Figure 2
From: Rapid adaptation to microgravity in mammalian macrophage cells
Timeline of the flight (1–12) and ground ref. (13) experiments and data analysis. (1) NR8383 macrophageal cells (4.2 ml, 2.7 × 106 cells/ml) were filled in stock culture bags (SCBs). Cells were stored at −80 °C before, during and after upload to the International Space Station (ISS). Chemicals (luminol (Lum), zymosan (Zym), horse radish peroxidase (HRP)) were prepared, tested for their reactivity, and filled into extra liquid bags (ELBs) (3.2 ml) which were stored at −80 °C before and during the upload to the ISS. On board the ISS ELBs were stored at −20 °C. (2) The two sterile handling mechanism plate units (HMPU) were prefilled before upload. The internal filling bags (IFB) 1 and 2 were filled with 1.5 ml borate buffer and 2.27 ml NaCl, respectively. The four measurement bags (MB1–4) were filled with 2.4 ml of cell culture medium (ccm) each and the tubings with 0.2 ml ccm. The two HMPUs were stored before, during and after upload at 4 °C. (3) The experiment on board the ISS started with pre-warming the BIOLAB Rack to 37 °C. Thereafter, the pre-filled HMPU was integrated into the BIOLAB centrifuge. (4) ELBs were thawed and mixed. (5) The ELBs were implemented into the Automatic Ambient Stowage-Insert (AAS-I). (6) NR8383 cells in the SCB were thawed and mixed. (7) The SCB was integrated into the designated space in the HMPU. Liquids were transferred by the automated handling mechanism (HM) from the AAS-I into IFB1 and IFB2 and from the SCB into MB1-4. (8) Centrifugation at 37 °C for 3 h for cell recovery. (9) Turning-on of the 4 Photomultiplier Tubes (PMTs) and start of the measurement acquisition, experimental centrifugation regimes, and (10) pumping of the liquids from IFB1 and IFB2 into MB1-4 to start the oxidative burst reaction. After injection of the fluids from IFB1 and IFB2, the concentrations in MB1-4 were 0.44 × 106 NR8383 cells per ml, 1.7 mM luminol, 1.1 mg/ml zymosan and 55.85 U/ml HRP. (11) Live data downlink during the experiment. (12) Post flight data analysis. (13) The ground reference experiments (GRE1 and GRE2) were conducted in the BIOLAB engineering model (EM) following the same procedure as the flight experiments (Steps 1–12), except that there was no centrifugation in Steps 8 and 9. The cells recovered for 3 h under Earth 1 g conditions at 37 °C.
