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Fibroblast cell line establishment, cryopreservation and interspecies embryos reconstruction in red panda (Ailurus fulgens)

Published online by Cambridge University Press:  01 May 2009

Yong Tao ,
Jianming Liu ,
Yunhai Zhang ,
Meiling Zhang ,
Junshun Fang ,
Wei Han ,
Zhizhong Zhang ,
Ya Liu ,
Jianping Ding  and
Xiaorong Zhang
Yong Tao
Affiliation:
College of Animal Science and Technology, Anhui Agricultural University, Hefei, 230036, China. These authors contributed equally to the paper.
Jianming Liu
Affiliation:
College of Animal Science and Technology, Anhui Agricultural University, Hefei, 230036, China. These authors contributed equally to the paper.
Yunhai Zhang
Affiliation:
College of Animal Science and Technology, Anhui Agricultural University, Hefei, 230036, China.
Meiling Zhang
Affiliation:
College of Animal Science and Technology, Anhui Agricultural University, Hefei, 230036, China.
Junshun Fang
Affiliation:
College of Animal Science and Technology, Anhui Agricultural University, Hefei, 230036, China.
Wei Han
Affiliation:
College of Animal Science and Technology, Anhui Agricultural University, Hefei, 230036, China.
Zhizhong Zhang
Affiliation:
Hefei Wild Animal Park, Hefei, 230061, Anhui Province, China.
Ya Liu
Affiliation:
College of Animal Science and Technology, Anhui Agricultural University, Hefei, 230036, China.
Jianping Ding
Affiliation:
College of Animal Science and Technology, Anhui Agricultural University, Hefei, 230036, China.
Xiaorong Zhang*
Affiliation:
College of Animal Science and Technology, Anhui Agricultural University, Changjiang West Road 130, Hefei, 230036, China.
*
All correspondence to: Zhang Xiaorong. College of Animal Science and Technology, Anhui Agricultural University, Changjiang West Road 130, Hefei, 230036, China. Tel: +8 6551 5782 488. Fax: +8 6551 5785 543. e-mail: zxr@ahau.edu.cn
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Summary

In evolution, the red panda (Ailurus fulgens) plays a pivotal role in the higher level phylogeny of arctoides carnivore mammals. The red panda inhabits certain Asian countries only and its numbers are decreasing. Therefore, the development of feasible ways to preserve this species is necessary. Genetic resource cryopreservation and somatic cell nuclear transfer (SCNT) have been used extensively to rescue this endangered species. The present study describes the establishment, for the first time, of a red panda ear fibroblast cell line, which was then cryopreserved, thawed and cultured. Through micromanipulation, interspecies embryos were reconstructed using the cryopreserved–thawed fibroblasts of the red panda as the donor and rabbit oocytes as recipients. A total of 194 enucleated rabbit oocytes were reconstructed with red panda ear fibroblasts; enucleated oocytes were activated without fusion as the control. The results show that the fibroblast cell line was established successfully by tissue culture and then cryopreserved in liquid nitrogen. Supplementation with 20% fetal bovine serum and 8% dimethyl sulphoxide in basic medium facilitated the cryopreservation. The interspecies embryos were successfully reconstructed. The cleavage, morulae and blastocyst rates after in vitro culture were 71, 47 and 23% (31/194), respectively. This study indicated that a somatic cell line could be established and cryopreserved from red panda and that rabbit cytoplast supports mitotic cleavage of the red panda karyoplasts and is capable of reprogramming the nucleus to achieve blastocysts.

Keywords

Ear fibroblastInterspecies somatic cell nuclear transferRabbitRed panda
Type
Research Article
Information
Zygote , Volume 17 , Issue 2 , May 2009 , pp. 117 - 124
Copyright
Copyright © Cambridge University Press 2009

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