Ways to Improve your Cell Culture Yields
Cell culture has advanced enormously since the time of its early development around 1900. Today it has been considered as one of the most enthralling branches of life science research. Almost 95% of the research papers published all over the world incorporate cell culture tools in their studies; which forces us to conclude that scientific advancements would have been almost impossible without cell culture techniques. Today, cell culture has been extensively employed to study different tissues, viral biology, the pathophysiology of different diseases, cancer development, aging processes, degenerative diseases, etc. Recently, scientists are relying upon the large-scale manufacturing processes of different primary cells to use them in different biopharmaceutical processes; another growing facet of biotechnology.
Despite being such an advanced science and being widely used, many of us might not be knowing some basic facts about the right cell culture practice. Scroll on to know more about the right cell culture practice; are you ready to learn something new?
· Workflow for the subculture of adherent cells
Different streams of biomedical research require the use of different primary cells, isolated directly from the tissues, such as gingival fibroblast cells, endothelial cells isolated from the umbilical cord, etc. Human cell cultures are essential tools in biomedical research, wherein different drug discoveries are being currently proposed using them, further avoiding the use of clinical studies. These cells allow rapid growth and proliferation of different types of cells for experimental analyses. Thus, they need to grow under optimized conditions, along with their specific growth medium. To achieve better consistency, all cells must be sub-cultured when the cells are reaching 80% confluency. During subculture, proper growth potential and seeding density of cells can be considered as major factors playing an important role.
· Why do I need to split my cells?
Once the primary cells are being isolated from mature tissues, they cannot be grown for an infinite period. The fully grown cells consume a lot of nutrients present in the optimized growth media, forcing them to produce a lot of toxic metabolites, eventually causing major cell death. It is also quite possible that researchers want more and more experiments, several times; for which, cell number matters a lot. Subculturing of cells cannot only increase its number but also maintains its metabolic activity. After initial seeding, the growth starts with the lag phase and move on to the log phase. This is the phase, wherein cellular proliferation takes place exponentially followed by a stationary phase where growth rate and death rates are equal.
· Why do I need to observe my cells under the microscope?
It is very important to examine a cell culture every day and essentially before following their subculture; to monitor their health as well as growth. Cells are first examined for fungal/bacterial contamination, unexpected transitions in their pH, general cellular morphology, etc. It has been advised that most of the cellular observation should be done with the help of a phase-contrast microscope because many cells are difficult to observe using a light microscope. Although mammalian cells are present of variable nature, however, three most general classifications are fibroblastic cells, epithelial-like cells, as well as lymphoblast cells. Additionally, certain cells have morphological characteristics, like neurons are easily identified due to their long dendritic extensions; cardiomyocytes are beating fibroblasts like cells.
· How can I subculture my cells?
One of the most commonly employed methods for subculture is using 0.25% trypsin with EDTA to detach cells from the plastic surface. EDTA acts as a calcium chelator that removes calcium from the cell culture, which further helps remove cadherin junctions that are involved in cellular interactions and cellular anchoring.