BACKGROUNDCartilage injury has a very poor capacity for intrinsic regeneration. The cell-based treatment strategy for the cartilage repair using differentiated bone marrow mesenchymal stem cells (BMSCs) is, however, a promising approach to the chondral repair. This study was aimed to explore the chondrogenic potential of the goat BMSCs in the Transwell co-culture system and the poly-laetide-co-glycolide (PLGA) scaffolds.

METHODSThe BMSCs were isolated from the goat iliac crest while the chondrocytes were obtained from the goat's last costal cartilage. In the Transwell co-culture system, the BMSCs co-cultured with chondrocytes were designed as group A, whereas the goat's BMSCs induced with the chondrogenic medium were group B. Both groups A and B were the experimental groups, while group C that only contained BMSCs was the control group. In the PLGA scaffolds co-culture system, BMSCs were seeded into the PLGA scaffolds, which were suspended in the 24-well plate, and the control group was established by presence or absence of chondrocytes at the bottom of the 24-well plate. Toluidine blue staining, Alcian blue staining, collagen II immunofluoresence, collagen II immunochemical staining, collagen I, collagen II, COL2a Q-PCR and osteopontin Q-PCR were used to examine the chondrogenic conditions as well as the expressions of chondrogenic and osteogenic genes.

RESULTSCells isolated from the aspirates of the goat bone marrow proliferated rapidly and gained characteristics of stem cells in Passage 4. However, the differentiations of chondrocytes were not apparent in Passage 3. The results from Toluidine blue staining, collagen II immunofluoresence and PCR showed the transformation of BMSCs to chondrocytes in the Transwell co-culture system and PLGA scaffolds. Although the cartilage gene expressions were upgraded in both chondrogenesis group and co-culture system, the osteopontin gene expression, which represents osteogenic level, was also up-regulated.

CONCLUSIONSThe Transwell co-culture system and the PLGA scaffolds co-culture system can promote the chondrogenic differentiation of the goat's BMSCs, while up-regulated osteopontin gene expression in the Transwell co-culture system implies the osteogenic potential of BMSCs.

Animals ; Bone Marrow Cells ; physiology ; Cell Culture Techniques ; methods ; Chondrocytes ; physiology ; Chondrogenesis ; physiology ; Coculture Techniques ; Goats ; Mesenchymal Stromal Cells ; physiology ; Tissue Engineering ; methods ; Tissue Scaffolds

Objective To investigate the impacts of body mass index (BMI) and age on in vitro fertilization-embryo transfer (IVF) and intracytoplasmic sperm injection (ICSI) treatment in infertile patients without polycystic ovary syndrome (PCOS). Methods A retrospective study of 1426 patients during Jun. 2001 - Nov. 2009 was carried out. Multiple regression was used to analyze the effects of BMI (low weight: BMI＜18.5 kg/m2, normal weight:BMI 18.5-23.99 kg/m2 and over weight-obesity: BMI≥24 kg/m2) and age (young: 20-34 years old, eld: 35-45 years old) on controlled ovarian stimulation (COH)[including:dose and duration of Gn, E2 level on day of human chorionic gonadotropin (HCG) administration, number of oocytes collected and full-grown follicles ],number of fertilization, cleavage, two-pronucleus, normal embryos and cryopreserved embryos and clinical pregnancy outcome. Results ( 1 ) Gn dose for the patients whose age were 35 and the above,had a positive correlation with age (P＜0.001), 12.70% of the total variation of Gn dose was related to age (standardized partial regression coefficient was 0.343). (2) Estradiol level on day of HCG administration had a negative correlation with BMI in overweight-obesity patients, and so were the patients whose age were 35 and above (P value respectively lower than 0.037 and 0.018). 0.80% of the total variation of estradiol (HCG day) is related to age and overweight-obesity while age took greater proportion (standardized partial regression coefficients were 0.066 and 0.058 respectively). (3)For older patients, age appeared to have negative relationships with duration of Gn and number of oocytes collected, full-grown follicles, fertilization, cleavage, two-pronucleus, normal embryos and cryopreserved embryos (P＜0.05). (4)Compared to young-normal weight patients, the odds ratio of pregnancy in eld-low weight and eld-overweight-obesity patients were 0.482 and 0.529 (P＜0.05)respectively. Conclusion Age, but not the BMI, had significant effects on IVF/ICSI treatment. It seems that factors as losing weight before IVF or ICSI treatment effective in reducing the dose of Gn.