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Investigation of Murine T-Cells and Cancer Cells under Thermal Stressors and 2D Slow Rotating System Effects as a Testbed for Suborbital Flights

Pedro J. Llanos
Pedro J. Llanos
 y 
Kristina Andrijauskaite
Kristina Andrijauskaite
   | 17 sept 2019
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Article Category: Research Note
Publicado en línea: 17 sept 2019
Páginas: 45 - 61
DOI: https://doi.org/10.2478/gsr-2019-0006
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© 2019 Pedro J. Llanos, Kristina Andrijauskaite, published by Sciendo
This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 License.

Figure 1

Phase contrast microscopy. Evolution of T-cells after activation. Representative pictures of microscopic fields of each condition from two independent experiments. (A) and (B) Not activated cells (unstimulated). (C) Stimulated T-cells (primed with cytokine IL-2) after 2 days of activation. (E), (F), and (G) Zoomed sections of (C) as indicated by the solid, dotted, and dashed lines enclosing each boxed section. (H) Stimulated T-cells (not primed with cytokine). (I) Stimulated T-cells with IL-2. (J) Zoomed sections of (I) after 2 days of activation. A, B, C: 4× magnification, E, F, H, I: 10× magnification, J: 40× magnification; bar 100 mm.
Phase contrast microscopy. Evolution of T-cells after activation. Representative pictures of microscopic fields of each condition from two independent experiments. (A) and (B) Not activated cells (unstimulated). (C) Stimulated T-cells (primed with cytokine IL-2) after 2 days of activation. (E), (F), and (G) Zoomed sections of (C) as indicated by the solid, dotted, and dashed lines enclosing each boxed section. (H) Stimulated T-cells (not primed with cytokine). (I) Stimulated T-cells with IL-2. (J) Zoomed sections of (I) after 2 days of activation. A, B, C: 4× magnification, E, F, H, I: 10× magnification, J: 40× magnification; bar 100 mm.

Figure 2

Survival rate of T-cells exposed to various temperatures for the indicated time (in hours). (A) T-cells with no cytokines (single experiment). (B) Lung cancer cells (LLC) (single experiment). (C) T-cells primed with IL-2 cytokine and compared with control (single experiment). (D) T-cells primed with IL-12 cytokines and compared with control (single experiment). (E) Color differentiation of lung cancer cells.
Survival rate of T-cells exposed to various temperatures for the indicated time (in hours). (A) T-cells with no cytokines (single experiment). (B) Lung cancer cells (LLC) (single experiment). (C) T-cells primed with IL-2 cytokine and compared with control (single experiment). (D) T-cells primed with IL-12 cytokines and compared with control (single experiment). (E) Color differentiation of lung cancer cells.

Figure 3

T-cells aggregates formation after being exposed to a slow rotation system for 3 days. (A) Cell structures from L to R: spleen 1 with IL-2 activation, spleen 2 with IL-2 activation, spleen 2 with no IL-2 activation, spleen 3 with IL-2 activation, spleen 3 with no activation, 4× magnification. (B) four tubes (control, IL-2, IL-12, IL-2/IL-12); Control (center); IL-2 (right). Zoomed sections of control and IL-2 are five times larger as shown in the far right. (C) IL-12. Zoomed section is twice larger; comparison of IL-2 and IL-12. Zoomed sections are four times larger. Microscopy images represent two independent experiments.
T-cells aggregates formation after being exposed to a slow rotation system for 3 days. (A) Cell structures from L to R: spleen 1 with IL-2 activation, spleen 2 with IL-2 activation, spleen 2 with no IL-2 activation, spleen 3 with IL-2 activation, spleen 3 with no activation, 4× magnification. (B) four tubes (control, IL-2, IL-12, IL-2/IL-12); Control (center); IL-2 (right). Zoomed sections of control and IL-2 are five times larger as shown in the far right. (C) IL-12. Zoomed section is twice larger; comparison of IL-2 and IL-12. Zoomed sections are four times larger. Microscopy images represent two independent experiments.

