Yu Fang-Jian, Zeng Chang-Jun, Zhang Yan, Wang Cheng-Dong, Xiong Tie-Yi, Fang Sheng-Guo, Zhang He-Min
1College of Life Sciences, Zhejiang University, Hangzhou, China.
2Farm Animal Genetic Resources Exploration and Innovation Key Laboratory of Sichuan Province, Sichuan Agricultural University, Ya'an, Sichuan Province, China.
Biopreserv Biobank. 2015 Jun;13(3):195-9. doi: 10.1089/bio.2014.0073.
The giant panda Ailuropoda melanoleuca is an endangered species and is a symbol for wildlife conservation. Although efforts have been made to protect this rare and endangered species through breeding and conservative biology, the long-term preservation of giant panda genome resources (gametes, tissues, organs, genomic libraries, etc.) is still a practical option. In this study, the giant panda skeletal muscle-derived cell line was successfully established via primary explants culture and cryopreservation techniques. The population doubling time of giant panda skeletal cells was approximately 33.8 h, and this population maintained a high cell viability before and after cryopreservation (95.6% and 90.7%, respectively). The two skeletal muscle-specific genes SMYD1 and MYF6 were expressed and detected by RT-PCR in the giant panda skeletal muscle-derived cell line. Karyotyping analysis revealed that the frequencies of giant panda skeletal muscle cells showing a chromosome number of 2n=42 ranged from 90.6∼94.2%. Thus, the giant panda skeletal muscle-derived cell line provides a vital resource and material platform for further studies and is likely to be useful for the protection of this rare and endangered species.
大熊猫（Ailuropoda melanoleuca）是一种濒危物种，也是野生动物保护的象征。尽管已通过繁殖和保护生物学等手段努力保护这一珍稀濒危物种，但长期保存大熊猫基因组资源（配子、组织、器官、基因组文库等）仍是一种切实可行的选择。在本研究中，通过原代外植体培养和冷冻保存技术成功建立了大熊猫骨骼肌来源的细胞系。大熊猫骨骼肌细胞的群体倍增时间约为33.8小时，该群体在冷冻保存前后均保持较高的细胞活力（分别为95.6%和90.7%）。通过RT-PCR在大熊猫骨骼肌来源的细胞系中表达并检测到了两个骨骼肌特异性基因SMYD1和MYF6。核型分析显示，染色体数目为2n = 42的大熊猫骨骼肌细胞频率在90.6%至94.2%之间。因此，大熊猫骨骼肌来源的细胞系为进一步研究提供了重要资源和物质平台，可能有助于保护这一珍稀濒危物种。
