How to Perform Suspension Cell Culture

graduated conical flaskCells grown in the laboratory can generally be divided into two simple categories: adherent cells and suspension cells. The growth of adherent cells must have a support surface to which they can attach. Adherent cells are cells that can grow on a support surface only by means of attachment factors secreted by themselves or provided in the culture medium. The growth of suspension cells does not depend on the surface of the support, and it grows in suspension in the culture medium.

Suspension cells have good dispersibility. And they observed under the microscope are generally single growth of uniform size, round and bright cell shape, and some of them will aggregate and grow in clusters. Suspension cells grow rapidly with short doubling times. Suspension cells can directly perform some protoplast isolation, hybridization, and secondary metabolite production.

Suspension Cell Culture

The non-adherent suspension-grown cells can be detached from the surface of the culture vessel without the action of enzymes, so its subculture method is relatively simple. Direct passaging and centrifugation passaging are two methods commonly used to handle suspension cells in general laboratories.

When one observes that the cells in suspension have grown to 80% to 90%, the cell suspension can be seen turning yellow and the cells can be passaged. Materials required for the experiment are a culture vessel containing cells in suspension; a complete growth medium and preheated to 37°C; a 37°C incubator with humidified air with 5% carbon dioxide concentration.

Direct Passaging Cell Culture

  • If you observe basically no cell debris from the microscope, and the cell morphology is regular, it means that the cells are in good condition. You can choose the direct passaging method.
  • Remember to discard 1/2-1/3 of the cell suspension with a pipette.
  • Divide the medium from the original bottle proportionally into new culture bottles. This ratio depends on the experiment, but it should not be so thin that the cells cannot grow.
  • Then add fresh medium to continue culturing. The amount of medium supplemented should not be too high, as this will cause the cells to fail to grow due to low density.
  • And observe the cell density on the next day to consider whether to continue to replenish the liquid.

Centrifugation Passaging Cell Culture

  • However, when cell debris is observed and clearly numerous, and when the cell morphology is irregular, these indicate that the cell state is not good. In this case, centrifugation passaging is required.
  • First, transfer the cell suspension to a centrifuge tube. The supernatant was then discarded after centrifugation at 1000 rpm for five minutes.
  • Bounce the cell pellet gently, try not to use too much force or the cells will be damaged.
  • Resuspend cells in a fresh medium.
  • Finally, pipette an appropriate amount of cell suspension into a new culture flask and add an appropriate amount of fresh medium to continue the culture.

The above is about the culture of suspension cells. If you need to purchase cell culture-related equipment in cell culture-related experiments, you can cast a glance over Hawach’s cell culture consumables. Hawach cell culture consumables are available in a full range of cell culture dishes, cell culture plates, cell culture flasks, and Erlenmeyer flasks.

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