A method for prolonged survival of primary cell lines
- Michel A. Horisberger
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We have established a means for prolonged survival of primary cell cultures and establishment of continuous cell lines without genetic manipulations. Primary cultures of granulosa cells degenerate rapidly in vitro by a spontaneous onset of apoptotic cell death. Earlier attempts to circumvent this limitation have included transformation with oncogenes, spontaneous immortalization of primary cultures, and chemical carcinogenesis. We have found that addition of a complex of growth-promoting compounds, carrier proteins, and factors isolated from porcine follicular fluid to standard culture medium allows, reproducibly, the establishment of continuous porcine primary granulosa cell lines with genetic stability. This same supplement allows the prolonged survival of primary cell cultures derived from adult rat ovaries. The rat ovary primary cultures consisted of mixed phenotypes, including epithelial, neuron-like, and mesenchymal cell types. Numerous cells stain positive for alkaline phosphatase in these cultures. Other primary cell lines were established from embryonic rat liver and from adult rat lungs, using the same supplement. The survival effect is reversible because cells degenerate when the supplement is removed. Therefore, the cell lines have neither acquired properties of a tumor cell line nor have they been immortalized by a virus infection. We expect that our approach will open the door to prolonged survival of other primary cell types.
- Andersen, M. M.; Krøll, J.; Byskov, A. G.; Faber, M. Protein composition in the fluid of individual bovine follicles. J. Reprod. Fertil. 48:109–118; 1976. CrossRef
- Bukovsky, A.; Svetlikova, M.; Caudle, M. R. Oogenesis in cultures derived from adult human ovaries. Reprod. Biol. Endocrinol. 3:17–29; 2005. CrossRef
- Chang, S. C. S.; Jones, J. D.; Ellefson, R. D.; Ryan, R. J. The porcine ovarian follicle: I. Selected chemical analysis of follicular fluid at different developmental stages. Biol. Reprod. 15:321–328; 1976. CrossRef
- Chedrese, P. J.; Rodway, M. R.; Swan, C. L.; Gillio-Meina, C. Establishment of a stable steroidogenic porcine granulosa cell line. J. Mol. Endocrinol. 20:287–292; 1988. CrossRef
- Donovan, P. J.; Gearhart, J. The end and the beginning for pluripotent stem cells. Nature 414:92–97; 2001. CrossRef
- Horisberger, M. A. Establishment of reversibly immortalized primary granulosa cell lines. Proceedings of the 37th Annual Meeting of the Union of the Swiss Societies for Experimental Research [abstract P019]; 2005.
- Hu, C.-L.; Cowan, R. G.; Harman, R. M.; Porter, D. A.; Quirk, S. M. Apoptosis of bovine granulosa cells after serum withdrawal is mediated by FAS antigen (CD95) and FAS ligand. Biol. Reprod. 64:518–526; 2001. CrossRef
- Joseph, N. M.; Morrison, S. J. Toward an understanding of the physiological function of mammalian stem cells. Develop. Cell 9:173–183; 2005. CrossRef
- Larsen, W. J.; Wert, S. E.; Chen, L.; Russell, P.; Hendrix, E. M. Expansion of the cumulus-oocyte complex during the preovulatory period: possible roles in oocyte maturation, ovulation and fertilization. In: Familiari, G.; Makabe, S.; Motta P. M., ed. Ultrastructure of the ovary. Boston: Kluwer Academic; 1991:45–61.
- Loo, D. T.; Fuquay, J. I.; Rawson, C. L.; Barnes, D. W. Extended culture of mouse embryo cells without senescence: inhibition by serum. Science 236:200–202; 1987. CrossRef
- Matsui, Y.; Toksoz, D.; Nishikawa, S.; Nishikawa, S.; Williams, D.; Zsebo, K.; Hogan, B. L. M. Effect of Steel factor and leukemia inhibitory factor on murine primordial germ cells in culture. Nature 353:750–752; 1991. CrossRef
- Matzuk, M. M.; Burns, K. H.; Viveiros, M. M.; Eppig, J. J. Intercellular communication in the mammalian ovary: oocytes carry the conversation. Science 296:2178–2180; 2002. CrossRef
- Motta, P. M.; Takeva, Z.; Nesci, E. Ultrastructural and histochemical study of junctions between follicular cells and the oocyte during development of ovarian follicle [in French]. Acta Anat. 80:537–562; 1971. CrossRef
- Motta, P. M.; Nottola, S. A.; Familiari, G.; Makabe, S.; Stallone, T.; Macchiarelli, G. Morphodynamics of the follicular-luteal complex during early ovarian development and reproductive life. Int. Rev. Cytology 223:178–288; 2003.
- Oliver, R. H.; Chen, G. D.; Yeh, J. Follicular atresia. In: Knobil, E.; Neil, J. D., ed. Encyclopedia of reproduction. Vol. 2. London: Academic Press; 1999:373–375.
- Payer, A. F. Permeability of ovarian follicles and capillaries in mice. Am. J. Anat. 142:295–318; 1975. CrossRef
- Polejaeva, I. A.; Chen, S. H.; Vaught, T. D., et al. Cloned pigs produced by nuclear transfer from adult somatic cells. Nature 407:86–90; 2000. CrossRef
- Rodgers, H. F.; Lavranos, T. C.; Vella, C. A.; Rodgers, R. J. Basal lamina and other extracellular matrix produced by bovine granulosa cells in anchorage-independent culture. Cell Tissue Res. 282:463–471; 1995.
- Rubin, H. The disparity between human cell senescence in vitro and lifelong replication in vivo. Nature Biotechnol. 20:675–681; 2002. CrossRef
- Shim, H.; Gutiérrez-Adan, A.; Chen, L-R.; BonDurant, R. H.; Behhoodi, E.; Anderson, G. B. Isolation of pluripotent stem cells from cultured porcine primordial germ cells. Biol. Reprod. 57:1089–1095; 1997. CrossRef
- Stojkovic, M.; Lako, M.; Strachan, T.; Murdoch, A. Derivation, growth and applications of human embryonic stem cells. Reproduction 128:259–267; 2004. CrossRef
- Tilly, J. L.; Billig, H.; Kowalski, K. I.; Hsueh, A. J. Epidermal growth factor and basic fibroblast growth factor suppress the spontaneous onset of apoptosis in cultured rat ovarian granulosa cells and follicles by a tyrosine kinase-dependent mechanism. Mol. Endocrinol. 6:1942–1950; 1992. CrossRef
- Yanagishita, M.; Rodbard, D.; Hascall, V. C. Isolation and characterization of proteoglycans from porcine ovarian follicular fluid. J. Biol. Chem. 254:911–920; 1979.
- A method for prolonged survival of primary cell lines
In Vitro Cellular & Developmental Biology - Animal
Volume 42, Issue 5-6 , pp 143-148
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- primary cell lines
- follicular fluid
- granulosa cells
- cell proliferation
- Industry Sectors
- Author Affiliations
- 1. Aivogen AG, Engelgasse 77, CH-4052, Basel, Switzerland