Donor cells for nuclear transfer are usually prepared by the culture of fresh
tissue. However, animal carcasses are sometimes frozen without cryoprotectants and if
it were possible to obtain live cells from carcasses (tissue) preserved in this manner, it
could be very beneficial in nuclear transfer cloning of trophy or extinct animals.
This study tested the hypothesis that tissue samples of skin, muscle, and oral
mucosa could be cryopreserved without cryoprotection. The tissue samples were taken
from euthanized goats and placed into a -20°C freezer for varying lengths of time. The
samples were thawed by two different methods. One method was in 37°C water bath
and the other was on ice, thawing to room temperature from 1°C to 25°C. The samples
were then processed and placed into an incubator to evaluate cell growth.
Skin samples frozen for up to 34 days obtained cell growth to confluency and the
cells were then cryopreserved with cryoprotectant. The cells were able to tolerate the
potentially lethal effects of ice nucleation and dehydration brought about by ice
formation and colligative factors. Although this method of cryopreservation has been shown to yield growth of
cells that might be useful for nuclear transfer cloning, it is not the recommended method
to cryopreserve tissues if cryoprotectants are available or if only short term storage is
needed. These procedures would be especially useful when a precious animal dies
unexpectedly and cryoprotectant is not available and the sample can not be processed
before 10 days.
| Identifer | oai:union.ndltd.org:tamu.edu/oai:repository.tamu.edu:1969.1/ETD-TAMU-2819 |
| Date | 15 May 2009 |
| Creators | Charles, Lara Nicole |
| Contributors | Kraemer, Duane |
| Source Sets | Texas A and M University |
| Language | en_US |
| Detected Language | English |
| Type | Book, Thesis, Electronic Thesis, text |
| Format | electronic, application/pdf, born digital |
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