Efficient expression and functional verification of recombinant human type Ⅲ collagen in CHO cells

Collagen is a structural protein and the main component of extracellular matrix. Collagen Ⅲ is twisted from three peptide chains to three spirals, which provides tensile strength and structural integrity for many organs, and participates in regulating basic activities such as cell adhesion, proliferation, migration and differentiation. In this study, PiggyBac transposon was utilized to achieve the efficient expression of recombinant human collagen III in CHO cells. The recombinant collagen was obtained through nickel column purification, and the purified protein was subject to in vitro antioxidant studies, cell proliferation promotion analysis, and structural characterization. ABTS and FRAP experiments showed that it has certain antioxidant activity. CCK8 experiment proved that two kinds of collagen can promote the proliferation of human mesenchymal stem cells. The structure of recombinant collagen was characterized by circular dichroism, and it was confirmed that the expressed collagen could form an ordered triple helix structure, which provided some theoretical basis for its application in food, cosmetics and medical industries.