J Korean Acad Conserv Dent.  2009 Nov;34(6):491-499. 10.5395/JKACD.2009.34.6.491.

Comparison of viability of oral epithelial cells stored by different freezing methods

• Baek DY
• Lee SJ
• Jung HS
• Kim E

• KMID: 1986612
• DOI: http://doi.org/10.5395/JKACD.2009.34.6.491

Abstract

This study examined the influence of the storage methods on the viability of oral epithelial cells using conventional cell freezing storage, slow freezing preservation, rapid freezing preservation, and slow freezing preservation with a pressure of 2 Mpa or 3 Mpa. The cell viability was evaluated by cell counting, WST-1 and the clonogenic capacity after 6 days of freezing storage. After 6 days, the frozen cells were thawed rapidly, and the cell counting, WST-1, and clonogenic capacity values were measured and compared. 1. The results from cell counting demonstrated that conventional cryopreservation, slow freezing under a 2 Mpa pressure and slow freezing under a 3 Mpa pressure showed significantly higher values than slow freezing preservation and rapid freezing preservation (p<0.05). 2. The results from the optical density by WST-1 demonstrated that slow freezing under a 2 Mpa pressure showed significantly higher values than slow freezing preservation and rapid freezing preservation (p<0.05). 3. The clonogenic capacity demonstrated that slow freezing under a 2 Mpa pressure showed significantly higher values than slow freezing preservation and rapid freezing preservation (p<0.05).

Keyword

WST-1; Cell counting; Clonogenic capacity; Slow freezing under pressure; Slow freezing; Rapid freezing

MeSH Terms

Cell Count
Cell Survival
Cryopreservation
Epithelial Cells
Freezing

Figure

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  Figure 1 1℃/min freezing container "Mr. Frosty"

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  Figure 2 Schematic diagram of program freezer with pressure vessel. a. Oxygen container : 2,3 Mpa of pressure b. Program freezer c. Pressure bottle d. 2ml Cryotube: 1ml 65%RPMI+30%FBS+5%DMSO e. Cell suspension f. Pressure valve g. Thermometer

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  Figure 3 Cell counting by hemacytometer slide and trypan blue

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  Figure 4 Standard curve of WST-1 using monolayer epithelial cell(YD-38)

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  Figure 5 Clonogenic capacity of experimental groups

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  Figure 6 Schematic of physical events in cells during freezing.

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