Document Detail

Advanced methods of isolation and identification of porcine primordial follicles.
MedLine Citation:
PMID:  17250982     Owner:  NLM     Status:  MEDLINE    
An effective isolation and identification method of primordial follicles would greatly benefit the animal production practice, transgenic animal production and endangered species conservation in the future. This study has not only advanced the isolation method but also developed an identification marker of primordial follicles. After enzymatic digestion, Percoll gradient centrifugation and mesh filtrations, the obtained follicular separations were then subjected to a cell sorter in order to collect primordial follicles. The study greatly improved the yield of primordial follicles (from about 1.85 x 10(5) to 7.79 x 10(5) per prepubertal ovary) by means of increasing cell layer number from 1 ml to 2.5 ml after Percoll gradient centrifugation. Based on traditional morphological criteria, the purity of recovered primordial follicles was averagely about 82.43+/-9.41% (n=5) because of their similar size and appearance with somatic cells. To further exactly appreciate the purity of sorted primordial follicles, a germ cell-specific protein, MSY2, was used to recognize the oocytes of primordial follicles. The results of repeat experiments showed that about 98.31+/-0.73% (n=4) of the primordial follicles cluster was MSY2-positive, which indicated the identification method of primordial follicles was more effective and high-yielded than the previous methods because of its higher purity.
Kerong Shi; Xuefeng Yuan; Liying Du; Dengke Pan; Yunhai Zhang; Yonghui Zhao; Xuemei Deng; Xiaoxiang Hu; Changxin Wu; Ning Li
Publication Detail:
Type:  Evaluation Studies; Journal Article; Research Support, Non-U.S. Gov't     Date:  2006-11-26
Journal Detail:
Title:  Animal reproduction science     Volume:  101     ISSN:  0378-4320     ISO Abbreviation:  Anim. Reprod. Sci.     Publication Date:  2007 Sep 
Date Detail:
Created Date:  2007-06-29     Completed Date:  2007-10-04     Revised Date:  -    
Medline Journal Info:
Nlm Unique ID:  7807205     Medline TA:  Anim Reprod Sci     Country:  Netherlands    
Other Details:
Languages:  eng     Pagination:  163-71     Citation Subset:  IM    
State Key Laboratory for Agrobiotechnology, College of Biological Science, China Agricultural University, No. 2 Yuan-Ming-Yuan West Road, Haidian District, Beijing 100094, China.
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MeSH Terms
Cell Count
Cell Separation
Cell Survival
Flow Cytometry
Ovarian Follicle / cytology*
Staining and Labeling / methods
Tissue and Organ Harvesting / methods*

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