276 EXPRESSION OF CD9 AND α6-INTEGRIN IN PORCINE BLASTOCYSTS
M. D. Goissis A , F. R. O. de Barros A , M. G. Marques A , C. M. Mendes A , M. P. Milazzotto A , C. H. C. Viana B , M. E. O. A. Assumpção A and J. A. Visintin AA Laboratory of In vitro Fertilisation, Cloning and Animal Transgenesis, Department of Animal Reproduction, School of Veterinary Medicine and Animal Sciences, University of São Paulo, São Paulo, SP, Brazil;
B Veterinary Medicine Course, PUC Poços de Caldas, Poços de Caldas, MG, Brazil
Reproduction, Fertility and Development 21(1) 235-235 https://doi.org/10.1071/RDv21n1Ab276
Published: 9 December 2008
Abstract
Establishment of embryonic stem cell (ESC) culture in pigs has not been achieved. Verification of pluripotency markers is necessary for validation of a pluripotent cell line. Not all markers observed in ESC from other species are characterized in swine embryos. The objective of this study was to characterize CD9 and α6-integrin expression in porcine blastocysts and to derive porcine ESC using Matrigel. In vitro or in vivo porcine blastocysts were submitted to total RNA extraction for RT-PCR, fixation for immunocytochemistry or immunosurgery for culture of inner cell mass. Expression of Oct-4, CD9, and α6-integrin was detected by PCR. CD9 and α6-integrin PCR products had their nucleotide sequence assessed and compared with public nucleotide database. CD9 product was identical to CD9 porcine sequences and α6-integrin product was similar to human and equine α6-integrin. Immunocytochemistry revealed Oct-4 expression in cytoplasm of the inner cell mass (ICM) and trophoblast cells. CD9 and α6-integrin were observed preferentially on trophoblast cells. No ESC colonies were obtained using co-culture on mouse embryonic fibroblasts (MEF) or on Matrigel. This study describes for the first time expression of CD9 and α6-integrin in porcine blastocysts.
Financial support: Fapesp 05/57314-0.
