This study was the first reported successful attempt to lyophilize nucleated somatic cells for downstream applications (Loi, et al. 2008). At the time, most efforts exploring potential applications for lyophilization were focused on long term storage of blood cells.
Sheep lymphocytes and granulosa cells were lyophilized with 50% and 40% nuclear integrity. Granulosa cells were then dispatched from the Institute of Animal Science at the Volcani Center in Israel to Teramo University in Italy by surface mail and stored in the dark at room temperature for three years before being used for nuclear transfer. The granulosa cells were then hydrated and immediately injected into enucleated oocytes.
Lyophilized somatic cells stored for 3 years at room temperature were able to direct embryonic development following injection into enucleated oocytes.
Results demonstrated that alternative systems for long-term storage of cell lines are possible, and would open unprecedented opportunities in the fields of biomedicine, bio-banking, and conservation strategies.
Freeze-Dried Somatic Cells Direct Embryonic Development after Nuclear Transfer
P. Loi, K. Matsukawa, G. Ptak, M. Clinton, J. Fulka Jr., Y. Natan, A. Arav
Plos One, vol. 3, e2978 (2008)
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Figure 1. Nuclear transfer with dried cells
Figure 2. Nuclear transfer with dried cells
Figure 3. Sheep embryos produced after nuclear transfer with dried leucocytes that were stored at room temperature for 3 years
|No. of oocytes injected||Survived (%)||Pronuclear like (%)||Cleavage (%)|
|359||320 (89.1)||285* (79.3)||—-|
|Cell donor||Oocyte injected (%)||Survived (%)||Cleavage (%)|
|Ram 3||45||40 (88.8%)||14 (2-4 cells) (31.1%)|
|Ram 2594||50||47 (94.0%)||28 (2-4-10 cells) (56.0%)|
|Ram 3089||39||36 (92.3%)||19 (2-4-10 cells) (48.7%)|
Table 1. Embryonic development after cloning using lyophilized sheep MNC as nuclear donors