- 期刊
Biological Properties of Tenocytes from Primary Culture: Isolation, Culture and Proliferation
肌腱韌帶細胞之分離、培養與生長特性
並列摘要
Tissue engineering affords the biological substitutes for the compromised tissues, including ligaments, cartilage, bones, etc. There are three major elements in studying tissue engineering including the cells, the scaffold and the bioreactor. The basic property of the cell must be well defined. The cell is the most important component among the three basic components. When the cells are manipulated, the biological activity and characteristics of the cells must be well defined and well known. In the field of ligament tissue engineering, most researchers used the primary-cultured tenocytes to be their cell source. The tenocytes were cultured from anterior circulate ligament, medial collateral ligament, patella tendon, Achilles tendon or flexor tendon of rats, rabbits, chickens and even human. The definition of the biological property of the tenocytes becomes very important because their sources are diverse. There are two major sources of cells that were used in ligaments tissue engineering. One is the cells isolated from primary cell culture, and the other is from the mesenchymal stem cells. In our study, the source of the tenocytes is from the primary cell culture. We took the tendon material from the flexor hallucis tendon of Wistar rats. We used the modified Banes method to isolate and culture the tenocytes successfully. Then we measured the proliferation rate of the tenocytes using two different methods. The first and most direct method is to measure the proliferation of the tenocytes by counting the cell number with the hemocytometer directly. The second method we employed is the incorporated BrdU colorimetry method. We compared the two methods and found their results equivalent. We found that the tenocytes will decrease the proliferation rate gradually from the 13(superscript th)~16(superscript th) generation. Up to the 18(superscript th) generation, the tenocyte proliferation almost stopped. Within 13(superscript th) generation of proliferation, the tenocytes will maintain the phenotype very well. So the tenocyte cultured from the tendon tissue directly is a good material for ligament tissue engineering.