1. Primary cells are extracted directly from the source organ or whole blood.
Ethical donation of human organs and tissues allows for the cells to be investigated independently from the whole body. This provides important information on the cell’s function and behaviour and allows physiologically relevant scientific advances. The use of donated tissue also avoids the ethical issues associated with animal experiments.
2. The tissue can be mechanically or enzymatically disaggregated.
The method of disaggregation is dependent on the condition of tissue to be broken down. For example, some tissue types are tough, like bone or skin, and require a more mechanical manipulation, while softer tissues such as mammary tissue which can be digested solely by proteolytic enzymes.
3. Washing steps are important to ensure a sterile isolation of cells.
It is common to wash the tissue samples and disaggregated cells in antibiotic and antifungal solutions to prevent biological contamination, which can be detrimental to your cell culture. It is important to practice good aseptic technique and perform primary cell isolation separate from any other cell culture activities.
4. Knowing the growth conditions for your primary cells is essential.
Understanding the requirement for your primary cells will lengthen their life span and increase their reliability as a model for your research. Should they be grown in suspension or adherent to your culture vessel? Which culture vessel should they be grown in? What concentrations of growth factors and glucose do they require? What pH is optimal for their growth? These are some essential questions to consider before initiating a primary cell culture.
5. Primary cells have a natural capability and lifespan in culture.
Primary cells have a huge advantage over cell lines in that they represent their tissue of origin more closely and therefore provide more physiologically relevant results. It is important to remember that they are not cell lines, and in order to provide you with optimal results, they cannot be cultured over too many passages or they may become senescent. It’s important to understand their advantages and use these properly to improve your practise with primary cells.
6. Master Cell Banks (MCB) and Working Cell Banks (WCB) make the most of the cells’ potential.
Preparation of an MCB of your primary cells, perhaps from the initial tissue sample i.e. at passage 0, and a WCB to draw from for each experiment, allows the use of cells which have undergone the same handling and growth periods, and minimises the effect of this variability on your data. Careful planning of MCB and WCB production maximises the potential research that can be carried out using donor tissue.