what is the difference between a primary culture and a cell line

Difference Between Primary Cell Culture and Cell Line

• Definition. ...
• Origin of Primary Cell Culture and Cell Line. ...
• Alterations in the Genetic Makeup. ...
• In Vivo Model. ...
• Lifespan of Primary Cell Culture and Cell Line. ...
• Maintenance. ...
• Donor Characteristics. ...
• Function. ...
• Importance. ...
• Conclusion. ...

Full Answer

What are primary cell lines?

Primary cell lines are obtained after dissociating tissues and organs but have a short life due to contact inhibition of the cell monolayer. These are used for inoculation of Orthomyxoviruses and echoviruses. The subculture of primary cell monolayers yields semi-continuous cell lines, which survive up to 50–100 population doublings before dying.

What is a primary cell culture?

Unlike 2D environments, a 3D cell culture allows cells in vitro to grow in all directions, similar to how they would grow in vivo. These three-dimensional cultures are usually grown in bioreactors, small capsules where the cells can grow into 3D cell colonies or spheroids.

What is primary cell?

Robustly generated from unlimited source material, these cells more closely resemble primary human hepatocytes, cells commonly taken from the liver of donors to produce a "liver-in-a-dish" for ...

What are primary cells?

Primary cells are an attractive cell source as the cells are terminally differentiated and already contain the intracellular machinery required to secrete the therapeutic protein. They are typically nonproliferative and genetically stable.

What is a cell line in cell culture?

Cell line is a general term that applies to a defined population of cells that can be maintained in culture for an extended period of time, retaining stability of certain phenotypes and functions. Cell lines are usually clonal, meaning that the entire population originated from a single common ancestor cell.

Why are cell lines better than primary cells?

Immortal cell lines are often used in research in place of primary cells. They offer several advantages, such as they are cost effective, easy to use, provide an unlimited supply of material and bypass ethical concerns associated with the use of animal and human tissue.

What are primary human cell lines?

Primary Cells – Cells isolated directly from human or animal tissue using enzymatic or mechanical methods. Once isolated, they are placed in an artificial environment in plastic or glass containers supported with specialized medium containing essential nutrients and growth factors to support proliferation.

What is primary culture in cell culture?

Primary Culture. Primary culture refers to the stage of the culture after the cells are isolated from the tissue and proliferated under the appropriate conditions until they occupy all of the available substrate (i.e., reach confluence).

What is the difference between primary cells and secondary cells?

Primary cells are the ones which cannot be recharged and have to be discarded after the expiration of the lifetime whereas, secondary cells need to be recharged when the charge gets over. Both the types of battery are used extensively in various appliances and these cells differ in size and material used in them.

How are cell lines generated from primary cultures?

If normal cells are kept in culture and allowed to divide and expand in number, either they become senescent, if they are normal cells, or mutations occur that allow the cells to continue to divide and be passaged. At that point a cell line has been generated.

What are cell lines in microbiology?

The term cell line refers to the propagation of culture after the first subculture. In other words, once the primary culture is sub-cultured, it becomes a cell line. A given cell line contains several cell lineages of either similar or distinct phenotypes.

Where do primary cell lines come from?

Human primary cell cultures can be initiated from healthy cells or cancer cells. They come from healthy donors, organ donation, surgical specimens, fetal tissues or post-mortem donors.

What are different types of cell lines?

Cells are classified in 4 different cell type categories based on overall morphology 1) Epithelial 2) Endothelial 3) Neuronal or 4) Fibroblast.

What is cell line example?

HeLa cells are an example of an immortal cell line. These are human epithelial cells obtained from fatal cervical carcinoma transformed by human papilloma virus 18 (HPV18). Indefinite cell lines are easy to manipulate and maintain. However these cell lines have a tendency to change over a period of time.

What is the difference between cell line and cell strain?

Cell line refers to a cell culture developed from a single cell and therefore, consist of cells with a uniform genetic make-up while cell strain refers to the cells derived from a primary culture or a single cell (clone) and possesses a specific feature such as a marker chromosome, antigen, or resistance to a virus.

Why is primary cell culture important?

Primary cell culture is important in immunology, vaccination, inflammation, etc. as it closely matches the in vivo function. Meanwhile, a cell line is important to study tumor cells when the cells of interest are not available as primary cell cultures. Hence, this is another difference between primary cell culture and cell line.

What are some examples of primary cell culture?

Some examples of primary cell culture are epithelial cells, endothelial cells, keratinocytes, melanocytes, fibroblasts, muscle cells, mesenchymal cells, hematopoietic cells, etc. Significantly, the cells in primary cell culture have the same karyotype as the cells in the parent tissue. Furthermore, the two main types of primary cell cultures based ...

What is the difference between a primary cell and a cell line?

Primary cell culture refers to the growing and maintaining of the selected cell type which has been excised from the normal parental tissue while a cell line refers to a cell culture developed from a single cell and therefore consisting of cells with a uniform genetic make-up. Thus, this is the main difference between primary cell culture and cell line.

What makes a cell immortal?

