Presentation on theme: "Cell Division in Microgravity"— Presentation transcript:
1Cell Division in Microgravity By Natasha Garamani, Jennifer Jiang, Jasmine Kuo, Kara Lukas, Elyssia Widjaja
2Background Cell division is an integral component of life. All organisms must go through the cell cycle in order to grow, develop, and reproduce.Past experiments that have been conducted on Earth in simulated microgravity have shown that the lack of gravity causes cells to divide at a slower rate.
3Purpose To discover the effect of microgravity on cell division To compare the rates of cell division in the absence and presence of gravity
4HypothesisIf cells are exposed to microgravity during their life cycles, then there will be a reduction in cell division because ofdisorganization of microtubuleschange in actin filamentschromatin less denseincreased apoptosis1. Microtubules are necessary for cytokinesis (mitotic spindle) 2. Microfilaments (cleavage furrow) 3. Chromatin condensation is essential in formation of chromosomes. 4. Programmed cell death
5Materials Fibroblasts Eagle’s Minimum Essential Medium (EMEM) 1% Fetal Bovine Serum (FBS)Light MicroscopeHemocytometerPipetteTrypan Blue1. They play a critical role in the healing of wounds. 2. EMEM helps with the preparation of cells 3. FBS provides the essential nutrients for the cells. 4. LM used to observe cells. 5. Hemocytometer is used to count cells. 6. Pipette used to transfer cells. 7. Trypan Blue to stain the cells so we can see them (counting purposes)
6Procedures - Ground Experiment Preparation of flasksCoat inside of flasks with 3 mL of poly-lysine solution for 5 minutesAspirate the solution and let it dry overnightTransfer 500 fibroblast cells into the flask with a pipetteAdd 5 mL of EMEM with 1% FBSIncubate cells overnightTo improve cell adherence to the surface of the ampoule
7Procedures - Ground Experiment 2. Initiation of ExperimentAdd 6.3 EMEM mixed with 1% FBS to both flasksKeep one flask in the incubator and put the other flask into a controlled room temperature environmentAllow the cells to continue to grow and divide for 12 days
8Procedures - Ground Experiment 3. Counting the CellsWash the cells with 2 ml of Phosphate-Buffered Saline solutionAdd 0.5 ml trypsin/EDTA to cause disassociation of the cells from the flask and rotate for 1 minute.Allow the cells to detach and add 1 ml of PBS.1. wash the cells and remove the serum which will inhibit the later use of trypsin. 6. Place 10 μL of the cells in the hemocytometer. Then count the cells in the central gridded square.Take that number and multiply by 104 to estimate the number of cells per ml. Repeat this procedure several times to ensure accuracy and average the count. The cell culture from each experiment will be mixed with the dye Trypan blue, which will be present only in dead cells and excluded in viable cells; we will count both dead and viable cells. After the calculation, we will compare the amount of cells produced as a result of each experiment
9Procedures 3. Counting the Cells (continued) Remove medium from the flask, place it in a 15 ml centrifuge tube, and spin for 5 minutes in a tabletop centrifuge to pellet the cells.Resuspend the pellet in 0.5 ml of PBSStain culture sample with trypan blueCount the cells collected from culture using a hemocytometer.2. Using hemocytometer
12ConclusionGrowth is more prominent for cells in incubator (at 37 degrees Celsius)Growth better in incubator b/c mimics body temperature
13Acknowledgements Mr. Wyeth Collo for his guidance as our advisor. Dr. Susan Kane and Ms. Erin Denny of City of Hope Cancer Center, Developmental Cancer Therapeutics Program, Comprehensive Cancer Center for providing the lab space and materials.Smithsonian Air and Space Museum for hosting this event.SSEP for this invaluable experience.