15 things about cell culture you might not know (2023)

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15 things about cell culture you might not know (1)

Scientific progress would be inconceivable without cell culture. This research tool has become irreplaceable in life sciences, and it has greatly evolved since the beginnings of cell culture history in the early 20th century. Biomedical research today increasingly relies on new approaches including 3D cell culture, primary cell culture, tissue engineering and regenerative therapies. Even if you deal with cell culture every day, chances are we’ve found a few facts about cell culture you didn’t know. Are you ready to learn something new?

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#1: Plastic can contaminate cell cultures

Apart from “known” contaminants such as viruses, bacteria, mycoplasma and endotoxins, the plasticizers eluted from plastic instruments, or a range of substances present in water, can also contaminate cells in culture and affect their behavior (Yao and Asayama, 2017).

#2: Frog nerve fibers were the first successfully cultured cells

In 1907, the American zoologist Ross Granville Harrison was the first researcher to successfully grow animal cells outside the body. He adapted the “hanging drop” method from bacteriology for use with tissue culture and managed to culture frog neuroblasts in a lymph medium (Abercrombie, 1961).

#3: More than 32,000 papers rely on cell lines with no known original stock

The use of misidentified cell lines is a very relevant problem in life sciences. Studies estimate that more than 32,000 papers report on results obtained with the wrong cells. Not only that, but up to half a million other papers have cited these reports, making the “contamination“ of literature a very serious issue (Horbach and Halffman, 2017).

#4: More than half of researchers fail to reproduce their own experiments

A Nature survey of more than 1,500 researchers showed that 70 percent of them were unable to reproduce another scientist’s experiments – and half of them failed even to reproduce their own experiments.

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#5: The first “immortal” cell line HeLa was established without the donor’s consent

15 things about cell culture you might not know (2)

Cells derived from a tumor biopsy of Henrietta Lacks, an African-American woman diagnosed with cervical cancer, were used to establish the first human cell line in 1951. However, tissue samples were taken without her permission or knowledge and her family was not made aware of the line’s existence until 1975. This issue raised many concerns about privacy and patients’ rights in biomedical research. Only in 2013, more than 60 years after her death, the National Institutes of Health and Lacks’ descendants reached agreement on the use of her cells and genetic information. This arrangement established a precedent in cell culture history and research ethics.

#6: Almost 25 percent of all cell line contaminations are caused by HeLa cells

In 1967, American scientist Stanley Michael Gartler examined 20 individual human cell lines he had collected for one of his projects at the University of Washington. He analyzed the glucose-6-phosphate de-hydrogenase (G6PD) and phosphoglucomutase (PGM) polymorphisms via electrophoresis and found out that all cell lines showed the same phenotypes and were therefore genetically identical. Moreover, the detected G6PD allele was found only in people of African descent, even though the original cell lines were derived from donors of Caucasian origin. This observation prompted Gartler to conclude that all cells were HeLa cells that had contaminated and grown over the other cells (Gartler, 1968). This realization caused a huge scientific dilemma and placed a large question mark over the cell culture-based findings of the previous 20 years of cell culture history. Still today, 24 percent of all contaminations among human cell lines are caused by HeLa cells (Lin et al., 2019).

#7: Cell culture history: The first modern medium was created more than 60 years ago

Medium 199 was developed by J.F. Morgan in 1950 and was one of the first synthetic media used to grow mammalian cells in culture (Morgan et al., 1950). The idea of designing a chemically defined medium without animal components made Medium 199 an ideal medium for vaccine production. It also allowed large-scale manufacturing of vaccines for the polio vaccination campaign in 1955. Nine years after Morgan, scientist Harry Eagle developed the minimum essential medium (MEM), which contained a mixture of glucose, salts, amino acids, and vitamins (Eagle, 1959).

#8: Components of culture medium can interact with each other and influence cultured cells

When you are trying to optimize your culture medium, consider this: Individual components of the medium do not act alone. Components can interact, and their effects on cells are not always predictable. This is particularly important when replacing animal sera in culture media. You might need to use mathematical algorithms to optimize the combination of multiple compounds and to establish the best conditions for cellular growth (Yao and Asayama, 2017; Kim and Audet, 2019).

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#9: “Mini-brains” can be grown from pluripotent stem cells

15 things about cell culture you might not know (3)

In a 2013 groundbreaking paper published in Nature, Dr. Madeleine Lancaster and her team developed a protocol to generate brain organoids, or “mini-brains” in vitro, starting from pluripotent human stem cells and using a 3D support matrix and a spinning bioreactor. These organoids contain different types of nerve cells and are structured much like mammalian brains. Furthermore, they show features of early human brain development (Lancaster et al., 2013). The same research team successfully created brain tumor models by introducing oncogenic mutations into the brain organoids (Bian et al., 2018).

