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Transporting Cells in Semi-Solid Gel Condition and at Ambient Temperature.

Wang J, Chen P, Xu J, Zou JX, Wang H, Chen HW - PLoS ONE (2015)

Bottom Line: The hazardous nature of dry ice and liquid nitrogen, and the associated high shipping cost strongly limit their routine use.Our results showed that cells mixed with Matrigel at suitable ratios maintained excellent viability (>90%) for one week at room temperature and preserved the properties such as morphology, drug sensitivity and metabolism well, which was comparable to cells cryopreserved in liquid nitrogen.Upon arrival, it was found that over 90% of the cells were viable and grew well after replating.

View Article: PubMed Central - PubMed

Affiliation: Comprehensive Cancer Center, University of California Davis, Sacramento, California, 95817, United States of America; Department of Biochemistry and Molecular Medicine, School of Medicine, University of California Davis, Sacramento, California, 95817, United States of America.

ABSTRACT
Mammalian cells including human cancer cells are usually transported in cryovials on dry ice or in a liquid nitrogen vapor shipping vessel between different places at long distance. The hazardous nature of dry ice and liquid nitrogen, and the associated high shipping cost strongly limit their routine use. In this study, we tested the viability and properties of cells after being preserved or shipped over long distance in Matrigel mixture for different days. Our results showed that cells mixed with Matrigel at suitable ratios maintained excellent viability (>90%) for one week at room temperature and preserved the properties such as morphology, drug sensitivity and metabolism well, which was comparable to cells cryopreserved in liquid nitrogen. We also sent cells in the Matrigel mixture via FedEx service to different places at ambient temperature. Upon arrival, it was found that over 90% of the cells were viable and grew well after replating. These data collectively suggested that our Matrigel-based method was highly convenient for shipping live cells for long distances in semi-solid gel condition and at ambient temperature.

No MeSH data available.


Related in: MedlinePlus

The survival rate of MCF-7 cells stored in the cell growth medium supplemented Matrigel.Cells were preserved in cryogenic vials in liquid nitrogen (LN) for 14 days, and were thawed and used as a control for determined cell viability by trypan blue exclusion assay. (a) Images of cells stored in medium supplemented Matrigel in Eppendorf tubes. (b) Cells were recovered from medium (34%) supplemented Matrigel (66%) as described in Materials and Methods, the cell survival rate was determined at the indicated time. (c) Cells were preserved in the mixture with different ratios between medium and Matrigel for 7 days, and were recovered for determining cell survival rate. (d) Different densities of cells were preserved in medium (34%) supplemented Matrigel (66%) for 7 days, and were recovered for determining cell survival rate. (e) and (f) Cells were preserved in medium (34%) supplemented Matrigel (66%) for 7 days, and were recovered for cell growth assay as described in Materials and Methods. Representative images showed cellular morphology. The data are presented as the mean ± SD *, p < 0.05; **, p < 0.01; n.s., not significant.
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pone.0128229.g002: The survival rate of MCF-7 cells stored in the cell growth medium supplemented Matrigel.Cells were preserved in cryogenic vials in liquid nitrogen (LN) for 14 days, and were thawed and used as a control for determined cell viability by trypan blue exclusion assay. (a) Images of cells stored in medium supplemented Matrigel in Eppendorf tubes. (b) Cells were recovered from medium (34%) supplemented Matrigel (66%) as described in Materials and Methods, the cell survival rate was determined at the indicated time. (c) Cells were preserved in the mixture with different ratios between medium and Matrigel for 7 days, and were recovered for determining cell survival rate. (d) Different densities of cells were preserved in medium (34%) supplemented Matrigel (66%) for 7 days, and were recovered for determining cell survival rate. (e) and (f) Cells were preserved in medium (34%) supplemented Matrigel (66%) for 7 days, and were recovered for cell growth assay as described in Materials and Methods. Representative images showed cellular morphology. The data are presented as the mean ± SD *, p < 0.05; **, p < 0.01; n.s., not significant.

Mentions: In an experiment involving establishment of xenograft tumors, we accidentally found that leftover cells from injection, in the form of 50% Matrigel mixture, were highly viable after 4 days at room temperature. This observation prompted us to postulate that Matrigel might be useful in transporting cells in semi-solid gel condition at ambient temperatures. To test the hypothesis, MCF-7 breast cancer cells were resuspended in the mixture of 66% Matrigel and 34% cell culture medium in 1.5 ml-Eppendorf microcentrifuge tubes, and the mixture was allowed to gel at 37°C for two hours (Fig 2a). The tubes were left stand at bench at room temperature for different days, cells in the gel were then resuspended in complete cell growth medium to recover the cells (for details, see Materials and Methods). Cell viability was then determined by trypan blue exclusion assay. Remarkably, cells mixed with Matrigel and stored at room temperature maintained excellent viability for 1 week, which was comparable to cells cryopreserved in liquid nitrogen (Fig 2b). To identify the optimal conditions that gave the highest viability, different numbers of cells were mixed in different concentrations of Matrigel. The results showed that cell densities ranging from 1×106 to 5×106 cells per ml and Matrigel:medium ratios from 3:1 to 2:1 were optimal for cell viability (Fig 2c and 2d). Cell densities higher than 1×107 showed lower survival rate. Matrigel alone (i.e., undiluted, without medium) was not an optimal condition. Moreover, cells recovered from the Matrigel mixture displayed morphology and cell growth rates that were essentially identical to cells thawed from cryovials in liquid nitrogen (Fig 2e and 2f). These results demonstrate that the cell growth medium-supplemented Matrigel provides cells with a suitable survival environment for maintaining their viability at room temperature.