Generally, the cells of a cell line have undergone modifications such as random mutations or deliberate modifications such as the artificial expression of the telomerase gene. It makes the cells immortal with an unlimited lifespan in the culture.

Why do cells in primary cell culture have a limited lifespan?

However, the cells in primary cell culture have a limited lifespan due to the exhaustion of substrate and nutrients, gradually increase in the level of toxic metabolites, etc.

Why are cell lines important?

On the other hand, a cell line is a modified type of primary cells in order to be immortalized in the culture. Therefore, the cells in a cell line have an unlimited lifespan and become capable of subculturing. Generally, cell lines can be used to study tumor cells ...

What are the two types of cell cultures with different characteristics?

Primary cell culture and cell line are two types of cell cultures with different characteristic features.

What is the difference between a cell line and a primary cell culture?

Answer. Primary cell culture is the culture of cells directly isolated from parental tissue of interest; where as cell line is the culture of cells originated from a primary cell culture , which is generally used to expand cell population and prolong life span. These two processed differ in a few aspects.

How are cells obtained in primary cell culture?

Process of obtaining cells: In primary cell culture, cells are isolated from tissues, which usually go through phases of rinsing, dissection, mechanical or enzymatic disaggregation, and separation. In contrast, obtaining cells for a cell line is much more straightforward, which are directly transferred from the primary cell culture. If the primary cell is an adherent type, a detaching step is required.

Why do primary cells have a finite life span?

Life span: Primary cell cultures have finite life spans because the growth of cells exhausts substrate and nutrients, during which toxic metabolites are also accumulated, leading to the death of cells. However, cell lines have prolonged lifespan.

Can subcultures produce immortal cells?

However, cell lines have prolonged lifespan. Periodic sub-culturing can even produce immortal cells through transformation or genetic alteration of primary cells. Risk of contamination: Primary cell cultures are more difficult to take care of, which has a higher risk of contamination than the cell line.

What are the factors that are considered when performing primary cell culture?

When performing primary cell culture, researchers are acquiring an opportunity to study donors and not just cells. Several factors such as age , medical history, race, and sex can be considered when building an experimental model.

What are the two types of cells used in cell culture?

Cell culture procedures are conducted with two types of cells: Primary Cells – Cells isolated directly from human or animal tissue using enzymatic or mechanical methods.

What is a cell line?

Cell Lines – Cells that have been continually passaged over a long period of time and have acquired homogenous genotypic and phenotypic characteristics. Cell lines can be finite or continuous. An immortalized or continuous cell line has acquired the ability to proliferate indefinitely, either through genetic mutations or artificial modifications. A finite cell line has been sub-cultured for 20-80 passages after which they senesce. Cell lines are preferably used for convenience as they are easy to handle and widely published. However, they are less preferred as a biologically relevant option, since they have lost the true characteristics of the original tissue from which they were isolated. Serial passaging is known to cause genotypic and phenotypic variation in cell lines. Variation can often be so far from that of the original tissue to where they do not mimic the in vivo environment very closely. Cells that do not represent the original tissue could result in false negative or false positive findings.

Where are adherent cells usually derived from?

The adherent cells are usually derived from tissues of organs. Suspension cells do not require attachment for growth and are said to be anchorage-independent cells. Most suspension cells are isolated from blood system.

What type of cells are used for ADMET?

Some tissue-derived suspension cells are available such as hepatocytes or intestinal cells which are suitable for ADMET applications. Although primary cells usually have a limited lifespan, they offer a huge number of advantages compared to cell lines.

How long have cell lines been in culture?

Most cell lines have been in culture for decades and are well adapted to the two-dimensional culture environment, and as a result, often differ genetically and phenotypically from their tissue origin and show altered morphology [3,4].

Why is cell culture important?

Cell culture studies provide a valuable complement to in vivo experiments, allowing for a more controlled manipulation of cellular functions and processes. For decades, cell lines have played a critical role in scientific advancements, yet researchers have become increasingly cautious when interpreting data generated from cell lines only. Factors such as misidentified and contaminated cell lines have spurred renewed interest in primary cells [1,2]. Many researchers have chosen to work with cells lines as they are generally highly proliferative, and easier to culture and transfect. Most cell lines have been in culture for decades and are well adapted to the two-dimensional culture environment, and as a result, often differ genetically and phenotypically from their tissue origin and show altered morphology [3,4]. In contrast to cell lines, primary cells which are isolated directly from tissues, have a finite lifespan and limited expansion capacity. On the positive side, primary cells have normal cell morphology and maintain many of the important markers and functions seen in vivo [3,4]. Endothelial cell lines, for example, lack various functional markers, while primary endothelial cells retain these critical features.

What is the role of serum in cell culture?

Traditional cell culture media has relied on serum to provide the growth factors, hormones, lipids and other undefined components to support cellular growth. For primary cells, however, high serum levels can lead to differentiation or promote growth of contaminating cells like fibroblasts.

Is it harder to work with primary cells?

Although primary cells may be more difficult to work with, the data obtained from using primary cells is more relevant and reflective of the in vivo environment.