#10: The year 2003 marks a milestone in bioprinting

Thomas Boland, a bioengineer at the University of Texas, El Paso, was working with an inkjet printer when he noticed that ink droplets were about the same size as human cells. This observation prompted him to fill an ink cartridge with living bovine cells, nutrients and other bio-compatible substances to create “bioink” that was capable of printing living tissues (Patel, 2016).

#11: Israeli researchers succeed in creating a 3D-printed human heart from fat tissue

15 things about cell culture you might not know (4)

In April 2019, Tal Dvir, a researcher at Tel Aviv University’s School of Molecular Cell Biology and Biotechnology and his team published a paper on the success of a new protocol for 3D-printing thick, vascularized, and perfusable hearts from a biopsy of human omental tissue. These hearts do not beat and are only the size of a rabbit’s heart, but they completely match the immunological, cellular, biochemical, and anatomical properties of the patients. These results represent a big step forward on the path to finding new treatments for cardiac diseases (Noor et al., 2019).

#12: Engineers and cancer researchers team up to develop a new 3D scaffold

An interdisciplinary team of researchers at the University of Michigan has created a network of fibronectin fibers interlacing across a grid of microscale cubicles. This structure could provide a natural environment for growing cancer cells from individual patients and for testing cancer drugs, two prerequisites for personalized therapeutic approaches (Jordahl et al., 2019).

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#13: Supercooling can increase preservation time of human organs

A recent article in Nature Biotechnology reports on a new method for supercooling and storing human livers at -4ºC without ice formation. This approach could extend the ex vivo life of organs from 12 hours currently to 27 hours, which would expand access to liver transplantation (de Vries et al., 2019).

#14: Artificial intelligence just might interpret your next large data set in cell biology

Major advances in artificial intelligence are turning deep learning into a research partner for biomedicine and cell biology. Recently, deep learning has successfully contributed to applications such as fluorescent staining prediction, cell type differentiation, bacterial resistance, and super-resolution microscopy (Waisman et al., 2019).

#15: The 3D cell culture market could reach nearly USD four billion in 2021

According to BBC Research, the global 3D cell culture market was valued at USD one billion in 2016 and should almost quadruple in 2021, thus showing an annual growth rate of around 30 percent. Increased investments in R&D, rapid advances in technologies, as well as an improved access to consumer healthcare, all drive market growth.


What is cell culture short answer? ›

Cell culture is the process by which cells are grown under controlled conditions, generally outside their natural environment. After the cells of interest have been isolated from living tissue, they can subsequently be maintained under carefully controlled conditions.

What are the 3 types of cell culture? ›

Cells cultured in the lab can be classified into three different types: primary cells, transformed cells, and self-renewing cells.

What is the importance of cell culture? ›

Cell culture is one of the major tools used in cellular and molecular biology, providing excellent model systems for studying the normal physiology and biochemistry of cells (e.g., metabolic studies, aging), the effects of drugs and toxic compounds on the cells, and mutagenesis and carcinogenesis.

What can go wrong in cell culture? ›

Generally, the most common issues include abnormal growth patterns or spotty, uneven, or inconsistent cell attachment. Other problems include slow or sudden changes in growth rate or unexplainable outcomes. Such issues tend to be associated with culture technique, incubation, and media.

What is cell culture made of? ›

Cell culture media generally comprise an appropriate source of energy and compounds which regulate the cell cycle. A typical culture medium is composed of a complement of amino acids, vitamins, inorganic salts, glucose, and serum as a source of growth factors, hormones, and attachment factors.

How long is cell culture? ›

As a general guide, from a confluent flask of cells: 1:2 split should be 70-80% confluent and ready for an experiment in 1 to 2 days. 1:5 split should be 70-80% confluent and ready for an experiment in 2 to 4 days. 1:10 split should be 70-80% confluent and ready for sub-culturing or plating in 4 to 6 days.

Who invented cell culture? ›

The American embryologist Ross Granville Harrison (1870–1959) developed the first techniques of cell culture in vitro in the first decade of the twentieth century [52–56].

What are the stages of cell culture? ›

There are four phases: lag, log, stationary and decline (29). During the lag phase, the number of cells is not increasing and the cells are simply acquiring enough resources to prepare for proliferation (29).

How many types of cell cultures are there? ›

There are three major types of cell culture, which include: Primary cell culture. Secondary cell culture, and. Cell line.

How does cell culturing work? ›

Cell culture is the growth of cells from an animal or plant in an artificial, controlled environment. Cells are removed either from the organism directly and disaggregated before cultivation or from a cell line or cell strain that has previously been established.

What equipment is used in cell culture? ›

What are the Basic equipments required for animal cell culture?
Essential equipmentsBeneficial equipmentsUseful additional equipments
IncubatorLaminar flow hoodLow-temperature freezer
MicroscopeCell counterGlassware washing machine
SterilizerVacuum PumpColony counter
Washing up instrumentCO2 incubatorClosed-circuit machine
6 more rows
Dec 16, 2020

What was the biggest problem facing cell culture? ›

But the biggest problem facing cell culture was contamination. Bacteria and a host of other microorganisms could find their way into cultures—from people's unwashed hands, their breath, and dust particles floating through the air—and destroy them.

