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Partial Purification of a Serum Fraction from Fasted Pigs that Inhibits Proliferation of Cultured Myogenic Cells¹

University of Minnesota^,3, St. Paul 55108

Abstract

Sera from pigs fasted as little as 24 h appears to contain a factor(s) that inhibits proliferation of myogenic cells in culture. An inhibitor of myogenic cell proliferation has been partially purified from this sera by using a combination of gel filtration and immunoaffinity chromatography. The inhibitory activity elutes from a Sephacryl S-300 column at a K_av (elution minus void volume divided by total minus void column volume) between .41 and .59. Proteins banding at 76 and 67 kilodaltons appear to predominate on sodium dodecyl sulfate polyacrylamide gels of this fraction. Small quantities of each of these proteins were electrophoretically purified and used to elicit production of anti-76 and anti-67 immunoglobulin G in rabbits. These antibodies were used to prepare anti-76 and anti-67 immunoaffinity columns that were used to further purify the inhibitory activity. The anti-67 column was particularly useful in isolating the inhibitor because it removed mitogens that made detection of the inhibitory activity difficult. The partially purified inhibitor inhibits proliferation of L6 myogenic cells in a concentration-dependent manner. On sodium dodecyl sulfate polyacrylamide gels, the predominant proteins in the inhibitor fraction band at approximately 63 and 61 kilodaltons. Inhibitors of myogenic cell proliferation may play an important role in balancing the effects of positive growth factors.

¹ Published as Paper No. 16911 of the Scientific Journal Series of the Minnesota Agric. Exp. Sta. on research conducted under Minnesota Agric. Exp. Sta. Project No. 4816-83. This work was supported by grants from the USDA (87-CRCR-1-2531), the National Pork Producers Council and Monsanto Company.

² To whom all reprint requests should be sent.

³ Dept. of Anim. Sci.