Comprehensive Summary of MKN-45 Cells

MKN-45 cells were established by Hojo H and originated from the liver metastasis of a poorly differentiated gastric adenocarcinoma in a 62-year-old female patient in Japan. These cells are widely used in research related to cancer detection and treatment.

In this issue of Cell Culture Academy, we have compiled information on the morphology and culture methods of MKN-45 cells to help you quickly get started with your experiments.

Cell Morphology

MKN-45 cells exhibit a semi-adherent, semi-suspension growth pattern. Some cells form loose aggregates, and the arrangement of cells is irregular. The edges of growing cells are also indistinct.

Microscopic images of MKN-45 cells, 100X

Culture Methods

MKN-45 cells exhibit a semi-adherent, semi-suspension growth pattern, requiring separate handling for the suspension and adherent cells.

Suspension Cells

Directly collect the culture medium into a centrifuge tube and centrifuge at 1200 rpm for 3 minutes.

Adherent Cells

Handle adherent cells as follows:

1.  Add approximately 2 ml of PBS to the culture flask, gently shake to rinse the cells, and aspirate the PBS (containing a small number of suspension cells) into the centrifuge tube used for suspension cells.

2. Add approximately 1 ml of 0.25% trypsin containing EDTA, and gently shake the culture flask to ensure all cells are covered.

3. Place the flask in the incubator for 3-5 minutes. Under a microscope, observe the cells; when they start to round up and detach from each other, stop the digestion process. This is indicated when the cells can slide off the upright flask wall. If cells are not rounded and do not slide off, extend digestion by 30-60 seconds. Avoid tapping the flask during this process.

4. Add 3 ml of serum-containing culture medium to stop the digestion. Pipette the cells up and down to detach them and to create a single-cell suspension.

5. Collect the cell suspension and centrifuge at 1200 rpm for 3 minutes. After centrifugation, discard the supernatant.

Techniques and Precautions

1. Since MKN-45 cells exhibit a semi-adherent, semi-suspension growth pattern, it is important to collect the floating cells during digestion and passaging to avoid cell loss.

2. If large cell aggregates are present in the suspension, collect the suspension into a centrifuge tube, centrifuge it, and then digest the aggregates with trypsin. This prevents the increase in cell clumps, which can affect cell growth.

Detailed Operating Steps:

1. Transfer all the culture medium from the culture flask into a sterile centrifuge tube. Centrifuge at 1200 rpm for 3 minutes to collect the cells.

2. Remove the supernatant and resuspend the cells in PBS. Collect the cells into a single centrifuge tube.

3. Centrifuge again, discard the supernatant, and resuspend the cells in approximately 1 ml of 0.25% trypsin. Mix gently by inverting the centrifuge tube; do not pipette. Place the tube in the incubator to digest the cells for 2-3 minutes, adjusting the time based on the specific cell characteristics.

4. After digestion, gently pipette the cell suspension to disperse cell clumps.

5. Add 5 ml of serum-containing culture medium to neutralize the trypsin and mix well. Centrifuge at 1200 rpm for 3 minutes.

6.  Collect the cell pellet, mix it with the portion of cells obtained from the adherent digestion, and then split and seed the cells into new culture flasks.