Abstract:

AIM： To confirm the call source of cartilage defect after bone marrow mesenchymal stem calls （MSCs） transplantation, so as to know the recovery process. METHODS： The experiment was conducted in the Institute of Bioengineering, Zhejiang Academy of Medical Sciences from January 2006 to January 2007. ①Three-month-old New Zealand Rabbit of clean grade and about 2 kg was selected （experimental animal center of Zhejiang Province）. Two pairs of PCR primers were designed by using the sex-determining gene sequence. MSCs were isolated from the male New Zealand Rabbit and cultured. The cartilage of knee joint of female rabbits was exposed to establish the full-thickness cartilage defect of 4.5 mm in diameter and 3.5 mm in depth by electrodrill. One side was implanted with MSCs/collagen sponge, and the other with collagen sponge only. At postoperatively 1, 2, 3, 4, and 5 months, the total DNA was extracted from the cartilage defects for PCR amplification to identify whether there was sex-determining gene in the implants, so as to confirm the call source. RESULTS： At 1, 2, 3, 4, and 5 months after transplantation, there was a band （about 200 bp） of DNA extracted from the tissues implanted with MSCs and collagen sponge by PCR; it was of the same size as the positive control, but no band was found in the other side. The sequencing of PCR amplified positive sample showed that the base sequence of every sample was uniform, and accorded with the standard sequence of Y-genome gb ｜AY785433.1｜ from GenBank. The sequencing homology was 100%. CONCLUSION： Male tissues are found in the defects implanted with female MSCs and collagen sponge, indicating that there are chondrocytes that are differentiated from the implanted male MSCs in new formation cartilage.[著者文摘]