Mouse cloning using a drop of peripheral blood.

Area of Science:

• Reproductive Biology
• Animal Genetics
• Cellular Biology

Background:

• Somatic cell nuclear transfer (SCNT) is a key technology for producing cloned animals.
• Noninvasive collection of donor cells is highly desirable for practical SCNT applications.
• The feasibility of using peripheral blood cells for SCNT in living animals has not been fully established.

Purpose of the Study:

• To investigate the efficacy of using peripheral blood cells from living mice for SCNT.
• To determine if different leukocyte subpopulations (granulocytes/monocytes and lymphocytes) yield varying SCNT success rates.
• To assess the potential of this method for cloning genetically modified and inbred mouse strains.

Main Methods:

• Collection of small blood samples (15-45 μl) from the tail vein of donor mice.
• Preparation of nucleated cell suspensions by lysing red blood cells.
• SCNT using randomly selected leukocyte nuclei, followed by cloning of specific leukocyte types (granulocytes/monocytes and lymphocytes) and application to different mouse strains.

Main Results:

• Cloned offspring were successfully produced from peripheral blood leukocytes with a 2.8% birth rate.
• Granulocytes/monocytes and lymphocytes yielded comparable birth rates (2.1% and 1.7%, respectively).
• Normal-looking offspring were obtained from four different genetically modified and inbred strains using granulocyte/monocyte cloning.

Conclusions:

• Peripheral blood cells from living mice are a viable source for SCNT, enabling the noninvasive generation of cloned mice.
• Granulocyte/monocyte cloning is a promising approach, particularly for preserving valuable genetic resources from modified or inbred strains.
• This SCNT strategy offers a practical method for rescuing infertile or 'last-of-line' animals.