Transporting Cells in Semi-Solid Gel Condition and at Ambient Temperature.

Wang J, Chen P, Xu J, Zou JX, Wang H, Chen HW - PLoS ONE (2015)

The survival rate of MCF-7 cells stored in the cell growth medium supplemented Matrigel.Cells were preserved in cryogenic vials in liquid nitrogen (LN) for 14 days, and were thawed and used as a control for determined cell viability by trypan blue exclusion assay. (a) Images of cells stored in medium supplemented Matrigel in Eppendorf tubes. (b) Cells were recovered from medium (34%) supplemented Matrigel (66%) as described in Materials and Methods, the cell survival rate was determined at the indicated time. (c) Cells were preserved in the mixture with different ratios between medium and Matrigel for 7 days, and were recovered for determining cell survival rate. (d) Different densities of cells were preserved in medium (34%) supplemented Matrigel (66%) for 7 days, and were recovered for determining cell survival rate. (e) and (f) Cells were preserved in medium (34%) supplemented Matrigel (66%) for 7 days, and were recovered for cell growth assay as described in Materials and Methods. Representative images showed cellular morphology. The data are presented as the mean ± SD *, p < 0.05; **, p < 0.01; n.s., not significant.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4476595&req=5

pone.0128229.g002: The survival rate of MCF-7 cells stored in the cell growth medium supplemented Matrigel.Cells were preserved in cryogenic vials in liquid nitrogen (LN) for 14 days, and were thawed and used as a control for determined cell viability by trypan blue exclusion assay. (a) Images of cells stored in medium supplemented Matrigel in Eppendorf tubes. (b) Cells were recovered from medium (34%) supplemented Matrigel (66%) as described in Materials and Methods, the cell survival rate was determined at the indicated time. (c) Cells were preserved in the mixture with different ratios between medium and Matrigel for 7 days, and were recovered for determining cell survival rate. (d) Different densities of cells were preserved in medium (34%) supplemented Matrigel (66%) for 7 days, and were recovered for determining cell survival rate. (e) and (f) Cells were preserved in medium (34%) supplemented Matrigel (66%) for 7 days, and were recovered for cell growth assay as described in Materials and Methods. Representative images showed cellular morphology. The data are presented as the mean ± SD *, p < 0.05; **, p < 0.01; n.s., not significant.
Mentions: In an experiment involving establishment of xenograft tumors, we accidentally found that leftover cells from injection, in the form of 50% Matrigel mixture, were highly viable after 4 days at room temperature. This observation prompted us to postulate that Matrigel might be useful in transporting cells in semi-solid gel condition at ambient temperatures. To test the hypothesis, MCF-7 breast cancer cells were resuspended in the mixture of 66% Matrigel and 34% cell culture medium in 1.5 ml-Eppendorf microcentrifuge tubes, and the mixture was allowed to gel at 37°C for two hours (Fig 2a). The tubes were left stand at bench at room temperature for different days, cells in the gel were then resuspended in complete cell growth medium to recover the cells (for details, see Materials and Methods). Cell viability was then determined by trypan blue exclusion assay. Remarkably, cells mixed with Matrigel and stored at room temperature maintained excellent viability for 1 week, which was comparable to cells cryopreserved in liquid nitrogen (Fig 2b). To identify the optimal conditions that gave the highest viability, different numbers of cells were mixed in different concentrations of Matrigel. The results showed that cell densities ranging from 1×106 to 5×106 cells per ml and Matrigel:medium ratios from 3:1 to 2:1 were optimal for cell viability (Fig 2c and 2d). Cell densities higher than 1×107 showed lower survival rate. Matrigel alone (i.e., undiluted, without medium) was not an optimal condition. Moreover, cells recovered from the Matrigel mixture displayed morphology and cell growth rates that were essentially identical to cells thawed from cryovials in liquid nitrogen (Fig 2e and 2f). These results demonstrate that the cell growth medium-supplemented Matrigel provides cells with a suitable survival environment for maintaining their viability at room temperature.

Bottom Line: The hazardous nature of dry ice and liquid nitrogen, and the associated high shipping cost strongly limit their routine use.Our results showed that cells mixed with Matrigel at suitable ratios maintained excellent viability (>90%) for one week at room temperature and preserved the properties such as morphology, drug sensitivity and metabolism well, which was comparable to cells cryopreserved in liquid nitrogen.Upon arrival, it was found that over 90% of the cells were viable and grew well after replating.

View Article: PubMed Central - PubMed

Affiliation: Comprehensive Cancer Center, University of California Davis, Sacramento, California, 95817, United States of America; Department of Biochemistry and Molecular Medicine, School of Medicine, University of California Davis, Sacramento, California, 95817, United States of America.

ABSTRACT
Mammalian cells including human cancer cells are usually transported in cryovials on dry ice or in a liquid nitrogen vapor shipping vessel between different places at long distance. The hazardous nature of dry ice and liquid nitrogen, and the associated high shipping cost strongly limit their routine use. In this study, we tested the viability and properties of cells after being preserved or shipped over long distance in Matrigel mixture for different days. Our results showed that cells mixed with Matrigel at suitable ratios maintained excellent viability (>90%) for one week at room temperature and preserved the properties such as morphology, drug sensitivity and metabolism well, which was comparable to cells cryopreserved in liquid nitrogen. We also sent cells in the Matrigel mixture via FedEx service to different places at ambient temperature. Upon arrival, it was found that over 90% of the cells were viable and grew well after replating. These data collectively suggested that our Matrigel-based method was highly convenient for shipping live cells for long distances in semi-solid gel condition and at ambient temperature.

No MeSH data available.


Related in: MedlinePlus