移植瘤株冷冻保护的实验研究

本文介绍了人肝癌细胞析细胞及其移植瘤组织经冻存，复苏后检测琥珀酸脱氢酶（ＳＤＥ）活性，成瘤率和电镜检查，选择１０％ＤＭＳＯ或１０％甘油作为冷冻保护剂来冻存细胞，它们先在冰箱里缓慢降温冷却（－０．８℃／ｍｉｎ），后在液氮罐口的液氮气中冷冻（－１０℃／ｍｉｎ）１天，最后置入液氮内，冷并保护后的冻存或组织复苏后显示高的ＳＤＥ活性，１０％ＤＭＳＯ冻存的细胞ＳＤＥ活性比１０％甘油者高，两者相比有显著差别（Ｐ

Experiment of preserving efficiency of xenografts freezed in liquid nitrogen

It has not been done into detail that whether human Xenografted tumors could be frozen in liquid nitrogen(- 196 ),thawed and recovered. A human hepatocellular carcinoma cell line (H91O1 ) and its xenografts were tested by succino dehydrogenase (SDE)activity (MTT test) after freezing and thawing and being studied by electron microscope.Two preservative liquids included lO% DMSo or 1o% glyceroI were adopted for the freezing in which the cells or tissues were controlled to be reduced to a minimum rate of cooling (-0.8t / min) in the gas of liquid nitrogen for 24 hours and programmed in liquid nitrogen.It was shown that H9lOl cells or tissues after freezing and thawing presented a high SDE activity (72%~ 93%). The difference of SDE activity between the cells or tissues preserved in 10% DMSO and that preserved in lO% blycerol were significant (P< 0.01 ). The inoculating efficiency of xenografts stored in 10% DMSO was same to that stored in lO% glycerol. Observation by electron microscoPe, tumor cells frozen by both of the preservative medium showed more"black"cell and less"light"cells. Both"black"and"light"ce1ls showed an integral structure of mitochondria membrane and cytoplasmic membrane. Although there was not a general acceptance of the claims that DMSO was preferable to glycerol as an agent for pre venting different organism from freezing damages, it was satisfactory that xenografts pre served in 10% DMSO could improve the survival rates of tumor cells frozen in liquid nitrogen