Experimental method for establishing tissue engineering cartilage in vitro by taking knee-joint cartilage unit of rabbit as seed cell

Abstract

The invention belongs to the technical field of in-vitro establishment of tissue engineering cartilages, more particularly to an experimental method for establishing tissue engineering cartilage in vitro by taking a knee-joint cartilage unit of a rabbit as a seed cell. The experimental method comprises the following steps of: taking a keen joint of a rabbit, repeatedly cutting the joint cartilage into fragmentized tissues, washing, removing supernatant away and putting in a sterile conical flask for later use; adding dispase and II-type collagenase into the conical flask according to 12mL DMEM-F12 culture solution / g cartilage, stirring and digesting to form a digested suspension solution; filtering and centrifuging the digested suspension solution, adding a primary chondrocyte culture solution to prepare a sterile chondrocyte suspension solution, and carrying out sodium alginate gel three-dimensional culture on the obtained cartilage unit to construct the tissue engineering cartilage. The invention has the beneficial effects that: the cartilage unit has stable form and multiplication situation in the in-vitro long-term culturing process, the physiological function of substrate ingredients, such as secreted II-type collagen, and the like is improved, and the effect of repairing cartilage injury by adopting a tissue engineering method is expectedly accelerated.

Description

technical field [0001] The invention belongs to the technical field of in vitro construction of tissue engineered cartilage, and specifically relates to an experimental method for constructing tissue engineered cartilage in vitro by using rabbit knee articular cartilage units as seed cells. Background technique [0002] During the in vitro construction of tissue-engineered cartilage, the selection of ideal seed cells is the most critical and difficult point in current research. At present, in domestic and foreign literature reports, the most commonly used seed cells for articular cartilage tissue engineering are chondrocytes. The conventional enzymatic digestion method in vitro is: 0.4% Pronase enzyme is used to digest 12mL DMEM-F12 culture solution / g cartilage at 37°C for 90 minutes. After replacing the culture solution, 0.025% type II collagenase is digested overnight to obtain knee joint chondrocytes. However, in the process of in vitro culture and subculture of chondroc...

AI Technical Summary

Problems solved by technology

However, in the process of in vitro culture and subculture of chondrocytes, the viscoelasticity gradually decreases, the fragility increases, and the function of chondrocytes degenerates relatively quickly. During the process of repairing cartilage defects in vivo, hyaline cartilage tissue that conforms to the characteristics of normal cartilage cannot be formed, which has restricted the development of tissue engineering technology. one of the biggest obstacles