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Journal of Assisted Reproduction and Genetics
Published in: Journal of Assisted Reproduction and Genetics 6/2012

01-06-2012 | Technological Innovations

Development of a novel protocol for isolation and purification of human granulosa cells

Authors: R. A. Chilvers, Y. H. Bodenburg, L. A. Denner, R. J. Urban

Published in: Journal of Assisted Reproduction and Genetics | Issue 6/2012

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Abstract

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.
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Metadata
Title
Development of a novel protocol for isolation and purification of human granulosa cells
Authors
R. A. Chilvers
Y. H. Bodenburg
L. A. Denner
R. J. Urban
Publication date
01-06-2012
Publisher
Springer US
Published in
Journal of Assisted Reproduction and Genetics / Issue 6/2012
Print ISSN: 1058-0468
Electronic ISSN: 1573-7330
DOI
https://doi.org/10.1007/s10815-012-9739-5

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