You are currently viewing Basics Of the Primary Cell Culture That Are Valuable for Your Research

Basics Of the Primary Cell Culture That Are Valuable for Your Research

You may be surprised if we say “Cells present in your body have the potential to structure the research”.

Yeah, you hear us right 

The development of cell culture techniques significantly transformed modern medicine. Research with the use of cell lines has become an integral part of the development of medicine and therapeutic establishment. Over the period, the methods of cell cultures have also evolved 360 degrees. The raw materials that were used earlier have been replaced with advanced techniques and tools. 

Over the period, the methods of tissue culture have transformed 360 degrees. The raw materials that were used earlier have been replaced with tools, designed by modern technologies. For the very first time, cell culture observation was done in the 18th century by Willhelm Roux, who was able to maintain chicken embryos alive in an in-vitro set up in saline for a few days. However, the exact understanding related to primary cell culture developed at the beginning of the 20th century, when frog nerve cells were successfully cultured by the hanging drop method by Ross Harrison. Post this experiment, other successive discoveries like aseptic techniques, biosafety cabinets, centrifuge, etc. perfected the art of cell culture. 

Today, due to regulatory obligations associated with animal research, researchers are diverting their attention towards the primary cells, which can mimic the in vivo environment to a great extent. Some of the important advantages of using primary cells are:

When comparing the properties of primary cells and continuous cell lines:

  • Biological Relevance: Primary cells exhibit higher biological relevance as they closely resemble the tissue of origin, whereas continuous cell lines may deviate from the in vivo state due to genetic changes.When comparing the properties of primary cells and continuous cell lines:
  • Genetic Integrity: Primary cells retain the genetic makeup of the in vivo tissue, whereas continuous cell lines are subject to genetic drift and changes.
  • Native Phenotype: Primary cells maintain their origin phenotype, markers, and cellular pathways, even when cultured for subsequent passages. Thus, the natural state of cells, morphology, physiology, and communication channels are always maintained, even outside the body. 
  • Biological Diversity: Different donors or species can be sourced to isolate primary cells and expand them further, allowing scientists to explore biological variations and responses in a more diverse setup and representative manner. 
  • Early Disease Models: Most of the primary cells can serve as the early disease models, as they can be directly obtained from the patients; helping researchers to understand pathophysiological mechanisms in a deeper sense. 
  • Ease-of-Use: Primary cells maintain their functional integrity and physiological relevance even outside the body, and hence are less likely to entertain genetic as well as phenotypic variations, as compared to their immortalized cellular counterparts, allowing for a more direct correlation between in vivo and in vitro. 
  • Translational Research: Findings from primary cell studies are often more translatable to clinical settings, contributing to the advancements of translational research. 
  • Time & Expense: Primary cells typically can be advantageous to finalize dosing and route of administration in animals; thus, minimizing the number of animals required for preclinical analysis, hence are proven to be cost-effective alternatives to established cell lines. 

In summary, the choice between primary cells and continuous cell lines depends on the specific research objectives and the balance between biological relevance and practical considerations such as ease of use and cost.

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Primary Cell Cultures Can Be Categorized Into Two Groups Adherent And Suspension Cell Cultures Based On The Type Of Cells Present In The Culture:

Anchorage-Dependent/Adherent cells:

These cells rely on a stable, biologically inert surface for adherence and growth. They require a solid and non-toxic surface for optimal growth, as they are challenging to cultivate as cell suspensions. Typically, these cells are obtained from tissues of organs where cells are naturally immobilized within the connective tissue. Examples of adherent cells include kidney cells and mouse fibroblast STO cells.

Anchorage-Independent/Suspension cells:

These cells can efficiently grow as cell suspensions and do not necessitate a solid surface for attachment. They can be cultivated continuously in liquid media to obtain fresh subcultures. The ability of these cells to grow in suspension depends on their source, as cells naturally existing in suspension within the body are well-suited for suspension culture. Examples of suspension cells include blood cells, which are vascular and remain suspended in the plasma.

Major Differences Between Primary Cell Culture And Secondary Cell Culture

Primary cell culture and secondary cell culture represent distinct stages in cell culture processes, each with unique characteristics and applications.

