牙髓成纤维细胞传代培养与冻存复苏后生长情况比较

目的：比较牙髓成纤维细胞在体外传代培养及冻存复苏后细胞生长情况的差异,探讨其应用价值。方法：采用组织块培养法对牙髓组织标本进行体外成纤维细胞的原代及传代培养,观察成纤维细胞形态及生物学特性,选取第4代处于对数生长期的细胞梯度降温后置入液氮中（196℃）保存3个月,比较经传代培养细胞与复苏后成纤维细胞的生长情况,用MTT法分别绘制细胞生长曲线,比较细胞生长情况。结果：经冻存保存3个月后牙髓细胞复苏率超过80%,而细胞形态未见明显的改变。复苏后细胞生长较对照组慢,进入对数生长期的时间较对照组略长。但两者生长曲线基本一致。结论：牙髓成纤维细胞传代培养与冷冻复苏后形态及生长特性基本相同,冻存的成纤维细胞可以成功的复苏。

Study on Culture, Cryopreservation and Revivification of Fibroblasts from Dental Pulp Tissue

Objective： To investigate the difference of biological characteristics of dental pulp fibroblasts cultured in vitro after passage and after revivified from cryopreservation. Mtehods： The fibroblasts from dental pulp tissue were respectively cultured and subcultured in vitro. Cell morphology of fibroblasts was observed. Growth curve was used to evaluate the proliferation of cells. The fourth passage cells in logarithmic growth phase were placed in refrigerator for gradient cooling and then were preserved in liquid nitrogen（- 196 ℃ ） for 3 months. Then, cell morphology was observed. Results： The results indicated that the fibroblasts after cryopreservation and revivification exhibited similar morphology and growth curve, The primary cells from pulp tissue grew faster. The frozen cells were resuscitated successfully after 3 months, the resuscitated rates were more than 80 %, and their morphology was not obviously changed. But frozen cells requested more time to enter into the logarithmic growth phase. Coclusion： The fibroblasts isolated from pulp tissue exhibited similar morphology and growth characteristic . In vitro, its cryopereservation and resuscitation were successful.