To prepare for cell seeding, you washed GBM cells (grown in T-75 flask) with RT

Question

To prepare for cell seeding, you washed GBM cells (grown in T-75 flask) with RT 1XPBS and trypsinized the cells with 1.5mL of 0.25% Trypsin-EDTA for 5 min @ 37 degree C. You then neutralized trypsin solution with 8.5mL of cell culture media. You transferred 30 mu L to a 1.5mL microcentrifuge tube, and mixed with 270 mu L trypan blue. You then transferred 10 mu L of the cell: trypan blue mixture to the hemocytometer and counted 950 cells from the five corner squares. Average # of cells DF # of cells per mu L # of cells per mL The total # of cells obtained from the cell cultute flask Dissolving 45 grams of glucose in 1,000 mL of water would yield what solution? How to make a 0.9% NaCl solution of IL in water?

Explanation / Answer

(a)Average number of cells = 950/5 = 190

(b)Dilution factor(DF)= 10^2

1.5 mL trypsin + 8.5 mL media = 10

30uL + 270 uL tryphan blue = 10

Total dilution factor = 10^2

(c) number of cells = average xDF x 10^4 cells/mL

=190 x 10^2 x 10^4 cells/mL

=190 x 10^6 cell/mL

(d) cells/uL = (190 x 10^6 cell/mL)/1000 = 190 x 10^3 cell/uL

(e) in 10 mL

Total number of cells = 190 x 10^7 cells

(9) 4.5%

45 g of glucose in 100 mL gives = 45%

45g of glucose in 1000 mL gives = 4.5%

(10) weigh 9 g of NaCl into 1000mL water = 0.9% NaCl

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