Details

Autor(en) / Beteiligte

• Li, Weijian
• Liu, Yalan
• Wang, Zhen
• Wang, Li
• Ma, Xiuling
• Wang, Xuguang

Titel

Optimization of handmade cloning technique in sheep and preliminary investigation of umbilical cord mesenchymal stem cells as donor nuclei

Ist Teil von

• Reproduction in domestic animals, 2024-09, Vol.59 (9), p.e14632-n/a

Ort / Verlag

Germany: Blackwell Publishing Ltd

Erscheinungsjahr

2024

Quelle

Wiley Online Library Journals Frontfile Complete

Beschreibungen/Notizen

• Handmade cloning (HMC) has a higher yield and is relatively less difficult to operate compared to traditional micromanipulation cloning. Yet, there are few reports on handmade cloning in sheep. Therefore, this study investigates the key nodes such as AC and DC voltage, denucleation method and fusion method in sheep handmade cloning. In addition, it compares the effects of fibroblasts (FC) and umbilical cord mesenchymal stem cells (UC‐MSCs) of different states as donors on the development of HMC embryos. Furthermore, the effect of different freezing solutions on the survival rate of frozen blastocysts without zona pellucida was also investigated. The results indicate that an AC voltage of 150 V/cm and a DC voltage of 1800 V/cm significantly enhanced the fusion and blastocyst rates (p < .01). The blastocyst rate achieved with umbilical cord MSCs as nucleus donors was significantly higher (40.3%) than that achieved with fibroblasts and differentiated umbilical cord MSCs (21.5%, 22.5%) (p < .01). The highest survival rate was achieved using 20% DMSO + 20% EG for freezing without zona pellucida. In conclusion, the most efficient and pregnant ovine HMC cloning method using 150 V/cm AC, 1800 V/cm DC, knife‐cut denucleation, two‐step fusion and the use of UC‐MSCs as nucleus donors resulted in the highest overall efficiency and pregnancy after transplantation.