Freeze-Dried Somatic Cells Direct Embryonic Development after Nuclear Transfer

The natural capacity of simple organisms to survive in a dehydrated state has long been exploited by man, with lyophylization the method of choice for the long term storage of bacterial and yeast cells. More recently, attempts have been made to apply this procedure to the long term storage of blood cells. However, despite significant progress, practical application in a clinical setting is still some way off. Conversely, to date there are no reports of attempts to lyophilize nucleated somatic cells for possible downstream applications. Here we demonstrate that lyophilised somatic cells stored for 3 years at room temperature are able to direct embryonic development following injection into enucleated oocytes. These remarkable results demonstrate that alternative systems for the long-term storage of cell lines are now possible, and open unprecedented opportunities in the fields of biomedicine and for conservation strategies

Total cell number and ICM and trophectoderm cell counting in control, and freeze dried cells-derived blastocysts.

a-b<p>differ significantly for p<0.05</p

In vitro development of enucleated oocytes injected with freeze-dried and fresh, control granulosa cells.

<p>The culture were maintained for 7–8 day in medium SOF plus amino-acids and BSA, with FCS added a day 4. reconstructed embryos were checked every 24 hours for development.</p><p>Test χ²</p>a<p>P = 0.88;</p>b<p>P = 0.17;</p>c<p>P = 0.56</p

g–h, re-hydrated granulosa cells Propidium Iodine staining (g, bright field; h, epifluorescence) ×150.

<p>i, nuclear transfer of freeze-dried granulosa cell×60. j, blastocyst produced from nuclear transfer of freeze dried granulosa cells×40.</p