Posters
Presentation Type
Poster
Discipline Track
Biomedical Science
Abstract Type
Research/Clinical
Abstract
Background/Objective: We conducted this project to differentiate cell susceptibility to hypoxia (low O2 level) between immortalized Müller glia (MIO-M1) stored in -80 versus liquid nitrogen.
Method: MIO-M1 cells were cultured and seeded in 12 well plates at 40K/well and in 6 well plates at 75K/well until confluence. Cell culture conditions: centrifugation time and speed, neutralization, and storage temperature were varied. Cells were subjected to chemically induced hypoxia by treatment with 300 and 400uM CoCl2 for 24 and 48hours. Oxygen level was measured in cells, cells were imaged and counted.
Results: Centrifugation and neutralization conditions did not affect cell survival. We found that storing MIO-M1 cell in -80 makes these cells more susceptible to death by hypoxia. First, we found that %live MIO-M1 cells stored in liquid nitrogen is near 100% compared to ~60% live cells when stored in -80. Second, cells stored in -80 showed significant susceptibility to chemically induced hypoxia compared to cells stored in liquid nitrogen that survived treated for 24 and 48 hours.
Conclusion: We concluded that storage of these cells in liquid nitrogen maintain survival during hypoxic events. Cells’ susceptibility to hypoxia indicates that storage in -80 affect glycolysis efficiency in these cells to maintain survival. During hypoxia, mitochondrial is dysfunctional as they depend on O2 for ATP production during oxidative phosphorylation. Cells stored in liquid nitrogen survived hypoxic conditions, which may indicate that MIO-M1 depends on glycolysis for ATP production. Further investigation will proceed to determine Müller glia metabolism preference
Academic/Professional Position
Faculty
Mentor/PI Department
Molecular Science
Recommended Citation
Jassim, Assraa; Garcia, Martin; Tsin, Andrew; Valdez, Laura; and Martinez, Jose, "Storage Temperature Affects Müller Glia Susceptibility to Hypoxia" (2023). Research Symposium. 113.
https://scholarworks.utrgv.edu/somrs/2022/posters/113
Included in
Storage Temperature Affects Müller Glia Susceptibility to Hypoxia
Background/Objective: We conducted this project to differentiate cell susceptibility to hypoxia (low O2 level) between immortalized Müller glia (MIO-M1) stored in -80 versus liquid nitrogen.
Method: MIO-M1 cells were cultured and seeded in 12 well plates at 40K/well and in 6 well plates at 75K/well until confluence. Cell culture conditions: centrifugation time and speed, neutralization, and storage temperature were varied. Cells were subjected to chemically induced hypoxia by treatment with 300 and 400uM CoCl2 for 24 and 48hours. Oxygen level was measured in cells, cells were imaged and counted.
Results: Centrifugation and neutralization conditions did not affect cell survival. We found that storing MIO-M1 cell in -80 makes these cells more susceptible to death by hypoxia. First, we found that %live MIO-M1 cells stored in liquid nitrogen is near 100% compared to ~60% live cells when stored in -80. Second, cells stored in -80 showed significant susceptibility to chemically induced hypoxia compared to cells stored in liquid nitrogen that survived treated for 24 and 48 hours.
Conclusion: We concluded that storage of these cells in liquid nitrogen maintain survival during hypoxic events. Cells’ susceptibility to hypoxia indicates that storage in -80 affect glycolysis efficiency in these cells to maintain survival. During hypoxia, mitochondrial is dysfunctional as they depend on O2 for ATP production during oxidative phosphorylation. Cells stored in liquid nitrogen survived hypoxic conditions, which may indicate that MIO-M1 depends on glycolysis for ATP production. Further investigation will proceed to determine Müller glia metabolism preference