Document Detail


Fibroblasts from the new-born male testicle of Guangxi Bama mini-pig ( Sus scrofa) can support nuclear transferred embryo development in vitro.
MedLine Citation:
PMID:  19224667     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
Miniature pigs are valuable for research in xenotransplantation and as models for investigating human diseases. Although many mammalian species have been cloned, the success rates have been very low, especially in the pig. In the present study, an attempt was made to optimize somatic cell nuclear transfer (SCNT) protocols for use in the production of the Guangxi Bama mini-pig. Firstly, mini-pig fibroblast cells from a new-born Guangxi Bama piglet were isolated and cultured. Cell type was identified by fluorescence immunocytochemistry (ICC); the cells expressed cimentin, but not cytoceratin and follicular stimulation hormone receptor (FSHR). Secondly, the optimal cell cycle synchronization protocol for treating fibroblast cells from the newborn piglet's testicle was investigated by contact inhibition and serum starvation. When fibroblast cells were treated by contact inhibition, a higher fusion (66.0% vs. 58.3%, p > 0.05) and blastocyst production (20.8% vs. 15.1, p > 0.05) rates were obtained than with serum starvation. Thirdly, to examine the ability of old cells to be morphologically remodelled after activation, testicular fibroblasts (passage 10-14) were introduced into enucleated oocytes; enlarged nuclei were formed in most of the reconstructed embryos at 6 h and enlarged nuclei or distinct pseudopronuclei were formed in nearly all the reconstructed embryos at 12 h. The old donor cell could be morphologically remodelled correctly and was competent to support embryo development to the blastocyst in vitro. Fourthly, the in vitro development potential of the cloned embryos was investigated using two types of donor cell: ear fibroblasts and low or high passage testicular fibroblasts. The rate of fusion was highest using low passage testicle fibroblasts (84.5% vs. 69.8% and 80.0%, p < 0.05), as was development to the blastocyst stage (14.6% vs. 7.7% and 6.3%, p < 0.05). Finally, the effect of phytohaemagglutinin (PHA) on parthenogenetic and cloned embryo development was examined. The PHA had no significant effect on the parthenogenetic embryos, but cloned embryo development to the blastocyst stage was significantly increased by PHA (10 microg/ml), (13.4% vs. 5.6% and 5.6%, p < 0.05).
Authors:
Hong-Bo Liu; Pei-Ru Lv; Xiao-Gan Yang; Xiao-E Qin; Dao-Yuan Pi; Yang-Qing Lu; Ke-Huan Lu; Sheng-Sheng Lu; Dong-Sheng Li
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't     Date:  2009-02-19
Journal Detail:
Title:  Zygote (Cambridge, England)     Volume:  17     ISSN:  0967-1994     ISO Abbreviation:  Zygote     Publication Date:  2009 May 
Date Detail:
Created Date:  2009-04-01     Completed Date:  2009-06-12     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  9309124     Medline TA:  Zygote     Country:  England    
Other Details:
Languages:  eng     Pagination:  147-56     Citation Subset:  IM    
Affiliation:
Guangxi Key Laboratory of Subtropical Bio-Resource Conservation and Utilization, Guangxi University, Nanning, Guangxi 530004, China.
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MeSH Terms
Descriptor/Qualifier:
Animals
Blastocyst / physiology*
Cell Fusion
Embryonic Development / drug effects,  physiology
Fibroblasts / cytology,  physiology*
Male
Nuclear Transfer Techniques
Oocytes / physiology
Parthenogenesis / drug effects,  physiology*
Phytohemagglutinins / pharmacology
Swine
Swine, Miniature*
Testis / cytology*
Vimentin / metabolism
Chemical
Reg. No./Substance:
0/Phytohemagglutinins; 0/Vimentin

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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