Biopsy of Porcine Blastocysts for Sex Identification and Transcriptome Sequencing Based on Laser-Assisted Microdissection.

Abstract

Pre-implantation embryo biopsy is a well-established technique in large livestock like cattle. However, its application in porcine embryos remains relatively limited. In this study, we successfully obtained trophectoderm cells from porcine IVP blastocysts using laser-assisted microdissection. First, we obtained trophectoderm cells of varying cell numbers (Group 1: less than three cells; Group 2: more than five cells) using laser-assisted microdissection and then evaluated the impact of this dissection procedure on embryo morphology. The morphological observation confirmed the complete normal post-biopsy recovery of blastocysts in the two groups. Furthermore, the expression of proteins associated with cell junction and structural integrity remained unaffected. Comparative analysis of blastocyst cell numbers and apoptotic rates among Group 1, Group 2 and the control group further validated the minimalinvasiveness of the laser-assisted microdissection when cutting less than three cells. The quality and utility of the biopsied trophectoderm cells were confirmed through successful sex identification and transcriptome sequencing. In conclusion, this study successfully established a minimal invasive laser-assisted microdissection technique for porcine blastocysts, enabling pre-implantation sex identification and blastocyst quality assessment.