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  • About
  • The Global ETD Search service is a free service for researchers to find electronic theses and dissertations. This service is provided by the Networked Digital Library of Theses and Dissertations.
    Our metadata is collected from universities around the world. If you manage a university/consortium/country archive and want to be added, details can be found on the NDLTD website.
1

Cultivo primario de condrocitos a partir de cartílago hialino articular de Equus Caballus post-mortem

Vegas Albino, Diana Pamela January 2011 (has links)
El objetivo de esta tesis fue obtener un cultivo primario de condrocitos de Equus caballus post-mortem, aislados de cartílago hialino articular esternocostal. Se colectaron un total de 24 de cajas torácicas de caballos beneficiados para consumo humano y animal, obtenidos del Camal de Equinos Casablanca ubicado en el distrito de Pachacamac, Lima; de cada una de ellas se obtuvieron 4 muestras de cartílago de forma semiovalada irregular de 0.8 cm de ancho, 0.6 cm de largo y 0.2 cm de alto aproximadamente. El aislamiento de condrocitos se realizó de forma mecánica, con la técnica de cuchillas cruzadas de Freshney (2005), y de forma enzimática con solución de tripsina 0.05 % con 0.5 g de EDTA, solución de colagenasa 0.2% y 0.04 %. Luego las células fueron cultivados en Basal Medium Eagle (BME) + 10% Suero Bovino Fetal (SBF), BME + 5%SBF, Tissue Culture Medium-199 (TCM-199) + 10%SBF y TCM-199 + 5%SBF; por un periodo de 10 días a 37 °C, 5% de CO 2 y 5% de humedad. Posteriormente, las células fueron resuspendidas de las placas en solución de tripsina (tripsina 0.05%, EDTA 0.2 g.), para contarlas en una cámara de Neubauer y así determinar la tasa de proliferación y la tasa de supervivencia. Se observó un incremento en la proliferación celular tanto en BME como en TCM-199 con 10% de SBF, siendo la tasa de proliferación obtenida para BME+10%SBF de 4,550+/-1,366, para TCM-199+10%SBF de 3,709+/-1,183, para BME+5%SBF de 0,438+/-0,095 y para TCM-199+5%SBF de 0,388+/-0,093; por tanto, el medio de cultivo que obtuvo la mejor tasa de proliferación fue BME + 10%SBF. La tasa de supervivencia no superó el valor de 1 en todos los medios; sin embargo, la que se acercó más a este valor fue la obtenida con BME + 10%SBF (0.987+/-0.025). En conclusión, se logró obtener un cultivo primario de condrocitos a partir de cartílago hialino articular esternocostal de Equus caballus post-mortem, siendo BME + 10% SBF el medio que presentó los mejores resultados. / --- The objective of this thesis was to obtain a primary culture of chondrocytes from postmortem Equus caballus, isolated from hyaline articular sternocostal cartilage. We collected a total of 24 rib cages of benefit horses for human and animal consumption, obtained from Casablanca Equine Camal located in the district of Pachacamac, Lima; from each one we obtained 4 irregular semi-oval samples of 0.8 cm width, 0.6 cm long and 0.2 cm tall. The isolation of chondrocytes was performed mechanically, with the technique of cross blades from Freshney (2005), and so enzymatically with 0.05% trypsin solution with 0.5 g of EDTA, 0.2% and 0.04% collagenase solution. Then they were grown in Basal Medium Eagle (BME) + 10% Fetal Bovine Serum (FBS), BME + 5% FBS, Tissue Culture Medium-199 (TCM-199) + 10% FCS and TCM-199 + 5% FBS; for a period of 10 days at 37 ° C, 5% CO2 and 5% humidity. Subsequently, cells were resuspended from the plates in trypsin solution (trypsin 0.05%, EDTA 0.2 g.), to count them in a Neubauer chamber and to determine the proliferation rate and survival rate, respectively. We observed an increase in cell proliferation in both BME and in TCM-199 with 10% FBS, the proliferation rate obtained for BME +10% FBS was 4.550 + / -1.366, for TCM-199 +10% FBS was 3.709 + / -1.183, for BME +5% FBS was 0.438 + / -0.095 and for TCM-199 + 5 % SBF was 0.388 + / -0.093, therefore the culture medium that obtained the best proliferation rate was BME + 10% FBS. The survival rate did not exceed the value of 1 in all media, but the one closer to this value was the obtained with BME + 10% FBS (0987 + / -0025). In conclusion, it was possible to obtain a primary culture of chondrocytes from hyaline articular sternocostal cartilage of Equus caballus post-mortem, being BME + 10% FBS the medium which had the best results.

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