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

Do-Young Baek; Seung-Jong Lee; Han-Sung Jung; EuiSeong Kim

doi:10.5395/JKACD.2009.34.6.491

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Original Article Comparison of viability of oral epithelial cells stored by different freezing methods: Do-Young Baek, Seung-Jong Lee, Han-Sung Jung, EuiSeong Kim; Journal of Korean Academy of Conservative Dentistry 2009;34(6):491-499.
DOI: https://doi.org/10.5395/JKACD.2009.34.6.491
Published online: November 30, 2009

Department of Conservative Dentistry, College of Dentistry, Yonsei University, Korea.

Corresponding Author: Euiseong Kim. Department of Conservative Dentistry, College of Dentistry, Yonsei University, 134 Shinchon-dong, Sudaemoon-gu, Seoul, Korea. Tel: 82-2-2228-3151, Fax: 82-2-313-7575, andyendo@yuhs.ac

• Received: September 1, 2009 • Revised: September 25, 2009 • Accepted: October 6, 2009

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Abstract
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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).
Keywords: WST-1; Cell counting; Clonogenic capacity; Slow freezing under pressure; Slow freezing; Rapid freezing

REFERENCES

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Figure 1

1℃/min freezing container "Mr. Frosty"

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

Figure 3

Cell counting by hemacytometer slide and trypan blue

Figure 4

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

Figure 5

Clonogenic capacity of experimental groups

Figure 6

Schematic of physical events in cells during freezing.

Table 1

The averages and standard deviations of viable cell number (logN)

a,b: Different letters denote statistically significant (p<0.05)

Table 2

The Median and IQR of optical density of WST-1

a,b,c: Different letters denote statistically significant (p<0.05)

Table 3

The Median and IQR of the number of colonies (number of cells seeded : 1×10³)

a,b,c: Different letters denote statistically significant (p<0.05)

Tables & Figures

REFERENCES

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

J Korean Acad Conserv Dent. 2009;34(6):491-499. Published online November 30, 2009

DOI: https://doi.org/10.5395/JKACD.2009.34.6.491

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Figure

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

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

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

Figure 3 Cell counting by hemacytometer slide and trypan blue

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

Figure 5 Clonogenic capacity of experimental groups

Figure 6 Schematic of physical events in cells during freezing.

Figure 1

Figure 2

Figure 3

Figure 4

Figure 5

Figure 6

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

Table 1

The averages and standard deviations of viable cell number (logN)

a,b: Different letters denote statistically significant (p<0.05)

Table 2

The Median and IQR of optical density of WST-1

a,b,c: Different letters denote statistically significant (p<0.05)

Table 3

The Median and IQR of the number of colonies (number of cells seeded : 1×10³)

a,b,c: Different letters denote statistically significant (p<0.05)

Table 1 The averages and standard deviations of viable cell number (logN)

a,b: Different letters denote statistically significant (p<0.05)

Table 2 The Median and IQR of optical density of WST-1

a,b,c: Different letters denote statistically significant (p<0.05)

Table 3 The Median and IQR of the number of colonies (number of cells seeded : 1×103)

a,b,c: Different letters denote statistically significant (p<0.05)