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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

Comparison of properties of cells recovered from medium supplemented Matrigel (MT) and thawed from cryopreserved vials in liquid nitrogen (LN).MCF-7 cells were preserved in medium (34%) supplemented Matrigel (66%) for 7 days, and were recovered as described in Materials and Methods. Cryopeserved cells in liquid nitrogen as Fig 2 was used as a control. (a) Growth of MCF-7 cells treated with hormone (estradiol, E2) or vehicle was assayed. (b) Glucose consumption and lactate production were detected as described in material and methods. (c) Cell growth was detected by MTT assay after cells treated with chemotherapy drug doxorubicin and taxol for 48 hours. The data are presented as the mean ± SD, **, p < 0.01; N.S., not significant.
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pone.0128229.g003: Comparison of properties of cells recovered from medium supplemented Matrigel (MT) and thawed from cryopreserved vials in liquid nitrogen (LN).MCF-7 cells were preserved in medium (34%) supplemented Matrigel (66%) for 7 days, and were recovered as described in Materials and Methods. Cryopeserved cells in liquid nitrogen as Fig 2 was used as a control. (a) Growth of MCF-7 cells treated with hormone (estradiol, E2) or vehicle was assayed. (b) Glucose consumption and lactate production were detected as described in material and methods. (c) Cell growth was detected by MTT assay after cells treated with chemotherapy drug doxorubicin and taxol for 48 hours. The data are presented as the mean ± SD, **, p < 0.01; N.S., not significant.

Mentions: We next performed experiments to further characterize cells recovered from seven days of storage in the medium-supplemented Matrigel. MCF-7 cells are estrogen receptor-positive breast cancer cells and highly sensitive to estrogen in growth stimulation. Upon recovery, cells from the medium-supplemented Matrigel and from the cryovials showed essentially identical slow growth curves in the absence of estrogen and faster growth curves with estrogen (Fig 3a). Aberrant metabolism is a hallmark of cancer [10]. Tumor cells display a heightened glucose uptake, glycolysis, and lactate production. Cells recovered from the medium-supplemented Matrigel and from cryostorage showed the similar a glucose uptake and lactate production (Fig 3b). MCF-7 cells are also sensitive to growth inhibition by chemotherapeutic drugs taxol and doxorubicin [11,12]. Cells recovered from the medium-supplemented Matrigel and from the cryovials were growth-inhibited in an identical manner by doxorubicin and taxol (Fig 3c). These experiments illustrated that the method of Matrigel-based transporting of cells can preserve the cancer cell properties well.


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)

Comparison of properties of cells recovered from medium supplemented Matrigel (MT) and thawed from cryopreserved vials in liquid nitrogen (LN).MCF-7 cells were preserved in medium (34%) supplemented Matrigel (66%) for 7 days, and were recovered as described in Materials and Methods. Cryopeserved cells in liquid nitrogen as Fig 2 was used as a control. (a) Growth of MCF-7 cells treated with hormone (estradiol, E2) or vehicle was assayed. (b) Glucose consumption and lactate production were detected as described in material and methods. (c) Cell growth was detected by MTT assay after cells treated with chemotherapy drug doxorubicin and taxol for 48 hours. The data are presented as the mean ± SD, **, 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.g003: Comparison of properties of cells recovered from medium supplemented Matrigel (MT) and thawed from cryopreserved vials in liquid nitrogen (LN).MCF-7 cells were preserved in medium (34%) supplemented Matrigel (66%) for 7 days, and were recovered as described in Materials and Methods. Cryopeserved cells in liquid nitrogen as Fig 2 was used as a control. (a) Growth of MCF-7 cells treated with hormone (estradiol, E2) or vehicle was assayed. (b) Glucose consumption and lactate production were detected as described in material and methods. (c) Cell growth was detected by MTT assay after cells treated with chemotherapy drug doxorubicin and taxol for 48 hours. The data are presented as the mean ± SD, **, p < 0.01; N.S., not significant.
Mentions: We next performed experiments to further characterize cells recovered from seven days of storage in the medium-supplemented Matrigel. MCF-7 cells are estrogen receptor-positive breast cancer cells and highly sensitive to estrogen in growth stimulation. Upon recovery, cells from the medium-supplemented Matrigel and from the cryovials showed essentially identical slow growth curves in the absence of estrogen and faster growth curves with estrogen (Fig 3a). Aberrant metabolism is a hallmark of cancer [10]. Tumor cells display a heightened glucose uptake, glycolysis, and lactate production. Cells recovered from the medium-supplemented Matrigel and from cryostorage showed the similar a glucose uptake and lactate production (Fig 3b). MCF-7 cells are also sensitive to growth inhibition by chemotherapeutic drugs taxol and doxorubicin [11,12]. Cells recovered from the medium-supplemented Matrigel and from the cryovials were growth-inhibited in an identical manner by doxorubicin and taxol (Fig 3c). These experiments illustrated that the method of Matrigel-based transporting of cells can preserve the cancer cell properties well.

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