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. 2021 Jan 22;13(3):402.
doi: 10.3390/cancers13030402.

Testing Lab-on-a-Chip Technology for Culturing Human Melanoma Cells under Simulated Microgravity

Dawid Przystupski  1   2 , Agata Górska  2   3 , Olga Michel  2 , Agnieszka Podwin  4 , Patrycja Śniadek  4 , Radosław Łapczyński  5 , Jolanta Saczko  2 , Julita Kulbacka  2
Affiliations

Testing Lab-on-a-Chip Technology for Culturing Human Melanoma Cells under Simulated Microgravity

Dawid Przystupski et al. Cancers (Basel). .

Abstract

The dynamic development of the space industry makes space flights more accessible and opens up new opportunities for biological research to better understand cell physiology under real microgravity. Whereas specialized studies in space remain out of our reach, preliminary experiments can be performed on Earth under simulated microgravity (sµg). Based on this concept, we used a 3D-clinostat (3D-C) to analyze the effect of short exposure to sµg on human keratinocytes HaCaT and melanoma cells A375 cultured on all-glass Lab-on-a-Chip (LOC). Our preliminary studies included viability evaluation, mitochondrial and caspase activity, and proliferation assay, enabling us to determine the effect of sµg on human cells. By comparing the results concerning cells cultured on LOCs and standard culture dishes, we were able to confirm the biocompatibility of all-glass LOCs and their potential application in microgravity research on selected human cell lines. Our studies revealed that HaCaT and A375 cells are susceptible to simulated microgravity; however, we observed an increased caspase activity and a decrease of proliferation in cancer cells cultured on LOCs in comparison to standard cell cultures. These results are an excellent basis to conduct further research on the possible application of LOCs systems in cancer research in space.

Keywords: LOC; cell death; cisplatin; melanoma; microgravity; multidrug resistance.

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Conflict of interest statement

The authors declare no conflict of interest.

Figures

Figure 1
Figure 1
Evaluation of HaCaT and A375 cells viability (A) and morphology (B) after 24 h culturing on various Lab-on-a-Chip (LOC) (ILM: inverted light microscope); scale bars: 50 μm. * Statistically significant differences in comparison to control cells (p ≤ 0.05).
Figure 2
Figure 2
Evaluation of HaCaT and A375 cells viability (A) and mitochondrial activity (B) after the exposure to simulated microgravity. Statistically significant differences in comparison to ctrl cells: *, ctrl + sμg cells: +, and LOC cells: # (p ≤ 0.05).
Figure 3
Figure 3
Caspase activity evaluation of HaCaT and A375 cells after the exposure to simulated microgravity; (RFU—relative fluorescence units). Statistically significant differences in comparison to ctrl cells: *, ctrl + sμg cells: +, and LOC cells: # (p ≤ 0.05).
Figure 4
Figure 4
Number of colonies (% of control cells) after the exposure to simulated microgravity and 7-day incubation of HaCaT and A375 cells (* p ≤ 0.05; data are presented as the mean amount of counted colonies). Statistically significant differences in comparison to ctrl cells: *, ctrl + sμg cells: +, and LOC cells: # (p ≤ 0.05).
Figure 5
Figure 5
The representative photographs of the morphology of HaCaT and A375 cells exposed to simulated microgravity (ILM); scale bars: 50 μm for HCS CellMask and 20 μm for F-actin.
Figure 6
Figure 6
Fabrication of all-glass LOCs (cross-section view), technology flow (A); the structure of the prepared LOCs (B).
Figure 7
Figure 7
The schematic representation of the procedure of simulated microgravity experiment.
See this image and copyright information in PMC

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