Method for dynamically constructing ADSCs (adipose-derived stem cells)-chitosan/ gelatin hydrogel engineered cartilage

Abstract

The invention belongs to the field of biotechnology and tissue engineering and relates to a method for dynamically constructing an ADSCs (adipose-derived stem cells)-chitosan / gelatin hydrogel engineered cartilage. The method comprises the steps of preparing 4% chitosan / gelatin hydrogel serving as a scaffold of the tissue engineered cartilage, and preparing a cartilage induction matrix; making a round stainless steel tripod according to the diameter of a spinner bottle and the height of a stirring rod, and fixing an inert metal wire on the round stainless steel tripod; soaking the scaffold into 75% alcohol for at least three times, then soaking the scaffold into a PBS (phosphate buffer solution), infiltrating into a DMEM (Dulbecco's modified eagle medium), drying in air, sterilizing, transferring into a porous plate, inoculating the scaffold with adipose stem cells of which the number of culture generations is not more than eight at the density of 10<5>-10<7>cells / ml, and culturing in a 5% CO2 incubator with a temperature of 37 DEG C for not less than 2h to obtain an ADSCs-hydrogel complex. Compared with the engineered cartilage obtained by traditional static culture, the engineered cartilage constructed by the method has many advantages and lays a foundation for repairing cartilage defects in animals.

Description

technical field

[0001] The invention belongs to the fields of biotechnology and tissue engineering, in particular to a method for dynamically and stably constructing tissue engineered cartilage. Background technique

[0002] The number of cases of osteochondral defects caused by joint degenerative diseases and trauma is increasing, which brings great pain to many patients. Cartilage has a simple structure without blood vessels, nerves and lymphatic tissue, so its ability to repair itself is limited. Although there are many clinical methods for repairing cartilage defect and trauma, the effect is not ideal, and the function of cartilage tissue cannot be fully restored. The rapid development of cartilage tissue engineering research has brought new hope for the repair of clinical cartilage defects.

[0003] Tissue engineering technology is to inoculate in vitro cultured and expanded cells in a biomaterial that has good biocompatibility and can be degraded and absorbed by the ...