PropertiesPrimary cell culture Secondary cell culture 
Definition In primary cell culture cells are directly isolated from living tissue and cultivated. Secondary cell culture, often referred to as a cell line, involves the continued propagation of cells derived from a primary culture.
Source They obtained directly from tissues or organsThey are derived from primary cultures through a process of immortalization, which allows the cells to proliferate indefinitely.
Nature They closely resemble the in vivo state and retain the physiological properties of the tissue of origin.They may undergo genetic changes or modifications during the immortalization process, leading to differences from primary cells and potentially deviating from the in vivo state.
Lifespan They limited lifespan and finite proliferation capacity. They eventually undergo senescence and cease to divide after a certain number of passages.They have an unlimited lifespan and can be propagated indefinitely under appropriate culture conditions.
Applications They are valuable for studying normal physiology, biochemical processes, and responses to stimuli. They are particularly useful for modeling tissue-specific functions, drug screening, and toxicity testingCell lines are widely used in research and biotechnology for their ease of use, reproducibility, and scalability. They are particularly valuable for large-scale experiments, high-throughput screening, and the production of biological products.

 

Animal Cell Culture Procedure 

Animal cell culture procedure’s initial step starts with cell harvesting in which cells are isolated from the tissue via appropriate enzymes under aseptic conditions. Afterward following steps are involved:  

Cell Isolation and Cultivation:

  • Transfer isolated cells to a culture dish containing suitable growth media.Animal cell culture procedure’s initial step starts with cell harvesting in which cells are isolated from the tissue via appropriate enzymes under aseptic conditions. Afterward following steps are involved:  
  • Incubate the culture dish in a controlled environment for cell propagation.
  • Regularly monitor cell morphology and confluency.

Subculturing (Passaging):

  • Upon reaching 70-80% confluency, detach cells from the culture dish.
  • Passage cells by transferring a portion to fresh culture dishes with replenished media.
  • Maintain optimal cell density for sustained growth.

Preparation of Complete Growth Media:

  • Formulate growth media with essential nutrients, serum, and growth factors.
  • Incorporate antibiotics to prevent contamination.

Culture Vessels and Surface Treatment:

  • Utilize sterile plastic culture dishes or flasks for cell culture.
  • Pre-treat surfaces with extracellular matrix proteins like collagen or fibronectin to enhance cell attachment.

Cryopreservation and Contamination Control and Biosafety Measures are the final steps of the animal cell culture procedure which ensures the viability and reproducibility of the animal cell culture

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Ethical And Legal Considerations Before Proceeding To The Primary Cells

Human specimens are very valuable tools used largely in translational medicine, for studies related to health issues. Each tissue represents unique genotypical and phenotypical markers from the source, and are obtained during surgery, post-mortem, biopsy, etc. Most of these biospecimen comprise of DNA, cells, proteins, exosomes, etc. and are collected for the isolation and culture of primary cells, representing these biospecimen. However, there are numerous ethical concerns, while handling biospecimen for primary cell isolation; related to bioethics, tissue collection, transportation and storage. The approach towards the biological research is highly individualistic, further necessitates the need for respecting and considering donor’s morales under advisement.  As per the clinical trial act and legislation related to the use of human biospecimen for laboratory research, biological samples can be obtained only with the written, morale consent from the donor. The written informed consent should include a clause mentioning the right to withdraw the approval and also ability to alter the scope of the agreement. 

Storage And Disposal Of Human Biological Material

One should also note that the biospecimens of human origin should always be treated as the biohazard; and are either to be incinerated to disinfected before discard. As mentioned above, the collection of specimens is usually done in an appropriate buffer containing antibiotics, in order to preserve the physiological integrity of the sample. One should also note that the biospecimens of human origin should always be treated as the biohazard; and are either to be incinerated to disinfected before discard. 

In summary, while primary cells offer several advantages such as ethical compliance, relevance to in vivo signaling, and cost-effectiveness, they also come with limitations including time requirements, inter-donor variability, lack of in vivo environment, and associated costs and feasibility challenges. These factors should be carefully considered when choosing primary cell cultures for research purposes.

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