Schroeder M L, Storb R, Goselink H, Gluckman E
Transplantation. 1977 Jan;23(1):33-8. doi: 10.1097/00007890-197701000-00006.
A satisfactory and reproducible technique of cryopreservation of canine lymphocytes for use in mixed leukocyte culture (MLC) and cell-mediated lympholysis tests has been developed. Dimethyl sulfoxide was used as the cryopreservation agent. Cells were frozen to -50 C at a controlled rate (-1 C/min) and stored at -169 C. The best preservation was obtained with a concentration of 10 x 10(6) lymphocytes/ml in Waymouth's MB-752/1 medium supplemented with 30% dog serum (MB-30). Before use in MLC or cell-mediated lympholysis tests, lymphocytes were rapidly thawed at 40 C and then rapidly diluted with MB-30 at room temperature. For best responses in MLC, one fresh component (either stimulating or stimulated cells or serum) was needed. The magnitudes of responses of frozen lymphocytes to phytohemagglutinin or in MLC were similar to those seen with fresh cells, but peak responses occurred usually 24 hr after those seen with fresh cells.
已开发出一种用于混合白细胞培养（MLC）和细胞介导的淋巴细胞溶解试验的犬淋巴细胞冷冻保存技术，该技术令人满意且可重复。使用二甲基亚砜作为冷冻保存剂。细胞以可控速率（-1℃/分钟）冷冻至-50℃，并储存在-169℃。在补充有30%犬血清（MB-30）的Waymouth's MB-752/1培养基中，以10×10⁶淋巴细胞/毫升的浓度可获得最佳保存效果。在用于MLC或细胞介导的淋巴细胞溶解试验之前，淋巴细胞在40℃快速解冻，然后在室温下用MB-30快速稀释。为了在MLC中获得最佳反应，需要一种新鲜成分（无论是刺激细胞还是被刺激细胞或血清）。冷冻淋巴细胞对植物血凝素或在MLC中的反应强度与新鲜细胞相似，但峰值反应通常比新鲜细胞晚24小时出现。
