Basics of Primary Cell Culture

Primary cells are extracted and cultivated in vitro directly from living tissues. These cells do not multiply greatly, implying that they have had a limited number of passages and hence possess native properties of parent tissue. Primary cells and cell lines are not the same things because cell lines are developed from immortalized cultures of primary cells.

The concept of using primary cells in the clinical translational area is picking up steam in biomedical research. Primary cells have numerous applications in tissue engineering, 3D modelling, cancer research and cell therapies.

However, when it comes to dealing with primary cells in the lab, researchers confront a variety of challenges. This blog provides a few basics of primary cell culture to help alleviate these issues and make working with primary cells easier.

Cell Growth Requirement

Both suspension (cells that do not require a solid surface to develop) and adherent (cells that do require a solid surface to grow) cultures can support primary cell growth. Because the risks of contamination in primary cell cultures are higher, antibiotics must be added to the growing media.

Primary Cell culture handling abilities should be outstanding for a longer life span of the cells; also, adequate culture parameters such as pH, growth factors, concentration, growth media, temperature, presence of nutrients, and glucose are required. It is advised to use the aseptic approach.

Maintaining Cell Subculture

Cell maintenance starts immediately as they are isolated and adhered to the surface of the culture medium. Cell attachment takes roughly 12 hours from the time of seeding. It is time to subculture when the appropriate confluence percentage is reached and the cells are vigorously proliferating. The optimal time to start sub culturing primary cell cultures is when they are 80–90% confluent, as fully confluent cells may undergo differentiation and begin to grow more slowly following sub culturing.

The Confluence of Cell Culture

In adherent cultures, cellular confluence refers to the surface area of the culture vessel that is colonized by attached cells. For instance, 100% cellular confluence implies that cells cover the whole surface area of the vessel, whereas 50% confluence means that cells cover only half of the surface area. It is an important and critical parameter to monitor since different cells require distinct confluence endpoints or points at which the cells must be sub-cultured.

Cryopreservation

Cryopreservation is a technique that uses low temperatures to preserve living cells. This is usually done to extend the shelf life of primary cells. Cells must be cryopreserved appropriately in order to minimize cell damage and death throughout experimental activities. In the case of primary cells, DMSO is commonly applied in conjunction with Fetal Bovine Serum as a cryoprotectant. For more information on cryopreservation, you can read the following article https://kosheeka.com/best-tips-on-cell-culture-cryopreservation/.

If you are looking for primary cells for your research, then contact Kosheeka at info@kosheeka.com for further insights.