Partial purification of smooth muscle cell derived growth factor

The serum free medium conditioned by cultured rabbit aortic smooth muscle cells was partially purified using ultrafiltration and heparin affinity chromatography. Incorporation of [3H]-thymidine (3H-TdR) into cell DNA was used to measure the mitogenic activity of the fractions from chromatography for NIH 3T3 fibroblasts. The molecular weight and the iso-electric point of these fractions were determined by NaDodSO4-polyacrylamide gel electrophoresis (SDS-PAGE) and iso-electric focusing, respectively. The results showed that the protein eluted in 1. 0–1. 6 mol/L NaCl from the heparin-Sepharose was mitogenic for 3T3 cells, and this protein had a molecular weight of 22. 8–26. 7 ku and an iso-electric point of about 4. 6. The fact that the above-mentioned biochemical properties differed from that of PDGF, IGF and FGF suggests that this mitogenic protein may be a separate growth factor.