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Since the discovery of cell culture in 1907 it has rapidly become one of the most frequent and important techniques used by cell biologists and as more modern technology has became available, a greater understanding of the method was gained to further enhance the results of cell culture. With the developments of two-dimensional cell culture to three-dimensional cell culture a plethora of breakthrough discoveries have occurred within disease, stem cells and drug activity.
Cell culture refers to the transfer of cells from a specimen to a favourable, nutrient rich environment to optimise growth in which they can be grown in two-dimensions (2-D) or three-dimensions (3-D) (1). Traditionally, cells were grown in 2-D on a plastic plate resulting in a monolayer that caused the cells to become flat in shape, modifying gene and protein expression (2). In recent years the cell biology world has received a breakthrough in the form of 3-D cell culture which allows the cells of a specimen to interact with the environment to a greater degree than 2-D cell culture (1). This allows cells to precisely replicate cells in vivo state.
The breakthrough of 3-D cell culture has created opportunities for new discoveries to be made in regards to cancer, drug testing and neurodegenerative diseases.
3-D culture systems have enabled the replication of in vivo growth environment of cancer in vitro. Interestingly, cancer cells now being grown within the extracellular matrix of a culture, accurately represent the 3-D cell environment and can allow researchers to observe the disorganised structure of cancer cells (3). Cultures are often begun by dissociation of the cancer tissue of the specimen and placed on the culture surrounded by non-cancerous components to accurately represent the tumour microenvironment so as the cultured cells retain the essential features of the primary cancer (4).This is particularly useful in studying the characteristics of tumours and tumour spheroids which have clinical importance in their chemotherapy resistance that may be associated with malignancy in some cancers (5). The ability to observe the nature of cancerous cells and how they react to environmental stimuli will allow for the development of more effective pharmaceuticals to combat this prevalent disease (6).
Anticancer therapies are in huge medical demand due to the high level of diagnosis of cancers occurring worldwide. However, the amount of drugs available that have shown positive results are very few in number with chemotherapy and radiotherapy still being the greatest chance a patient has at surviving. Cell culture can be used to detect the effectiveness of anticancer therapies. The effectiveness of anticancer drugs was previously determined by 2-D cell culture which has now been recognised to have given false positive results in drug activity as the tumour microenvironment was not accurately stimulated (7). This has lead to 3-D cell culture to be used. In the study of the effect of anticancer drugs on breast cancer evaluating 2-D and 3-D cultures, it demonstrated that those grown in 3-D formed dense multicellular spheroids which was found to be associated with resistance to chemotherapy drugs. Those grown in 2-D over-estimated the drugs efficiency and were then disregarded during clinical development when the drug began to give negative results (7).
The huge developments in cell culture over the recent decades have enabled technologies to produce microphysiological systems or simply tissue chips that imitate the function of human organs and that react to physiological inputs and outputs to maintain homeostasis. Tissues are grown on chips of clear plastic under 3-D culture to allow observation of cell processes and reaction to variable stimuli through a microscope (9). This technique will prove to be useful in drug development and drug screening processes as it will eliminate the ethical issue for animal testing as it may no longer be required due to the effects of the drug being observable under a microscope (10). Furthermore, when human clinical trials begin the drug will have had effective screening and modification to inhibit disastrous consequence (9).
Stem cells are constantly under the spotlight in the scientific world due to their ability to differentiate into any cell type and their self-renewal properties. These cells once hard to retrieve without invasive techniques such as through the bone marrow, have now become increasingly easy to access through the discovery that adult somatic cells can be used to obtain induced pluripotent stem cells (iPSCs) after reprogramming, through skin biopsies (11). Following on from tissue chips, brain organoids can be developed through iPSCs to study neurodegenerative diseases. Due to the nature of neurodegenerative diseases often occurring on an aged brain, the cell culture of the organoid needs to represent this state. However, studies have shown that creating an aged brain in culture does not capture every aspect of its characteristics, it can however demonstrate late progression in the disease with earlier progression not being accurately modelled (12). The visualisation of neurodegenerative diseases have provided researchers with information to develop personalised medicine and accurate drug screening for patients with Alzheimer’s, Parkinson’s, Huntington’s disease and many more (13).
Brain organoids created from iPSCs from donors have already led to breakthrough discoveries in learning disabilities and newly discovered target genes in autism spectrum disorders (14) so it is hoped with continual study of iPSCs it will give rise to regenerative medicine that could revert a diseased brain back into its healthy state.
Cell culture is an essential technique for researchers to utilise to make new discoveries within cell biology, biochemistry and biomedical science. Continual research within cell culture has the potential to change the future of clinical medicine by providing a greater understanding of terminal diseases and the development of new drug therapies, more effective at targeting the issue. Regenerative medicine is no longer an idea; through cell culture it is becoming a reality in regards to neurodegenerative diseases and spinal cord injuries by the modelling of organs in microphysiological systems in vitro. Without cell culture we would not be able to entirely understand the physiology and biochemistry of cells creating a barrier in knowledge essential in understanding organisms from tiny bacterial cells to the more complex human body.
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