Genomic Stability of Lyophilized Sheep Somatic Cells before and after Nuclear Transfer

PLoS One

Domenico Iuso¹  Marta Czernik¹  Federica Zacchini¹  Fiorella Di Egidio¹  Silvestre Sampino¹  Grazyna Ptak¹  Pasqualino Loi¹  Jacek A. Modlinski²  Zdzislaw Smorag³  Michal Bochenek³ 

[1] Department of Comparative Biomedical Sciences, University of Teramo, Teramo, Italy;Department of Experimental Embryology, Institute of Genetics and Animal Breeding, Polish Academy of Sciences, Jastrzebiec, Poland;National Research Institute of Animal Production, Balice, Poland

关键词: Embryos;    Lymphocytes;    Oocytes;    DNA damage;    DNA repair;    Blastocysts;    DNA fragmentation;    Cell enucleation;   

DOI  :  10.1371/journal.pone.0051317

学科分类：医学（综合）

来源: Public Library of Science

【 摘 要 】

The unprecedented decline of biodiversity worldwide is urging scientists to collect and store biological material from seriously threatened animals, including large mammals. Lyophilization is being explored as a low-cost system for storage in bio-banks of cells that might be used to expand or restore endangered or extinct species through the procedure of Somatic Cell Nuclear Transfer (SCNT). Here we report that the genome is intact in about 60% of lyophylized sheep lymphocytes, whereas DNA damage occurs randomly in the remaining 40%. Remarkably, lyophilized nuclei injected into enucleated oocytes are repaired by a robust DNA repairing activity of the oocytes, and show normal developmental competence. Cloned embryos derived from lyophylized cells exhibited chromosome and cellular composition comparable to those of embryos derived from fresh donor cells. These findings support the feasibility of lyophylization as a storage procedure of mammalian cells to be used for SCNT.

【 授权许可】

CC BY