梅花鹿鹿茸真皮层细胞的分离培养及冷冻保存

为了研究梅花鹿鹿茸真皮层细胞的生物学特性，取梅花鹿鹿茸生长顶端的真皮层组织，分离真皮层细胞进行体外培养及冷冻保存。结果表明，在含10 %胎牛血清(FBS)的DMEM培养基中，鹿茸真皮层细胞能进行短期体外培养，培养的细胞呈长梭形或菱形，细胞的生长状况良好，传代培养的细胞培养5d可长至汇合。传4代以前的细胞形态均一，边缘光滑，胞质均匀透明，细胞排列紧密；传5代后，细胞伸展变为扁平形，细胞突起增多，边缘不光滑，汇合后细胞排列疏松。以含5%二甲基亚砜(DMSO)和10%FBS的DMEM为冻存液，经梯度降温后冻存，真皮层细胞复苏后的存活率较高。

Culture and Cryopreservation of Sika Deer Antler Dermis Layer Cells in Vitro

To study the biological characteristics of sika deer antler dermis layer cells, the dermis layer of growing tip in sika deer antler was collected and a culture and cryopreservation system in vitro for the dermis layer ceils was established. The results showed that the antler dermis layer ceils could be cuhured well within a short term in DMEM medium with 10% FBS in vitro. The cultured dermis layer cells were spindle or rhombic and the cultured ceils merged each other after 5 days. The cells before the fourth generation appeared uniform. The edge of the cells was smooth and the cytoplasm was uniform and transparent. The cultured cells were closely arranged. The cultured cells changed into flat after the fifth generation and extended more apophysises. The edge of the cultured cells was not smooth and the cells were loosely arranged after merged. The suitable cryopreserved condition for the dermis layer cells was DMEM medium containing 5%DMSO and 10%FBS with gradient-cooling.