Do primary cells have a limited lifespan?

In contrast to cell lines, primary cells which are isolated directly from tissues, have a finite lifespan and limited expansion capacity. On the positive side, primary cells have normal cell morphology and maintain many of the important markers and functions seen in vivo [3,4].

Why do you need to purify cells for a primary culture?

Also, primary cultures derived from tissue explants are a mix of different cells at different stages, so if you want to initiate more homogeneous cultures, you might need to purify specific cell types. Because primary cells are more sensitive than cell lines, they often require additional nutrients and growth factors.

Where do primary cells come from?

They come from healthy donors, organ donation, surgical specimens, fetal tissues or post-mortem donors. When planning your experiments, keep in mind that the source of human primary cells is limited.

What are the causes of cell culture contamination?

Contamination threatens cell culture studies. Not only bacteria, fungi, yeast and mycoplasma are the culprits, endotoxins also cause problems. They affect cellular growth and function in vitro as well as in vivo ( Ryan et al, 2018 ). Endotoxins are lipopolysaccharides derived from the outer membrane of most gram-negative bacteria, which are stable even at high temperatures. Endotoxins often contaminate labware, as they show high affinity for hydrophobic materials such as plastic. Sources of contamination include water used for washing, media and sera, additives, laboratory plastics and glassware. You can detect endotoxins with the limulus amebocyte lysate (LAL) assay, as LAL forms quantifiable gel-clots in their presence.

Why do we need to optimize cell culture?

Because different types of cells need different media to grow and survive, always optimize culture conditions for each cell type. Cell lines need high levels of serum. However, serum is not standardized and various lots can display high variability of the many substances contained. This could greatly influence the results of your research and even make them inconsistent. In the case of human primary cells, remember that high levels of antibiotics decrease their viability and growth. As cells reach the different stages of culture, you need to characterize them and identify any morphological and functional changes. This is relevant for quality control. Of course, good cell culture practice demands that you document all the information necessary to track the materials and methods. In this way, you ensure that your model is transparent and reproducible ( Pamies et al., 2018 ).

How many cell lines are misidentified?

studies estimate that 18-36% of all cell lines are misidentified. Working with primary cells rests upon decades of research using cell lines. Since the early 20th century, cell lines have provided scientists with great insights into the biological processes of human cells. Immortal cell lines have become a powerful tool for countless applications ...

Why use immortal cells?

Then use human primary cells to replicate key findings. This increases translation and makes your results more relevant to human physiology.

Why do scientists use cell lines?

In research and drug development, scientists routinely use cell lines as a model for healthy or diseased tissue. Cell lines offer an easy, inexpensive and stable platform. However, they often do not fully represent what is occurring in vivo. If you want to model the complex physiological behavior, human primary cells might be the better choice.

What is the difference between primary and secondary cell culture?

The main difference between primary and secondary cell culture is that the primary cell culture contains the cells directly obtained from host tissue, whereas the secondary cell culture contains sub-cultured cells from primary cell culture. Furthermore, cells are dissociated from the host tissue either through mechanical or enzymatical digestion.

What are the two types of primary cell cultures?

Furthermore, there can be two types of primary cell cultures according to the type of cell growth. They are adherent cultures and suspension cultures . Generally, adherent cell cultures are anchorage-dependent and require an attachment for the growth while suspension cultures are anchorage-independent and grow in the medium. However, primary cell cultures have a limited lifespan. Usually, cells undergo programmed cell death due to contact inhibition, the requirement of survival factors. Typically, they build up toxins with the population growth while cells require growth factors for their survival, respectively. Therefore, primary cell cultures can only be maintained only for a few generations or passages in culture.

What is primary and secondary cell culture?

Primary and secondary cell culture are two types of cell cultures which contain living cells grown in a medium in vitro under controlled conditions. Both allow the growth of cells and the maintenance of regular cell functions to a greater extent.

Why do primary cell cultures have a limited lifespan?

However, primary cell cultures have a limited lifespan. Usually, cells undergo programmed cell death due to contact inhibition, the requirement of survival factors. Typically, they build up toxins with the population growth while cells require growth factors for their survival, respectively.

How are cells in primary and secondary cell cultures adapted to the culture conditions?

Cells in primary cell cultures have the same biological response as the cells in the host tissue while cells in secondary cell culture are adapted to the culture conditions by altering their biology.

Why do cells have an indefinite lifespan?

The indefinite lifespan is due to the acquisition of immortality through mutations or viral transformation. In addition to these, due to the providing of survival factors continuously, cells can acquire an indefinite lifespan. Therefore, further proliferation of cells is stimulated in secondary cell cultures.

What is sub-culturing in biology?

Typically, sub-culturing is the transferring of cells in a primary cell culture into a new vessel with a fresh medium. However, cells may spontaneously acquire mutations when it goes through several sub-culturing steps.

What Is A Primary Cell Culture?

What Is A Cell Line?

What Are The Similarities Between Primary Cell Culture and Cell Line?

Summary – Primary Cell Culture vs Cell Line