Figure 4

Formation of T-cell aggregates in a slow rotation system after 3 days. (A). Aggregates of cells in IL-2. (B) Cells in IL-12. (C) Cells in IL-2/IL-12. (D) Early stages (+1 days) of cells in IL-12 forming swirling-like aggregates or frond-like structures ~3 mm long. (E), (F), and (G)–(H) are pictures of T-cells clusters for IL-2, IL-12, and IL-2/IL-12, respectively at 4× magnification. Microscopy images represent one independent experiment.
Formation of T-cell aggregates in a slow rotation system after 3 days. (A). Aggregates of cells in IL-2. (B) Cells in IL-12. (C) Cells in IL-2/IL-12. (D) Early stages (+1 days) of cells in IL-12 forming swirling-like aggregates or frond-like structures ~3 mm long. (E), (F), and (G)–(H) are pictures of T-cells clusters for IL-2, IL-12, and IL-2/IL-12, respectively at 4× magnification. Microscopy images represent one independent experiment.

Figure 5

Formation of aggregates after 116 h of slow rotation system operation at 4× magnification. (A) Three large cell structures with IL-2; zoom of each of the three large structures shown in first left picture of A. (B) Cell structures in control (first two pictures); cell structures in IL-12 (third picture); cell structures in IL-2/IL-12 (last picture). Microscopy images represent one independent experiment.
Formation of aggregates after 116 h of slow rotation system operation at 4× magnification. (A) Three large cell structures with IL-2; zoom of each of the three large structures shown in first left picture of A. (B) Cell structures in control (first two pictures); cell structures in IL-12 (third picture); cell structures in IL-2/IL-12 (last picture). Microscopy images represent one independent experiment.

Figure 6

T-cells counts (with standard errors) during slow rotation system study after 140 h obtained from two separate sets of cells from two independent experiments.
T-cells counts (with standard errors) during slow rotation system study after 140 h obtained from two separate sets of cells from two independent experiments.

Figure 7

Cell aggregates visualized under the microscope at original magnification from one independent experiment. (A) Control. (B) IL-2. (C) IL-12. (D) IL-2 and IL-12.
Cell aggregates visualized under the microscope at original magnification from one independent experiment. (A) Control. (B) IL-2. (C) IL-12. (D) IL-2 and IL-12.

Figure 8

(A) IL-12 and IL-2/IL-12 on the left picture where the filament is seen on Day 6. Different colors observed across different conditions. (B) Zoom of some aggregates showing various geometries induced by clinostat effects for IL-2, IL-12, IL-2/IL-12, and control conditions. Sections circled in red show aggregates for different conditions. Images represent one independent experiment at original magnification.
(A) IL-12 and IL-2/IL-12 on the left picture where the filament is seen on Day 6. Different colors observed across different conditions. (B) Zoom of some aggregates showing various geometries induced by clinostat effects for IL-2, IL-12, IL-2/IL-12, and control conditions. Sections circled in red show aggregates for different conditions. Images represent one independent experiment at original magnification.

Figure 9

Survival rate of cancer cells exposed to different temperatures for a time span of 130 h. (A) MCF7 breast cancer cells (single experiment). (B) Colon cancer cells (single experiment). Control cells at 37℃ have no error since the other sets are compared with this set. This should be from two independent experiments and should be averages.
Survival rate of cancer cells exposed to different temperatures for a time span of 130 h. (A) MCF7 breast cancer cells (single experiment). (B) Colon cancer cells (single experiment). Control cells at 37℃ have no error since the other sets are compared with this set. This should be from two independent experiments and should be averages.

Figure 10

Experimental design diagram outlining the description and source of T-cells and cancer cells with conducted experimental procedures and outcomes.
Experimental design diagram outlining the description and source of T-cells and cancer cells with conducted experimental procedures and outcomes.
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