The Study Of 3D Co-culture With Fibroblast And Gelatin Sponges

Objective:Primary cultured fibroblasts with tissues in vitro, observing the morphology, growth characteristics and the proliferation of the cells, observeing the co-cultured conditions with absorbable gelatin sponge in vitro. Lay the first stone for the fibroblasts used in tissue engineering in the further study.Methods:4-6 months old New Zealand white rabbits, Primary cultured fibroblasts with Fascia from the hypoderma in vitro Passage to the 3rd generation. Observed the Cell morphology with inverted microscope, Immunohistochemical staining discuss the Laws in cell proliferation with the kits MTT and CCK8 Separated the protein was observed Secreting by the cells, staining withâ… /â…¢collagen. The situations of the Fibroblasts labeled by PKH26, co-cultured with the gelatin sponge In vitro,were observed by the Scanning electronic microscope.Results:Fascia tissue using tissue culture method can be successfully obtained a large number of primary fibroblasts, this method is widely drawn, the process is simple, fibroblasts cultured in vitro proliferation activity. CCK8 and MTT method were used 7 days continuous observation, in the cell cycle proliferation of cells appeared 4-5 days after the plateau (mean 4.1â– 2.0 days and 4.6â– 1.7 days). Comparison of two methods was no significant difference (P>0.05). Immunohistochemical staining were observed under confocal microscope after the cells were typical fibroblast morphology:large cell body, many processes of the spindle or star of flat cells, oval nuclei were regular, multi-nucleoli.â… /â…¢collagen staining. PKH26 fluorescent dye can be a good marker fibroblast cell wall, staining was 100%. Scanning electron microscopy of fibroblasts grown in gelatin sponge support strong, active proliferation.Conclusion:1. Subcutaneous fascia tissue block culture of fibroblasts is a simple way, can get a large number of fibroblasts in vitro 2. Gelatin sponge is a good scaffold ligament tissue engineering, and fibroblast cell sticking rate, good in vitro co-culture.