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中国农学通报 ›› 2012, Vol. 28 ›› Issue (2): 15-18.doi: 10.11924/j.issn.1000-6850.2011-1862

所属专题: 生物技术

• 畜牧 动物医学 蚕 蜂 • 上一篇    下一篇

梅花鹿鹿茸真皮层细胞的分离培养及冷冻保存

王守堂 张璐 田学超 韩玉帅 李当当 杨德才 郭斌 岳占碰   

  • 收稿日期:2011-06-23 修回日期:2011-08-01 出版日期:2012-01-15 发布日期:2012-01-15
  • 基金资助:

    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;PTHrP-IHH信号通路对梅花鹿茸角再生的调控;梅花鹿鹿茸再生相关基因的筛选、克隆及功能分析

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

  • Received:2011-06-23 Revised:2011-08-01 Online:2012-01-15 Published:2012-01-15

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。

关键词: 马铃薯, 马铃薯, 未授粉子房培养, 离体培养, 双单体植株

Abstract:

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 cells was established. The results showed that the antler dermis layer cells could be cultured 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 cells 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.

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