Method for evaluating adherence strength of cells by utilizing cell detachment agent
A cell and detachment technology, applied in the field of measurement, can solve the problems of difficult automation, high threshold of instruments and technology, cumbersome operation, etc., and achieve the effect of reducing experimental time, improving measurement accuracy and result parallelism, and simplifying experimental steps
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Embodiment 1
[0021] Embodiment 1: Using the method of the present invention to detect cell adhesion strength
[0022] Step 1: Prepare the human osteosarcoma cell line MG-63 at 4 x 10 4 piece / cm 2 Cells were seeded in a 96-well cell culture plate at a density of 1.4×10 4 50 wells were inoculated, and 200 μl of medium was added to each well. Place in a 37°C cell culture incubator for 24 hours to grow to a confluence of 80%;
[0023] Step 2: using trypsin solution as the cell detachment reagent, the cell detachment reagent is half-diluted to obtain a set of detachment reagent solution concentrations, and the dilution times of the concentration gradients of each detachment reagent solution are respectively 2 0 ,2 1 ,2 2 ,2 3 ,2 4 ,2 5 ,2 6 The concentration of each gradient solution is 50%, 25%, 12.5%, 6.25%, 3.13%, 1.57%, 0.79% of the standard solution concentration respectively;
[0024] Step 3: Aspirate the culture medium of 45 wells in the 96-well plate of the dry cell culture pl...
Embodiment 2
[0036] Embodiment 2: Using the method of the present invention to detect cell adhesion strength
[0037] 1. Prepare the human osteosarcoma cell line MG-63 at 4×10 4 piece / cm 2 Cells were seeded in 96-well plates at a density of 1.4×10 4 50 wells were inoculated, and 200 μl of medium was added to each well. Place in a 37°C cell culture incubator for 24 hours to grow to a confluence of 80%;
[0038] 2. Take the EDTA solution as the standard solution, dilute it twice and a half, and prepare a concentration gradient solution. The concentration of each gradient solution is 50%, 25%, 12.5%, 6.25%, 3.13%, 1.57%, and 0.79% of the concentration of the standard solution;
[0039] 3. Drain the medium in 45 wells of the 96-well plate, and treat the cells with PBS and EDTA concentration gradient solution (0.79%-100% standard solution concentration), 100 μl per well, and make 5 replicates for each concentration. The last 5 wells with culture medium were used as the control of cell seedi...
Embodiment 3
[0049] Embodiment 3: Using the method of the present invention to detect cell adhesion strength
[0050] 1. Prepare GFP-labeled mouse calvarial bone cell line MC 3T3-E1 at 4×10 4 piece / cm 2 Cells were seeded in 96-well plates at a density of 1.4×10 4 50 wells were inoculated, and 200 μl of medium was added to each well. Place in a 37°C cell culture incubator for 24 hours to grow to a confluence of 80%;
[0051] 2. Take the EDTA solution as the standard solution, dilute it twice and a half, and prepare a concentration gradient solution. The concentrations of each gradient solution are 50%, 25%, 12.5%, 6.25%, 3.13%, 1.57%, and 0.79% of the concentration of the standard solution;
[0052] 3. Drain the medium in 45 wells of the 96-well plate, and treat the cells with PBS and EDTA concentration gradient solution (0.79%-100% standard solution concentration), 100 μl per well, and make 5 replicates for each concentration. The last 5 wells with culture medium were used as the contr...
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