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Vertebrate reproductive science and technology
RESEARCH ARTICLE

Hand-made cloning approach: potentials and limitations

G. Vajta A C , P. M. Kragh A B , N. R. Mtango A and H. Callesen A
+ Author Affiliations
- Author Affiliations

A Section of Reproductive Biology, Department of Animal Breeding and Genetics, Danish Institute of Agricultural Sciences, DK-8830 Tjele, Denmark.

B Department of Human Genetics, University of Aarhus, 8000 Aarhus C, Denmark.

C Corresponding author. Email: gabor.vajta@agrsci.dk

Reproduction, Fertility and Development 17(2) 97-112 https://doi.org/10.1071/RD04116
Submitted: 1 August 2004  Accepted: 1 October 2004   Published: 1 January 2005

Abstract

Two major drawbacks hamper the advancement of somatic cell nuclear transfer in domestic animals. The first is a biological problem that has been studied extensively by many scientists and from many viewpoints, including the cell, molecular and developmental biology, morphology, biochemistry and tissue culture. The second is a technical problem that may be responsible for 50% or more of quantitative and/or qualitative failures of routine cloning experiments and is partially the result of the demanding and complicated procedure. However, even the relatively rare documented efforts focusing on technique are usually restricted to details and accept the principles of the micromanipulator-based approach, with its inherent limitations. Over the past decade, a small alternative group of procedures, called hand-made cloning (HMC), has emerged that has the common feature of removal of the zona pellucida prior to enucleation and fusion, resulting in a limited (or no) requirement for micromanipulators. The benefits of HMC are low equipment costs, a simple and rapid procedure and an in vitro efficiency comparable with or higher than that of traditional nuclear transfer. Embryos created by the zona-free techniques can be cryopreserved and, although data are still sparse, are capable of establishing pregnancies and resulting in the birth of calves. Hand-made cloning may also open the way to partial or full automation of somatic cell nuclear transfer. Consequently, the zona- and micromanipulator-free approach may become a useful alternative to traditional cloning, either in special situations or generally for the standardisation and widespread application of somatic cell nuclear transfer.

Extra keywords: cattle, nuclear transfer, pig, sheep, zona free.


Acknowledgments

We are very grateful for all friends and colleagues who participated in the development of the HMC method. Among many others, we would thank Teija T. Peura, Ian M. Lewis, Tayfur R. Tecirlioglu and Poul Maddox-Hyttel for the innovative thinking, continuous support and hard work. We regard the establishment of the HMC as a fruit of this joint activity. The excellent technical support of many people, predominantly that of Anette Pedersen, Ruth Kristensen, Renee Hodgson and Klaus Villemoes, is also warmly acknowledged. We thank Maria A. Di Berardino, Cesare Galli, Torben Greve, Ronald H. F. Hunter, Irina Lagutina, Péter Zs. Nagy, Bjorn Oback, Teija T. Peura, László Rákosi, Tayfur R. Tecirlioglu and Matthew Wheeler for providing data, tehnical help and access to sources. This manuscript was the result of our joint efforts.


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