Live birth of a Pashmina goat kid after transfer of handmade cloned embryos.

Abstract

The present study aimed to establish a zona free (handmade cloning) embryo production system for Pashmina goat embryos. Abattoir derived oocytes were matured in vitro; after maturation, oocytes were enucleated and fused with somatic cells derived from an adult Pashmina goat tissue. The reconstructs were activated using a calcium ionophore-DMAP procedure. The embryos were distributed into two experimental groups. In Experiment 1, the embryos were cultured in one of the following four culture media (i) G1.G2 media (ii) Modified synthetic oviduct fluid (mSOF) (iii) Research vitro cleave media (RVCL) and (iv) Embryo development media (EDM), and were cultured for 7 days. The cleavage rates in G1.G2, RVCL, and mSOF were higher than those in EDM (86.8, 82.4, 77.3, and 68.8%, respectively). Blastocyst rates were higher in RVCL than those in mSOF, EDM, and G1.G2 (15.0, 10.5, 4.9, and 2.2%, respectively). In experiment 2, the embryos were cultured in five different culture systems: (i) Flat surface (FS), (ii) Well in drop (WID), (iii) Well of well (WOW), (iv) Micro drop, and (v) Hanging drop, for 7 days. The cleavage rates in FS and WID were higher than those in WOW, Micro drop, and Hanging drop (84.3, 81.2, 73.6, 73.5, and 70.3%, respectively). The blastocyst rates were higher in WID than those in WOW, Micro drop, Hanging drop, and FS systems (21.6, 13.7, 11.5, 10.9, and 3.9%, respectively). The embryos produced in experiment 2 were transferred to synchronized recipients. Of the three pregnancies established on day 40, one resulted in the delivery of a healthy Pashmina kid.