Co-culture of human extravillous cytotrophoblasts and decidual stromal cells in vitro / 南方医科大学学报

<p><b>OBJECTIVE</b>To develop a convenient method for isolation and purification of human extravillous cytotrophoblasts (EVCTs) and decidual stromal cells (DSCs) and establish a co-culture system.</p><p><b>METHODS</b>The DSCs were digested with trypsin and purified by Percoll gradient. The EVCTs were digested with trypsin and purified by BSA gradient. Immunohischemistry and immunofluorescent study are performed to characterize these isolated cells. The EVCTs and DSCs were placed in Matrigel-coated Transwell upper and lower chamber, respectively, to study the invasive ability of the EVCTs.</p><p><b>RESULTS</b>Immunohischemistry revealed that the purity of EVCTs and DSC exceeded 95%. Cultured EVCTs retained their capacity to invade Matrigel-coated Transwell filters with the invasion index of 3.22-/+0.04.</p><p><b>CONCLUSION</b>This co-culture model established by isolating highly purified EVCTs and DSCs in vitro can be useful for studying the trophoblast invasion mechanisms.</p>

Effect of nitric oxide synthase inhibitor on proteoglycan metabolism in repaired articular cartilage in rabbits / 中华创伤杂志(英文版)

<p><b>OBJECTIVE</b>To study the effect of nitric oxide synthase inhibitor, S-methyl thiocarbamate (SMT), on proteoglycan metabolism in repaired articular cartilage in rabbits.</p><p><b>METHODS</b>Twenty-four male New Zealand white rabbits, aged 8 months and weighing 2.5 kg+/-0.2 kg, were used in this study. Cartilage defects in full thickness were created on the intercondylar articular surface of bilateral femurs of all the rabbits. Then the rabbits were randomly divided into 3 groups (n=8 in each group). The defects in one group were filled with fibrin glue impregnated with recombinant human bone morphogenetic protein-2 (rhBMP-2, BMP group), in one group with fibrin glue impregnated with rhBMP-2 and hypodermic injection with SMT (SMT group) and in the other group with nothing (control group). All the animals were killed at one year postoperatively. The tissue sections were stained with safranine O-fast green and analyzed by Quantiment 500 system to determine the content of glycosaminoglycan through measuring the percentage of safranine O-stained area, the thickness of cartilages and the mean gray scale (average stain intensity). Radiolabelled sodium sulphate (Na(2)(35)SO(4)) was used to assess the proteoglycan synthesis.</p><p><b>RESULTS</b>At one year postoperatively, the percentage of safranine O-stained area, the mean gray scale and the cartilage thickness of the repaired tissues in SMT group were significantly higher than those of BMP group (P<0.01) and the control group (P<0.05). Result of incorporation of Na(2)(35)SO(4) showed that the proteoglycan synthesis in SMT group was higher than those of BMP group and the control group (P<0.01).</p><p><b>CONCLUSIONS</b>SMT, a nitric oxide synthase inhibitor, can significantly increase the content of glycosaminoglycan and proteoglycan synthesis, and computer-based image analysis is a reliable method for evaluating proteoglycan metabolism.</p>