Application Adipose-devired Stem Cells And Silk Fibroin And Chitosan Scaffold Materials Building Common Cells-stents Complex Experiments

Objective: Research adipose-devired stem cells cells to cartilage cells of cultivating induction method; Silk fibroin and chitosan with as a 3d porous scaffolds materials, research methods and characteristics of its production; Explore the cartilage cells with after induction with silk fibroin and chitosan scaffold materials in vitro reconstruction cells - the feasibility of stent complexMethods:1) Use stick wall separation methods from the human body adipose tissue suction fat stem cells isolated liquid, cultivate amplification, to third generation in BMP and alkaline fibroblast growth factor in cartilage cells of inductive medium induced fat, cartilage cells of stem cells to transformative collagen immunohistochemical detection cells characteristics2)With silk fibroin chitosan for stent raw material, silk after dissolving degum dialysis silk fibroin solution, when mixed with chitosan solution to dissolve, mixed solution freeze-drying legal system into sf/chitosan porous scaffold materials, three-dimensional by volume method for calculation of stents porosity of stents, through the appearance of scanning electron microscopy observation and measurement and performance3)Induction of cells after inoculation of stents in disinfection on material with stents complex, cells joint training scanning electron microscopy (sem) cells - stents complex appearance and internal structure, and the growth of cells on stents inResults:1) Adipose tissue through the digestive get fat cells, and through the medium training three generations of fat decent stem cells in BMP and alkaline fibroblast growth factor induction medium of cartilage, the shape of the cells in the induction training is changed, the characteristics of cartilage cells by long spindle deformation collagen type for polygons, immunohistochemical staining was positive2) Production completed silk fibroin/chitosan 3d porous scaffolds were white, and a cavernous, visible fibrous material surface evenly arrange stents, soft and elastic, pore size uniform, without apparent spike and sag stents greater than 80% porosity with different proportion of stents to make silk fibroin and chitosan with the proportion of the most appropriate 4:6 rod3) Induction of cells after vaccination in culture on the stent joint training, scanning electron microscopy (sem) display cells and stent, the cells can adhesion good stent material inside attached with a growthConclusion:(1) Adipose-devired stem cells in cartilage medium, usually used alone one cell growth factor can be induced in this experiment called cartilage cells of combined application BMP 14 and alkaline fibroblast growth factor two cell growth factor is used alone BMP can better induction fat cartilage cells of stem cells to grow better direction, differentiated cells more densely(2). Silk and chitosan two kind of natural material through a mixture of dissolved get three-dimensional scaffold materials via porous measured pore ratio greater than 80%, helps cells infiltration and nutrient exchange 3d environment also can provide suitable for cells of the microenvironment all sf/chitosan can be as cartilage tissue engineering scaffolds 3d porous scaffolds (3)Adipose-devired stem cells after after induction chondrocytes can very good adhesion in sf and chitosan stents, cells can in surface and internal growth cells in the after induction stent complex。This experiment cell - stents compound can be used as tissue engineering experimental study of cartilage...