title: Development of a novel protocol for isolation and purification of human granulosa cells.
abstract: OBJECTIVE
  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.affiliation: Department of Obstetrics & Gynecology, Division of Reproductive Endocrinology and Infertility, University of Texas Medical Branch, Galveston, TX, USA. rachilvers@gmail.com
authors.forename: R A
authors.initials: RA
authors.lastname: Chilvers
authors.forename: Y H
authors.initials: YH
authors.lastname: Bodenburg
authors.forename: L A
authors.initials: LA
authors.lastname: Denner
authors.forename: R J
authors.initials: RJ
authors.lastname: Urban
mesh_terms: D000911:Antibodies, Monoclonal; D015153:Blotting, Western; D002021:Buffers; D002352:Carrier Proteins / Q000276:immunology; D002469:Cell Separation / Q000379:methods*; D002499:Centrifugation, Density Gradient; D003584:Cytological Techniques / Q000379:methods*; D004912:Erythrocytes; D005260:Female; D015571:Follicular Fluid / Q000166:cytology; D006107:Granulosa Cells; D006801:Humans; D029589:Seminal Plasma Proteins / Q000276:immunology
chemical_list: D000911:Antibodies, Monoclonal; D002021:Buffers; D002352:Carrier Proteins; C552592:FSIP1 protein, human; D029589:Seminal Plasma Proteins