An Investigation of Factors Affecting Cryopreservation of Murine Splenic Lymphocytes
- VernacularTitle:深低温保存小鼠脾淋巴细胞的影响因素
- Author:
Jianming CAI
- Publication Type:Journal Article
- Keywords:
cryoprotective agent;
preservation;
lymphocyte;
graft vs host disease;
spleen;
mice
- From:
Academic Journal of Second Military Medical University
1981;0(03):-
- CountryChina
- Language:Chinese
-
Abstract:
The main factors which affect the cryopreservation of munne splenic lymphocytes were studied. The results showed that DMSO had some toxic effects on lymphocytes when their concentration was more than 10%. However, when their concentration was 10%, they could effectively protect the cells from freezing injuries and had not significant toxicity. The cooling rate giving optimal survival was 1℃/min. Compared with the lymphocytes sealed in HF bags, the functional recovery of fast frozen (6℃/min)lymphocytes sealed in plastic tubes decreased more severely. Diluting the thawed lymphocytes slowly or at 25-30℃gave more greatly improved recovery than rapidly or at ice bath. Lysing RBC by Tris-NH4C1 might increase the susceptibility of splenic lymphocytes from some strain mice to cryoinjuries. Using selected optimal conditions, the viability tested by Trypan Blue Dye Exclusion and the functions measured by [3H] -TdR incorporation of splenic lymphocytes in response to Con A, LPS and MLR in vitro or the ability to initiate GVHD in vivo were well cryopreserved. The time of storage at-196℃ did not affect the function of lymphocytes in this report.
