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

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

Research output: Contribution to journalArticle

7 Scopus citations

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.

Original languageEnglish (US)
Pages (from-to)547-556
Number of pages10
JournalJournal of Assisted Reproduction and Genetics
Volume29
Issue number6
DOIs
StatePublished - Jun 1 2012

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Keywords

  • Follicular fluid
  • Human granulosa cell
  • Immunomagnetic beads
  • In vitro fertilization
  • MIS receptor
  • Purification
  • Red blood cell contamination

ASJC Scopus subject areas

  • Reproductive Medicine
  • Genetics
  • Obstetrics and Gynecology
  • Developmental Biology
  • Genetics(clinical)

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