What is one of the most common problems faced by researchers who use cell culture? ›

Contamination is a common problem in cell culture, and contaminants include mycoplasma, yeast, bacteria, and chemicals.

Why is cell culture contaminated? ›

Contamination may arise from the operator and the laboratory environment, from other cells used in the laboratory, and from reagents. Some infections may present a risk to laboratory workers: containment and aseptic technique are the key defence against such risks.

How do you maintain cell culture? ›

Cultured cells require a supply of nutrients for growth. Mammalian cell culture media must maintain physiological pH, in addition to providing balanced salts, carbohydrates, amino acids, vitamins, fatty acids and lipids, proteins and peptides, trace elements, and growth factors.

What is cell culture technology? ›

Cell culture involves a complex of processes of cell isolation from their natural environment (in vivo) and subsequent growth in a controlled environmental artificial condition (in vitro).

How do cells survive? ›

To survive, every cell must have a constant supply of vital substances such as sugar, minerals, and oxygen, and dispose of waste products, all carried back and forth by the blood cells. Without these substances, cells would die in a very short period of time.

Why is cell culture media yellow? ›

Cell cultures normally become acidic due to an increase in cell numbers and cellular respiration, resulting in a yellowing in color of media formulations containing phenol red.

What is cell culture and its types? ›

Types of cell cultures Primary culture :- refers to the stage (primary step) 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 At this stage, the cells have to be subcultured by transferring them t o a new ...

What is cell culture media used for? ›

Cell culture media and supplements are critical for supporting the maintenance and growth of cells in vitro, such as by maintaining extracellular pH. Essential components of cell culture media include carbohydrates, amino acids, vitamins, minerals, and a pH buffer system.

How can you tell if a cell is alive? ›

The most common way to identify dead cells is using a cell-impermeant DNA binding dye, such as propidium iodide or a dye from the STYOX series. A healthy living cell has an intact cell membrane and will act as a barrier to the dye so it cannot enter the cell.

How long can cells survive without CO2? ›

Mammalian cells in bicarbonate-based media require 5% CO2 to maintain physiological pH. Without a CO2 supply, cells are adversely affected within five minutes.

How do you collect cells? ›

Collect the cells by centrifugation at 300 x g for 7 minutes at 4°C. Aspirate the PBS. Lyse the cells by pipetting Complete Cell Extraction Buffer into each tube. We recommend using 1 mL of Complete Cell Extraction Buffer per 108 cells.

What is single cell culture? ›

Single-cell culture is a method of growing isolated single-cell routinely performed to obtain single-cell-derived cell clones for both basic research and therapeutically applications.

Where do cell lines come from? ›

Introduction. Cell lines are cultures of animal cells that can be propagated repeatedly and sometimes indefinitely. They arise from primary cell cultures. Primary cultures are initiated directly from the cells, tissues, or organs of animals and are typically used in experiments within a few days.

What are the most common cell lines? ›

HEK293, or human embryonic kidney-derived epithelial cells, are arguably one of the most widely used cell lines in cell biology research. HEK293 is a rapidly dividing, robust line cell with a good reputation for post-translational modification of its heterologously expressed proteins.

How do we produce new cells? ›

New cells are created from a process called cell division. The new cells are produced when a cell, called the mother cell, divides into new cells called daughter cells. When two daughter cells have the same number of chromosomes as the original cell, the process is called mitosis.

What are the two types of cell lines? ›

Finite and Continuous Cell Lines.

What are the characteristics of cell culture? ›

Characteristics of Cultured Cells:
  • Cells which do not normally proliferate in vivo can be grown and proliferated in cultures. ...
  • Cell to cell interactions in the cultured cells are very much low.
  • The three dimensional architecture of the in vivo cells is not found in cultured cells.

How long can cells survive without media? ›

It depends on cell type. However you can go upto 36-48 hours in general.

What is called cell culture? ›

Listen to pronunciation. (sel KUL-cher) The growth of microorganisms such as bacteria and yeast, or human, plant, or animal cells in the laboratory. Cell cultures may be used to diagnose infections, to test new drugs, and in research.

How do we define culture? ›

Culture can be defined as all the ways of life including arts, beliefs and institutions of a population that are passed down from generation to generation. Culture has been called "the way of life for an entire society." As such, it includes codes of manners, dress, language, religion, rituals, art.

What is culture in biology simple words? ›

culture. [ kŭl′chər ] n. The growing of microorganisms, tissue cells, or other living matter in a specially prepared nutrient medium. Such a growth or colony, as of bacteria.

What is plant cell culture? ›

Abstract. Plant cell, tissue, and organ culture is a set of techniques designed for the growth and multiplication of cells and tissues using nutrient solutions in an aseptic and controlled environment. This technology explores conditions that promote cell division and genetic reprogramming in in vitro conditions.


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