Isolation And Determination Of Extracellular Matrix Component Fibronectin And Studies On Its Relationship With Cell Prolifertion And Differentiation

The present study was designed on the basis of establishment of the isolation methods for high yield of purified porcine plasma fibronectin (FN) and high affinity pure FN on rabbit antibody, to investigate the biological effect of FN cell adhesion, proliferation and differentiation, as well as the maturation and release of red blood cells from bone marrow to circulation. In connection with the closed relationship between FN and cytoskeleton, we have also studied its changes before and after cell transformation, cancerization and decancerization, and their possible role in the denucleation process of mammalian late erythroblast.1. Isolation and determination of FN and other matrix components: Electrophoretic purified FN have been isolated from porcine plasma with self made gelatin and heparin-Sepharose 4B affinity columns. The isolated FN showed one band in PAGE, and divided into two bands in SDS-PAGE, which means that the FN consists of two similar subunit chains each with molecular weight of 220 KD and linked together by disulfide bond to form a large molecule of glycoprotein. The purified porcine FN has been shown to be immunologically cross reacted with FNs of other origin, and has found to have strong activity on CHO cell adhesion in serum-free medium. At the same time, we have also isoiated another large glycopro(?)n--Polynectin (PN), which is s(?)milar to FN with respeet to...