RRC ID 43727
著者 Chilvers RA, Bodenburg YH, Denner LA, Urban RJ.
タイトル Development of a novel protocol for isolation and purification of human granulosa cells.
ジャーナル J Assist Reprod Genet
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.
巻・号 29(6)
ページ 547-56
公開日 2012-6-1
DOI 10.1007/s10815-012-9739-5
PMID 22407611
PMC PMC3370046
MeSH 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
IF 2.829
引用数 9
WOS 分野 OBSTETRICS & GYNECOLOGY GENETICS & HEREDITY REPRODUCTIVE BIOLOGY
リソース情報
ヒト・動物細胞 KGN(RCB1154)