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Cultures can be maintained by addition of fresh medium or replacement of medium. Alternatively, cultures can be established by centrifugation of the suspension with subsequent resuspension in fresh medium at 1 to 2 x 10⁵ viable cells/mL. Add medium as the cell density increases. Maintain cultures at cell concentrations between 1 x 10⁵ and 1 x 10⁶ viable cells/mL.

Medium Renewal: Add fresh medium every 2 to 3 days. Scrape attached cells if necessary.

Rankin TL, et al. Human ZP3 restores fertility in Zp3 null mice without affecting order-specific sperm binding. Development 125: 2415-2424, 1998. PubMed: 9609824

Greenhouse S, et al. Antibodies to human ZP3 induce reversible contraception in transgenic mice with 'humanized' zonae pellucidae. Hum. Reprod. 14: 593-600, 1999. PubMed: 2118547

Animals were immunized with a human zona pellucida 3 (ZP3) peptide with a C-terminal cysteine (RQPHVMSQWSRSASRNC) coupled to keyhole limpet haemocyanin. The zona pellucida is an extracellular matrix that mediates critical steps in fertilization and early development surrounds all vertebrate eggs, and in mice and humans comprises three glycoproteins: ZP1, ZP2 and ZP3.

Hay, R. J., Caputo, J. L., and Macy, M. L., Eds. (1992), ATCC Quality Control Methods for Cell Lines. 2nd edition, Published by ATCC.

Caputo, J. L., Biosafety procedures in cell culture. J. Tissue Culture Methods 11:223-227, 1988.

Fleming, D.O., Richardson, J. H., Tulis, J.J. and Vesley, D., (1995) Laboratory Safety: Principles and Practice. Second edition, ASM press, Washington, DC.