Optimization of donor cell cycle synchrony, maturation media and embryo culture system for somatic cell nuclear transfer in the critically endangered Vietnamese Ỉ pig

• Published in: Theriogenology Vol. 166; pp. 21 - 28
• Main Authors: Nguyen, Van Khanh, Somfai, Tamas, Salamone, Daniel, Thu Huong, Vu Thi, Le Thi Nguyen, Huong, Huu, Quan Xuan, Hoang, Au Thi, Phan, Hieu Trung, Thi Pham, Yen Kim, Pham, Lan Doan
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 01.05.2021
• Subjects: Culture system; Embryo production; Embryo transfer; Somatic cell nuclear transfer; Vietnamese Ỉ pig; Embryo transfer; Culture system; Somatic cell nuclear transfer; Embryo production; Vietnamese Ỉ pig
• ISSN: 0093-691X; 1879-3231
• DOI: 10.1016/j.theriogenology.2021.02.008

Our aim was to establish an efficient culture system to produce embryos by SCNT of the endangered Vietnamese Ỉ pig. Reducing the serum concentration from 10.0% to 0.2% during culture efficiently synchronized Ỉ pig fibroblasts used as donor cells at the G0/G1 stage. Oocyte maturation in a defined porcine oocyte medium (POM) supplemented with EGF and gonadotrophins resulted in higher cleavage and blastocyst rates compared with a non-defined POM containing pig follicular fluid (but without EGF) and both the defined and non-defined variants of NCSU-37. For embryo culture PZM3 and PZM5 media were superior to NCSU-37, in terms of the percentage of cleaved embryos. Addition of serum to PZM3 medium on Day 5 of culture (Day 0 = SCNT) improved blastocyst development. When SCNT embryos were transferred at the blastocyst stage, 7 of 11 recipients became pregnant. However, live offspring were not obtained. In conclusion, we established a system for the production of Ỉ pig embryos by SCNT and achieved blastocyst production rate at 26.4% by improving culture systems for donor cells, oocytes and embryos culture. Transfer of embryos resulted in pregnancies; however, live offspring were not obtained. •Adult Ỉ pig fibroblasts were successfully and reversibly arrested at G0/G1 phase of the cell cycle by serum deprivation.•EGF was superior to pFF for the supplementation of POM during IVM of Ỉ pig oocytes for SCNT.•The highest Ỉ pig blastocyst development rate was achieved in PZM3 medium supplemented with FBS.•The transfer of SCNT embryos resulted in pregnancies; however, pregnant recipients died during an ASF infection outbreak.