells (CFUc) were determined. Lymphocyte viability was assessed by tritiated thymidine incorporation following mitogen stimulation. CFUc viability was measured with the soft agar colony assay. Both types of cells were found to possess high osmotic tolerances compared to other blood cells. While progenitor cells in general appeared to survive anisotonic exposure somewhat better than lymphocytes, significant statistical differences were not established for most situations. The highest degree of CFUc enrichment was twofold, but there was a concomitant 50% drop in CFUc survival. These results suggest that osmotic stress is not a useful procedure for the separation of peripheral blood lymphocytes and stem cells.
研究了利用低渗或高渗应激选择性破坏淋巴细胞同时保留干细胞的可行性。从外周血中分离淋巴细胞，并将其暴露于浓度范围为66至2700 mOsm的汉克斯平衡盐溶液中。细胞体积与渗透压倒数的博伊尔-范特霍夫图呈线性关系。在渗透应激后，测定淋巴细胞和粒细胞-单核细胞祖细胞（CFUc）的活力。通过有丝分裂原刺激后掺入氚标记的胸腺嘧啶核苷来评估淋巴细胞活力。用软琼脂集落试验测量CFUc活力。与其他血细胞相比，发现这两种类型的细胞都具有较高的渗透压耐受性。虽然一般来说祖细胞似乎在不等渗暴露下比淋巴细胞存活得稍好一些，但在大多数情况下没有建立显著的统计学差异。CFUc的最高富集程度为两倍，但同时CFUc存活率下降了50%。这些结果表明，渗透应激不是分离外周血淋巴细胞和干细胞的有用方法。
