A flowchart of the Matrigel based method for transporting cells in semi-solid gel condition and at ambient temperature.

<p>A flowchart of the Matrigel based method for transporting cells in semi-solid gel condition and at ambient temperature.</p

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

<div><p>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.</p></div

Comparison of properties of cells recovered from medium supplemented Matrigel (MT) and thawed from cryopreserved vials in liquid nitrogen (LN).

<p>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 <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0128229#pone.0128229.g002" target="_blank">Fig 2</a> was used as a control. (a) Growth of MCF-7 cells treated with hormone (estradiol, <i>E2</i>) 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, **, <i>p <</i> 0.01; N.S., not significant.</p

The survival rate of various types of cells recovered from medium supplemented Matrigel and thawed from cryopreserved vials in liquid nitrogen.

<p>Various types of cells at 5×10⁶ per ml were cryopreserved in cryogenic vial in liquid nitrogen or stored in medium (34%) supplemented Matrigel (66%) at room temperature for 7 days, cells were then recovered for assays. (a) Cell survival rates were detected by Trypan blue exclusion assay. (b) Cell growth assay were done as described in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0128229#pone.0128229.g002" target="_blank">Fig 2(e)</a>. (c) and (d) Cell colony formation were determined as described in material and methods. The data are presented as the mean ± SD, *, <i>p <</i> 0.05; N.S., not significant.</p

The survival rate of MCF-7 cells stored in the cell growth medium supplemented Matrigel.

<p>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 *, <i>p <</i> 0.05; **, <i>p <</i> 0.01; n.s., not significant.</p

ncRDeathDB: A comprehensive bioinformatics resource for deciphering network organization of the ncRNA-mediated cell death system

<p>Programmed cell death (PCD) is a critical biological process involved in many important processes, and defects in PCD have been linked with numerous human diseases. In recent years, the protein architecture in different PCD subroutines has been explored, but our understanding of the global network organization of the noncoding RNA (ncRNA)-mediated cell death system is limited and ambiguous. Hence, we developed the comprehensive bioinformatics resource (ncRDeathDB, <a href="http://www.rna-society.org/ncrdeathdb" target="_blank">www.rna-society.org/ncrdeathdb</a>) to archive ncRNA-associated cell death interactions. The current version of ncRDeathDB documents a total of more than 4600 ncRNA-mediated PCD entries in 12 species. ncRDeathDB provides a user-friendly interface to query, browse and manipulate these ncRNA-associated cell death interactions. Furthermore, this resource will help to visualize and navigate current knowledge of the noncoding RNA component of cell death and autophagy, to uncover the generic organizing principles of ncRNA-associated cell death systems, and to generate valuable biological hypotheses.</p