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Effective isolation of Sertoli cells from New Zealand rabbit testis
2021
Wen-Qian Zhu | De-Cai Yang | Yu Jiang | Ning-Ning Cai | Rui Yang | Xue-Ming Zhang
Objective: Sertoli cells (SCs) are important sustentacular cells in the seminiferous tubules of the testis. Isolation and identification of SCs are the premise for studying their functions. Since New Zealand rabbit is a stable strain which is widely used for biomedical research and animal farming, this study aimed to develop a simple and effective protocol for SC isolation in New Zealand rabbits. Materials and Methods: The SCs of three 30-day-old New Zealand rabbits were isolated by incu¬bation with enzymatic digestion I (Dulbeccos modified Eagle medium supplemented with 1 mg/ ml collagenase IV and 50 μg/ml DNase I) and digestion II (digestion I + 1 mg/ml hyaluronidase + 1 mg/ml trypsin), as well as differential plating. The cells were enriched and identified by using immunocytochemical staining and reverse transcription polymerase chain reaction analysis. Results: Homogeneous cells were obtained. They presented the typical large cell body and an irregular pyramidal shape after differential plating and passaging. These cells expressed mRNA of the SC marker sex-determining region Y-box 9 (SOX9) instead of the Leydig cell marker StAR. Immunocytochemically, they are positive of SOX9, GATA binding protein 4, and androgen-binding protein. Conclusion: The SCs were enriched from the testicular tissues of prepubertal New Zealand rabbits by a simple and effective protocol, which provides a basis for further theoretical researches and practical applications. [J Adv Vet Anim Res 2021; 8(2.000): 218-223]
Afficher plus [+] Moins [-]Kinematic analysis of cutting horses working a mechanical flag
1989
Clayton, H.M.
High-speed cinematography was used to record the movements of 12 cutting horses performing a standard test with a mechanical flag. Based on their previous competitive performances, horses were classified into 2 groups: group 1, composed of 5 moderately successful or average performers that had won less than $35,000 in purse money; and group 2, composed of 7 highly successful or elite performers that had amassed greater than $35,000 in competition earnings. Analysis of the results indicated that, compared with horses of the average group, the elite horses had faster reaction times in response to the start and cessation of flag movement (P less than 0.01), and were positioned closer to the flag during all stages of the trial (P less than 0.05). Discriminant analysis was used to construct a mathematical formula that could be used to classify an individual horse into 1 of the 2 alternative groups, based on the set of measurements. Two predictor variables were selected that described the maximal distance between the horse and the flag during the run and the part of the body that was moved first in response to the initial flag movement. The accuracy of the predicted group membership, compared with the actual group membership, was 100%.
Afficher plus [+] Moins [-]Separation of monocytes from canine peripheral blood
1989
Kim, J.B. | Lee, B.W. (Chonnam National Univ., Kwangju (Korea R.). Coll. of Veterinary Medicine)
Pure separation of various leukocytes is required for the assessment of their roles in immunological and physiological function. In this study, pure separation of monocytes from canine peripheral blood was attempted. At first, mononuclear cells (PBMC) were separated by ficoll-hypaque gradient method and then monocytes were recovered from PBMC suspensions in sucrose gradient Sol. (PBMC-Sucrose), autologous plasma (PBMC-Plasma) and autologous serum (PBMC-Serum) incubated at 37deg C for 2 hours. In the separation of PBMC by ficoll-hypaque gradient method in canine blood, higher relative centrifugal force (RCF) was required, as high as more than 1,300xg RCF for 40 minutes, for clear formation of PBMC layer than that in human blood as usually used 400xg RCF for 40 minutes. In monocytes-separation from three PBMC suspensions following PBMC separation, recovery-, purity- and viability- rate of monocytes showed better results in PBMC-Plasma and PBMC-Serum than in PBMC-Sucrose suspension, particularly showing better results from PBMC suspensions performed by centrifugation at 1,500xg RCF for 40 minutes
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