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Characterization of immortalized mouse granulosa cell lines

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Summary

Cell cultures of primary mouse granulosa cells were transfected with a v-myc-containing plasmid, and the resulting stable cell lines were tested for their steroidogenic properties and physiologic status. Granulosa cells were obtained from 22-day-old NMRI mice injected with 8 IU pregnant mare serum gonadotropin i.p. 2 days earlier. In Passage 1 the cells were transfected with pSVv-myc using calcium phosphate precipitation or lipofectin. The 3β- and 17β-hydroxy steroid dehydrogenase activity was visualized in control cultures. The three cell lines obtained have been in culture for over 1 yr and have been subcultured for more than 90 passages. The cell line GRM01, with a doubling time of 37±3 h and a diploid modal chromosome number, produced progesterone, estradiol, as well as inhibinlike and activinlike material under basal conditions. A combination of follicle-stimulating hormone and luteinizing hormone was able to increase the secretion of progesterone. GRM01L, a fast growing clone of the GRM01 line with a doubling time of 10±1 h, retained only the capacity to produce activinlike material and transforming growth factor-β, and it was the only one with a tumorigenic capacity. Epidermal growth factor, insulin, and interleukin-6 were able to induce the [3H]thymidine incorporation into DNA in these two cell lines. GRM02, with a doubling time of 36±2 h and a hypertriploid modal chromosome number, produced progesterone and activinlike and inhibinlike material. Follicle-stimulating hormone and luteinizing hormone were able to enhance the secretion of progesterone. For this cell line, only insulin was shown to induce [3H]thymidine incorporation into DNA.

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  1. Innogenetics N.V., Industriepark Zwijnaarde 7, Box 4, B-9052, Ghent, Belgium

    Tony W. Briers, Andre Van De Voorde & Hugo Vanderstichele

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  1. Tony W. Briers
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  2. Andre Van De Voorde
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  3. Hugo Vanderstichele
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Briers, T.W., Van De Voorde, A. & Vanderstichele, H. Characterization of immortalized mouse granulosa cell lines. In Vitro Cell Dev Biol - Animal 29, 847–854 (1993). https://doi.org/10.1007/BF02631362

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