The Role Of The Regenerated Antheraea Penyi Silk Fibroin In The Proliferation And Differentiation Of Mouse Mesenchymal Stem Cells In Vitro

Silk fibroin is a kind of natural macromolecules, and is used as the surgical sutures with more than 100-year history. Recently, Silk fibroin is applied as a new type of biomaterial more and more broadly because of its unique characteristics including low immunogenicity, good histocompatibility, and slow rate of biodegradation and so on. Except for silk fibroin of domestic silkworm, silk fibroin of antheraea penyi is an important member of silk fibroin. Similarly to silk fibroin of domestic silkworm, silk fibroin of antheraea penyi is mainly composed of glycine and alanine. But silk fibroin of antheraea penyi is better than silk fibroin of domestic silkworm in supporting cell adherence and growth because it contains more alanine and is rich in arginine- glycine- asparticacid (RGD sequence). Mesenchymal stem cells ( MSCs) are defined as pluripotent cells and have the ability to differentiate into multiple mesodermal cells. These cells are uncommitted precursor cells with homing ability, and have extensive renewal potential. MSCs are the idea seeds for tissue engineering because these cells are easily to obtain and enrich, and are not involved in ethical problems. Researchers are focused on the interaction of seed cells and matrix material in tissue engineering and its mechanism. The adherent ability is one of the critical characteristics of a biomaterial matrix because the applicable biomaterial should have an idea ability to adhere to seed cells. In this study, to test the supporting ability of regenerated silk fibroin of antheraea penyi (A penyi SF) to stem cells, we studied the growth and differentiation of murine MSCs cell line C3H10T1/2 and amniotic fluid stem cells (mAFS) that cultured onA penyi SF in vitro. Partâ… The Role of the Regenerated Antheraea Penyi Silk Fibroin in the Proliferation and Differentiation of Mouse Mesenchymal Stem Cell-line in VitroObjective In order to provide the experimental evidence for applying the regenerated antheraea penyi silk fibroin to tissue engineering, we observe the proliferation and differentiation of the mouse mesenchymal stem cell line(C3H10T1/2) on the regenerated antheraea penyi silk fibroin membrane in vitro.Methods Four biomaterials, including the regenerated silk fibroin of domestic silkworm (B mori SF), the regenerated silk fibroin of antheraea penyi (A penyi SF), collagen and the cell culture plastic, were used in the study. The ES cell line C3H10T1/2 cells that were cultured on the four biomaterials, and their adhesion ability, proliferation, differentiation and the phenotypes of were investigated by the observing under inverted microscope, immunostaining and FACS, and MTT assay.Result There is no difference of morphology and phenotype of C3H10T1/2 cells after cultured on these four different materials. The MTT assay showed that C3H10T1/2 cells proliferated more rapidly on the antheraea penyi material than that on the other 3 biomaterials after sixth day's culture(P<0.05). Furthermore, C3H10T1/2 cells can be successfully induced and differentiated into adipocytes. Conclusion These results show that the regenerated silk fibroin of antheraea penyi provide an suitable support for the growth and differentiation of C3H10T1/2 cell, and exhibit good cellular compatibilities.Part II The Role of the Regenerated Antheraea Penyi Silk Fibroin in the Growth of Mouse Amniotic Fluid Stem Cells in VitroObjective To provide the experimental evidence for applying the regenerated antheraea penyi silk fibroin to tissue engineering, we discussed the growth of the mouse amniotic fluid stem cells from GFP mice(GFP-mAFS) on the regenerated antheraea penyi silk fibroin membrane in vitro. Methods The regenerated silk fibroin of domestic silkworm (B mori SF) and the regenerated silk fibroin of antheraea penyi (A penyi SF) were used in the study. The GFP-mAFS cells were cultured on both biomaterials, and their adhesion ability and growth were investigated by the observing under inverted microscope.Result There is no influence of difference of regenerated silk fibroin of antheraea penyi on the morphology and adherent ability of GFP-mAFS cells.Conclusion These results show that the regenerated silk fibroin of antheraea penyi provide an good support for the growth and adhrence of mAFS, and exhibit good cellular compatibilities.