Practical aspects of donor cell selection for nuclear cloning.
Choosing the right nuclear donor is the most critical decision in cloning by nuclear transfer (NT), or nuclear cloning, because the cloned animal will be a genetic copy of the donor cell genome used for NT. Both donor cell type and cell cycle stage are important methodological parameters and influence nuclear cloning efficiency. Cloning, however, is a multi-step procedure and the exact contribution of the nuclear donor to overall cloning success must be determined in comparative studies. This requires strict standardization of isolation, purification, and culture protocols, and application of stringent identification criteria in order to obtain a homogenous donor cell population. In all these respects, the standards in the cloning field are currently poor. The aim of this review is to provide a brief guideline for the major practical aspects of donor cell selection, cell cycle synchronization and preparation for NT.
Oback
Björn
B
Reproductive Technologies, AgResearch Ltd., Ruakura Research Centre, Hamilton, New Zealand. bjorn.oback@agresearch.co.nz
Wells
David
D
D000818:Animals; D019976:Cloning, Organism; D016678:Genome; D017408:Guidelines as Topic; D053652:Nuclear Transfer Techniques