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Daniel:Notebook/HiResChrPaint/2013-9-17
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=Fibroblast Adherence ([[Daniel:Notebook/HiResChrPaint/2013-9-9|Started 09/09/2013]])= [[Daniel:Notebook/HiResChrPaint|Back to Calendar]] ==Testing Cell Seeding Density 2 ([[Daniel:Notebook/HiResChrPaint/2013-9-16|From 9/16/2013]])== Since [[Daniel:Notebook/HiResChrPaint/2013-9-16|yesterday's]] experiment didn't give me the results I was looking for, I'm going to repeat the experiment over a longer time course. Further, I'm going to wait to take photos until at least 3 hours post incubation to give the cells plenty of growth time. Since I still have a few culture dishes left I'll be using those ===Protocol=== 1. Aspirate media and rinse with PBS 2. Add 500 uL trypsin and incubate 5 min. 3. Resuspend cells in 9.5 mL media 4. Add 100 uL cells to dishes 1 and 4 5. Add 50 uL cell suspension and 50 uL DMEM to dish 2 6. Spin down cells and resuspend in 250 uL trypsin + 4.75 mL DMEM; add 100 uL to dish 3 7. Timecourse at 3, 4.5 and 6 hours post initial incubation 8. Add 100 uL DMEM to dish 1** 3 hours post initial incubation <nowiki>**</nowiki>Note: This is changed from yesterday's protocol. When I looked at the dishes, dish 1 looked fairly weak and dish 4 looked good. Since I want to see if the kicker improves cell seeding/growth, I decided to give the 100uL DMEM to dish 1, to see if its seeding improved. Dish 1 and dish 4 were the same up to this point. ===Results=== The cells do not seem to be adhering well. Again. I'll leave them overnight, but it seems that this cell seeding exploration is moot, because even on the dishes it can be difficult to incubate these well. I'll have to think over the protocol. I must be doing something wrong that these fibroblasts do not want to grow.
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