HEK 923 细胞的传代

HEK 923 subculture method in CELL Biology

1. prepare the devices in the hood, such as the pipette and flask(T-75), petri dishes(4undefined10)

2.prewarm the CDMEM, DPBS,TE(the combination solution by trysine and EDTA)

3.load the flask and dish with 8ml and 2 ml respectively and put them back

4. take the primary cells, rinsed them with 8ml DPBS

5. discard DPBS and rinse the cell with 2ml TE

6. wait and shake the flask for about 2 min until the detachment of cell occurred

7. add 8 ml CDMEM into the flask and transferred them to a centrifuge tube

8. centrifuge the cells at 40000rpm for 2 min

9. discard the upper solution and add 2ml CDMEM

10. seperate the cell with 600 ul pipette

11. transfered 100ul(one day) or 25ul(one week) into a new flask for culture. or transfer 40 ul cell to the dishes for another transfection.

I Just do the experiment in this afternoon and i will do the transfection tomorrow. So we can discuss more on BBS