Abstract
Simple SummarySomatic cell nuclear transfer (SCNT) is a technique used to reproduce individuals from their somatic cell nucleus, which is commonly known as cloning. In SCNT, viable cell lines are usually established from living animals and preserved for future use. In the present study, tissues were collected and cryopreserved (rather than cells) from a suddenly deceased champion show camel. We established fibroblast cell lines from this decade-old vitrified tissue and used them as nuclear donors. Both the in vitro and in vivo matured oocytes were used to produce cloned embryos. Blastocysts were transferred to synchronized recipients to establish pregnancies. A total of 18 pregnancies (5 from in vitro matured oocytes and 13 from in vivo matured oocytes) were established, and 11 live offspring were born (2 from in vitro matured oocytes and 9 from in vivo matured oocytes). We concluded that fibroblast cell lines could be established from long cryopreserved tissues, and these cells could be used as nucleus donors to clone animals.Somatic cell nuclear transfer (SCNT) provides a unique opportunity to reproduce animals with superior genetics. Viable cell lines are usually established from tissues collected by biopsy from living animals in the SCNT program. In the present study, tissues were collected and preserved from a suddenly deceased champion camel. We established cell lines from these decade-old tissues and used them as nuclear donors. After 42 h of in vitro maturation, 68.00 ± 2.40% of oocytes reached the metaphase II (M II) stage while 87.31 ± 2.57% in vivo collected oocytes were matured at collection (p < 0.05). We observed a higher blastocyst formation rate when in vivo matured oocytes (43.45 ± 2.07%) were used compared to in vitro matured oocytes (21.52 ± 1.74%). The live birth rate was 6.45% vs. 16.67% for in vitro and in vivo matured oocytes, respectively. Microsatellite analysis of 13 camel loci revealed that all the SCNT-derived offspring were identical to each other and with their somatic cell donor. The present study succeeded in the resurrection of 11 healthy offspring from the decade-old vitrified tissues of a single somatic cell donor individual using both in vitro and in vivo matured oocytes.
Highlights
The camelids are characterized by tremendously large-scale adaptation capacity and high parameters related to their zootechnical performance in captivity
A total of 348 cumulus oocytes complexes (COCs) were collected from 26 ovaries obtained from the local slaughA total of 348 COCs were collected from 26 ovaries obtained from the local slaughterhouse, and 68.00
We have demonstrated that healthy cloned offspring could be obtained by nuclear transfer using donor nuclei obtained from decade-old vitrified tissues of a suddenly deceased champion show camel named Mabrokan
Summary
The camelids are characterized by tremendously large-scale adaptation capacity and high parameters related to their zootechnical performance in captivity. Achieving the satisfactory efficiency of intra- and inter-species somatic cell cloning both within the taxonomic family Camelidae and in other representatives of ruminant and non-ruminant artiodactyls requires comprehensively recognizing the epigenetic and molecular determinants of successfully generating SCNT-derived embryos, conceptuses, and progeny. The pivotal factors determining the SCNT efficacy encompass epigenetic re-programmability of donor cell nuclear genomes [15,16,17,18,19] and their communication with mitochondrial DNA fractions [20,21,22,23,24] in SCNT-derived oocytes and corresponding embryos
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