Abstract
Studies were conducted to explore the possibility of employing dromedary camel (Camelus dromedarius) oocytes as recipient cytoplasts for the development of interspecies somatic cell nuclear transfer (iSCNT) embryos using skin fibroblast cells of an adult Bactrian camel (Camelus bactrianus) and Llama (Llama glama) as donor nuclei. Also, the embryos reconstructed with Bactrian cells were transferred into the uterus of synchronized dromedary camel recipients to explore the possibility of using them as surrogate mothers. Serum-starved skin fibroblast cells were injected into the perivitelline space of enucleated mature oocytes, collected from super-stimulated dromedary camels, and fused using an Eppendorf electroporator. After activation with 5μM ionomycin and 6-dimethylaminopurine, they were cultured at 38.5°C in an atmosphere of 5% CO2, 5% O2, and 90% N2 in air. In experiment 1, Day 7 blastocysts were stained with Hoechst to count their cell numbers, while in experiment 2, they were transferred to synchronized dromedary recipients. A lower number (P < 0.05) of blastocysts were obtained from reconstructs utilizing fibroblast cells from Llama when compared with those reconstructed with dromedary and Bactrian fibroblast cells. However, no difference was observed in their cell numbers. In experiment 2, a higher (P < 0.05) proportion of blastocysts were obtained from the cleaved embryos reconstructed with Bactrian fibroblast cells when compared to those reconstructed with dromedary cells. Twenty-six Day 7 blastocysts reconstructed with Bactrian cells were transferred to 23 synchronized dromedary recipients with 5 pregnancies established on Day 30, however, only one of the pregnancies developed to term and a healthy calf weighing 33 kgs was born after completing 392 days of gestation. Unfortunately, the calf died on day 7 due to acute septicemia. In conclusion, the present study reports, for the first time, birth of a cloned Bactrian calf by iSCNT using dromedary camel as a source for oocytes as well as a surrogate for carrying the pregnancy to term.
Highlights
Our planet has suffered from a progressive reduction in biodiversity during the last century with 5,485 animal species threatened with extinction, including 180 species of mammals according to IUCN—World Conservation Union [1]
In experiment 1, a total of 42 and 58 embryos were reconstructed with llama (LM-SKF) and Bactrian (BT-SKF) fibroblast cells, while as 57 embryos were reconstructed with dromedary (SKF-1) fibroblast cells
The present study reports, for the first time, birth of a cloned Bactrian calf by interspecies somatic cell nuclear transfer (iSCNT) using dromedary camel as a source for oocytes as well as a surrogate for carrying the pregnancy to term
Summary
Our planet has suffered from a progressive reduction in biodiversity during the last century with 5,485 animal species threatened with extinction, including 180 species of mammals according to IUCN—World Conservation Union [1]. The report of the first mammal produced by somatic cell nuclear transfer (SCNT) [2] raised the possibility that this biotechnology technique could be used to save and preserve critically endangered animal species and even could help to restore extinct species [3,4]. It seems to be more appropriate to help in conserving living species that are close to being extinct than those already extinct. These possibilities led to the establishment of DNA banks [5] from endangered species for their possible use in SCNT studies. The only option is to use a closely related domestic animal species as oocyte donors as well as surrogate mothers to carry the cloned embryos to term. The ooplasm of domestic cats supports development of African wild cat [7], domestic cattle oocyte cytoplasm supported proliferation of a gaur [8] and canine oocyte cytoplasm supports development of endangered wolves [9] and a mouflon was produced by iSCNT using a goat as a surrogate mother to carry the fetus to term [10]
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