Experiments for drawing growth curves of cultured cells

Summary

Cell growth curves are an important means of understanding the detailed process of proliferation of cultured cells and the basic laws of cell growth.

Principle

The basic principle of drawing the growth curve of cultured cells is that by continuously counting the cultured cells, the growth curve of the cells can be drawn, so as to determine whether the growth of the cultured cells is stable or not, the process of the change of the cell proliferation rate and the time of the peak of the proliferation, so as to provide the optimal time for the cell generation, freezing, and the further use of the cultured cells for scientific experiments.

Operation method

Experiments for drawing growth curves of cultured cells

Principle

The basic principle of drawing the growth curve of cultured cells is that by continuously counting the cultured cells, the growth curve of the cells can be drawn, so as to determine whether the growth of the cultured cells is stable or not, the process of the change of the cell proliferation rate and the time of the peak of the proliferation, so as to provide the optimal time for the cell generation, freezing, and the further use of the cultured cells for scientific experiments.

Materials and Instruments

Equipment:
① Cells in adherent wall culture
② 24-well culture plate
③ Pipette
③ Suction tube ④ Gel cap
⑤ Centrifuge tube
⑥ Petri dish
⑦ Semi-logarithmic coordinate paper
⑧ Coverslip
⑨ Cell counter plate
⑩ Centrifuge
⑪ Inverted microscope
⑫ General optical microscope
Ultra-clean table
⑭ CO
2
Incubator
Reagents
① D-Hanks liquid
① D-Hanks liquid ② DMEM medium (including 10% calf serum)
③ 0.25% trypsin digestive solution

Move

The basic procedure for plotting the growth curve of cultured cells can be divided into the following steps:
A. Digest monolayer cultured cells in logarithmic growth phase with 0.25% trypsin solution and collect the digested cells in a 10 mL centrifuge tube.
B. Centrifuge the cells for 5 min at 800~1000 r/min and discard the supernatant.
C. Prepare a single-cell suspension by adding DMEM medium (containing 10% calf serum).
D. Blow the cells evenly and count them. E. Inoculate 24-well culture plate with 2 x 104~5 x 104 cells/mL. Blow the cells well and count them.
E. Inoculate the cells into 24-well plates at a concentration of 2 x ^104-5 x ¹⁰⁴ cells/mL.
F. Digest the cells in 3 wells with trypsin solution every day, dilute them with D-hanks solution, and count them under the microscope.
G. Calculate the average of the cell counts of the 3-well cultures.
H. Digest the 3 wells of the cells as described above for 7 consecutive days and count the cells. average value.

Caveat

1. The number of cells inoculated into the culture plate wells should be consistent in each well. The amount of inoculum should be appropriate, not too little or too much, too little will make the cell growth cycle prolonged, too much will lead to the cells before the completion of the experiment that need to be passed on, in both cases the growth curve can not accurately reflect the growth of the cells.

For more product details, please visit Aladdin Scientific website.

https://www.aladdinsci.com/