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Optimization of handmade cloning technique in sheep and preliminary investigation of umbilical cord mesenchymal stem cells as donor nuclei.
Li, Weijian; Liu, Yalan; Wang, Zhen; Wang, Li; Ma, Xiuling; Wang, Xuguang.
Afiliación
  • Li W; College of Animal Science, Xinjiang Agricultural University, Urumqi, China.
  • Liu Y; College of Animal Science, Xinjiang Agricultural University, Urumqi, China.
  • Wang Z; College of Animal Science, Xinjiang Agricultural University, Urumqi, China.
  • Wang L; College of Animal Science, Xinjiang Agricultural University, Urumqi, China.
  • Ma X; College of Animal Science, Xinjiang Agricultural University, Urumqi, China.
  • Wang X; College of Animal Science, Xinjiang Agricultural University, Urumqi, China.
Reprod Domest Anim ; 59(9): e14632, 2024 Sep.
Article en En | MEDLINE | ID: mdl-39279335
ABSTRACT
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.
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Cordón Umbilical / Blastocisto / Clonación de Organismos / Técnicas de Transferencia Nuclear / Células Madre Mesenquimatosas / Fibroblastos Límite: Animals / Pregnancy Idioma: En Revista: Reprod Domest Anim Asunto de la revista: MEDICINA REPRODUTIVA / MEDICINA VETERINARIA Año: 2024 Tipo del documento: Article País de afiliación: China Pais de publicación: Alemania
Texto completo
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Texto completo: 1 Colección: 01-internacional Base de datos: MEDLINE Asunto principal: Cordón Umbilical / Blastocisto / Clonación de Organismos / Técnicas de Transferencia Nuclear / Células Madre Mesenquimatosas / Fibroblastos Límite: Animals / Pregnancy Idioma: En Revista: Reprod Domest Anim Asunto de la revista: MEDICINA REPRODUTIVA / MEDICINA VETERINARIA Año: 2024 Tipo del documento: Article País de afiliación: China Pais de publicación: Alemania