梅花鹿鹿茸真皮层细胞的分离培养及冷冻保存  

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

作　　者：王守堂[1] 张璐[1] 田学超[1] 韩玉帅[1] 李当当[1] 杨德才[1] 郭斌[1] 岳占碰[1] 

机构地区：[1]吉林大学畜牧兽医学院,长春130062

出　　处：《中国农学通报》2012年第2期15-18,共4页Chinese Agricultural Science Bulletin

基　　金：高等学校博士学科点专项科研基金资助项目"甲状旁腺素相关肽及其受体在梅花鹿茸角内的表达及与茸角再生的关系"(20100061110078);国家自然科学基金资助项目"PTHrP-IHH信号通路对梅花鹿茸角再生的调控"(31072099);国家高技术研究发展计划资助项目"梅花鹿鹿茸再生相关基因的筛选;克隆及功能分析"(2007AA10Z150)

摘　　要：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5天可长至汇合。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。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.

关 键 词：梅花鹿鹿茸 真皮层细胞 体外培养 冷冻保存 

分 类 号：S8[农业科学—畜牧兽医] Q24[生物学—细胞生物学]