科研之友
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摘要
The objective of this project is to produce pudu embryos, obtained by means of fibroblast transfer taken from the ear of one animal, to the oocytes of a domestic ruminant such as cattle. In further work, the pudu embryos will be transferred to the uterus of synchronized recipient females of another species. Biopsies 1mm in size were obtained from the outer rim of the ear of two male pudu deer belonging to the Buin Zoological park, Santiago, Chile. The cell lines have been established and kept according to protocols used in cattle. The oocytes are obtained through a puncture in the oocyte-cumulus complex (OCC) from cow ovaries recovered from the butcher. Each oocyte is enucleated and fused with an isolated fibroblast which is inserted beneath the pellucid zone. The fusion of cell membranes is achieved by means of electric shocks. In respect to the timetable, we have observed that a stage of two blastomers is reached on the second day, and morulae of 8 to 16 cells on the third day. On the fourth day it has differenciated as a blastocyst and on the seventh day it finally hacht from the pellucid zone. The obtainment of embryonic blastocytes indicates that it is possible to obtain pudu embryos through heterospecific cloning, even though the percentage of success is relatively low. The viability of the embryos obtained in this manner after in uterus transfer remains still to be verified Los objetivos de este trabajo fueron los de producir embriones de pud迆, obtenidos por la transferencia de n迆cleos de fibroblastos de la oreja de pud迆 en ovocitos de un rumiante dom谷sticos que es el bovino. Para posteriormente en un trabajo futuro proceder a la transferencia de embriones de pud迆, al 迆tero de hembras receptoras sincronizadas de otra especie. Se obtuvieron biopsias de 1 mm aproximadamente del borde externo de la orejas de dos ciervos pudu machos del jard赤n zool車gico Buin-Zoo, Santiago de Chile. Las l赤neas celulares han sido establecidas y conservadas seg迆n los protocolos utilizados para las bovinos. Los ovocitos son obtenidos por punci車n del complejo c迆mulos-ovocito (COC).desde ovarios de vacas recuperados del matadero. Cada ovocito es enucleado y fusionado con un fibroblasto aislado insertado bajo la zona pel迆cida. La fusi車n de membranas celulares es obtenida por choques el谷ctricos. En cuanto a la cronolog赤a, observamos que al segundo d赤a se forma una etapa de dos blast車meras, al tercer d赤a m車rulas de 8 a 16 c谷lulas, y desde el cuarto d赤a se ha diferenciado como blastocisto, el cu芍l al s谷ptimo d赤a termina por eclosionar de la zona pel迆cida La obtenci車n de blasto
