Quantification of insulin-like growth factor I receptors on granulosa cells with flow cytometry after follicular stimulation | Zendy

Diane De Neubourg | Zendy; A. Robins | Zendy; S. Fishel | Zendy; L. Delbeke | Zendy

Open Access

Quantification of insulin-like growth factor I receptors on granulosa cells with flow cytometry after follicular stimulation

Author(s) -

Diane De Neubourg,

A. Robins,

S. Fishel,

L. Delbeke

Publication year - 1998

Publication title -

human reproduction

Language(s) - English

Resource type - Journals

SCImago Journal Rank - 2.446

H-Index - 226

eISSN - 1460-2350

pISSN - 0268-1161

DOI - 10.1093/humrep/13.1.161

Subject(s) - follicular fluid , flow cytometry , receptor , biology , granulosa cell , insulin like growth factor , follicular phase , monoclonal antibody , ovarian follicle , clone (java method) , endocrinology , medicine , growth factor , andrology , antibody , oocyte , chemistry , microbiology and biotechnology , immunology , biochemistry , embryo , dna

The presence of insulin-like growth factor I (IGF-I) receptors on granulosa cells was investigated by flow cytometric analysis. Granulosa cells were retrieved from follicular fluid after oocyte retrieval during assisted reproduction technology procedures. Whole samples of follicular fluid were pooled and the cellular fraction analysed. In order to analyse granulosa cells only we developed a dual labelling technique whereby granulosa cells were identified as CD45 negative cells, distinguishing them from leukocytes which are CD45 positive. Analysis of the IGF-I receptor was done by staining the sample with a monoclonal anti-IGF-I receptor antibody (alphaIR3 clone) and goat anti-mouse phycoerythrin labelled antibody. After identification of the presence of IGF-I receptors, receptor expression was quantified by calibration of the fluorescence signals. We analysed 10 patients' samples and showed 559-1774 binding sites per granulosa cell with a mean value of 1125 +/- 382 (SD).

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