Document Detail


Development of a novel protocol for isolation and purification of human granulosa cells.
MedLine Citation:
PMID:  22407611     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
PURPOSE: To develop an optimal method of isolation and purification of human granulosa cells from ovarian follicular fluid.
METHODS: Follicular fluid was collected from patients undergoing oocyte retrieval. A series of isolation and purification techniques was performed, involving density gradient centrifugation and use of different antibody-bead complexes.
RESULTS: The highest percent yield of live purified granulosa cells came from density gradient centrifugation using sucrose polymer followed by positive selection of granulosa cells using primary antibody to MISRII and secondary antibody coupled to iron oxide beads.
CONCLUSIONS: A novel protocol for granulosa cell purification has been developed yielding samples that are largely free of nondesirable cells. This protocol provides a purification solution, especially for patient samples that have significant RBC contamination.
Authors:
R A Chilvers; Y H Bodenburg; L A Denner; R J Urban
Publication Detail:
Type:  Journal Article; Research Support, N.I.H., Extramural     Date:  2012-03-10
Journal Detail:
Title:  Journal of assisted reproduction and genetics     Volume:  29     ISSN:  1573-7330     ISO Abbreviation:  J. Assist. Reprod. Genet.     Publication Date:  2012 Jun 
Date Detail:
Created Date:  2012-06-08     Completed Date:  2012-11-30     Revised Date:  2013-06-26    
Medline Journal Info:
Nlm Unique ID:  9206495     Medline TA:  J Assist Reprod Genet     Country:  Netherlands    
Other Details:
Languages:  eng     Pagination:  547-56     Citation Subset:  IM    
Affiliation:
Department of Obstetrics & Gynecology, Division of Reproductive Endocrinology and Infertility, University of Texas Medical Branch, Galveston, TX, USA. rachilvers@gmail.com
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MeSH Terms
Descriptor/Qualifier:
Antibodies, Monoclonal
Blotting, Western
Buffers
Carrier Proteins / immunology
Cell Separation / methods*
Centrifugation, Density Gradient
Cytological Techniques / methods*
Erythrocytes
Female
Follicular Fluid / cytology
Granulosa Cells*
Humans
Seminal Plasma Proteins / immunology
Grant Support
ID/Acronym/Agency:
NIH RO1HD044566-01A2/HD/NICHD NIH HHS
Chemical
Reg. No./Substance:
0/Antibodies, Monoclonal; 0/Buffers; 0/Carrier Proteins; 0/FSIP1 protein, human; 0/Seminal Plasma Proteins
Comments/Corrections

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