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Bacteria

Kirkstall and Professor Robert Poole at the University of Sheffield have recently concluded a short project, funded by the BBSRC Metals in Biology NIBB. Its aims were:

  1. Establishment of bacterial (Escherichia coli) growth in minimal media in Quasi Vivo®.
  2. Determination of absorption/sequestering of metals from the cell-free media by Quasi Vivo® chamber materials (silicone and acrylic).

During the project, we showed that viability of E. coli grown in Quasi Vivo® is not significantly different from those grown in standard glass vessels (Figure 1).

We also showed that that the chambers do not abstract metals from the growth medium, nor leach metals into cell-free media (Table 1). This applies even when the medium has been treated to deplete selected metals thus providing a potential concentration gradient from the Quasi Vivo® chamber materials (silicone and acrylic) to the medium.

We conclude that these vessels are suitable for growths involving metal-controlled conditions.

Figure 1. Growth curve of E. coli MG1665 in Quasi Vivo® systems. Quasi Vivo: QV 600 vessel; positive Control, glass tubes; negative control: DMM in Quasi Vivo® vessel without bacterial culture.

Metals detected in uninoculated medium after incubation in vessel (mg/ml) ND = below detection limits of ICP-MS analysis
Vessel Metal Fe-limited Zn-limited Mn-limited
24 48 72 24 48 72 24 48 72
QV600 Fe ND ND ND 0.01 0.03 0.05 0.06 0.09 0.13
Zn 0.06 0.18 0.22 ND ND ND 0.002 0.003 0.01
QV900 Fe ND ND ND ND 0.02 ND ND 0.04 0.05
Zn 0.02 0.04 0.03 ND ND ND ND ND ND

Table 1. ICP-MS analysis of various metal-limited media after incubation in Quasi Vivo® chambers.

We conclude that these vessels are suitable for growths involving metal-controlled conditions. Thus Kirkstall will be expanding it’s research to work on models combining human cells and bacteria.

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