Document Detail

Effects of isolation and culture of turkey primary follicular oocytes on morphology and germinal vesicle integrity.
MedLine Citation:
PMID:  10734429     Owner:  NLM     Status:  MEDLINE    
A novel approach to the production of transgenic poultry is to use primary follicular oocytes (PFOs). However, fundamental information regarding the impact of isolation and culture procedures on PFO integrity is lacking. This study describes the isolation and culture of PFOs from mature turkeys and the effects of these procedures on PFO morphology and germinal vesicle (GV) integrity. To isolate PFOs, ovarian cortex was incubated in trypsin-EDTA alone or further incubated in collagenase plus hyaluronidase (CH). About 200 to 500 PFOs, ranging in size from less than 100 microns in diameter to 1,000 microns, were recovered from each ovary. The culture of PFOs less than 100 microns in diameter for 4 h resulted in blebbing of the oolemma followed by extrusion of ooplasm. Primary follicular oocytes 100 to 250 microns in diameter survived culture for 24 h whereas larger PFOs survived for up to 7 d. Those PFOs with intact granulosa cell investments survived longer than those fully or partially denuded of granulosa cells with CH. Co-culture of PFOs (100 to 250 microns in diameter) on a monolayer of granulosa cells derived from mature, yellow-yolk follicles augmented PFO survival rates. The rate of GV breakdown was not influenced by the isolation or culture of the PFO. These data provide the basis for developing procedures for the in vitro maturation and in vitro fertilization of isolated PFOs.
M R Bakst; D Gliedt; V Akuffo; W Potts; S K Gupta
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Publication Detail:
Type:  Journal Article    
Journal Detail:
Title:  Theriogenology     Volume:  50     ISSN:  0093-691X     ISO Abbreviation:  Theriogenology     Publication Date:  1998 Nov 
Date Detail:
Created Date:  2000-04-07     Completed Date:  2000-04-07     Revised Date:  2004-11-17    
Medline Journal Info:
Nlm Unique ID:  0421510     Medline TA:  Theriogenology     Country:  UNITED STATES    
Other Details:
Languages:  eng     Pagination:  1121-30     Citation Subset:  IM    
Germplasm and Gamete Physiology Laboratory, U.S. Department of Agriculture, Beltsville, MD 20705, USA.
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MeSH Terms
Cell Size
Cells, Cultured
Oocytes / cytology*,  physiology*
Ovarian Follicle / cytology*

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