HeLa Cells
History of Henrietta Lacks' cells
Description | HeLa cells, derived from the cervical cancer cells of Henrietta Lacks, are an immortal cell line widely employed in biomedical research. The human cell line Hela has significantly contributed to significant research advances and continues to play a pivotal role in laboratories worldwide. In 1951, Henrietta Lacks, a young mother of five, sought medical attention at The Johns Hopkins Hospital for vaginal bleeding, where Dr. Howard Jones identified a significant malignant tumor on her cervix. At that time, the Johns Hopkins Medicine Institute was among the few institutions offering medical care to impoverished African Americans. Henrietta Lacks underwent radium treatment for her cervical cancer, the leading therapy available then. During her treatment, a biopsy was conducted, and a sample of her cancerous cells was sent to Dr. George Otto Gey's lab. Dr. Gey had been attempting to cultivate cells from cervical cancer patients of diverse backgrounds, but without success until Henrietta's cells, which were the first cells to proliferate continuously, a discovery that set them apart from all previous samples. Henrietta Lacks' cervical carcinoma was later found to have been caused by the Human papillomavirus (HPV). HPV is a common virus that can lead to cervical cancer among other diseases. Research on HeLa cells has significantly contributed to understanding the role of HPV in cervical cancer, leading to the development of preventive HPV vaccines, which have had a profound impact on reducing the incidence of HPV-related cancers. These extraordinary cells, termed "HeLa" cells after Henrietta Lacks' initials, have since become instrumental in medical research. They have enabled scientists to investigate cancer cell growth, the impact of various substances, and the workings of viruses, significantly contributing to medical advancements, including the development of vaccines for polio and COVID-19, without the ethical concerns of direct human experimentation. HeLa cells are widely used for gene function studies, recombinant protein production, and gene therapy due to their high transfection efficiency and susceptibility to viral infections. They are pivotal in researching viral behaviors, including replication and pathogenesis, and have played a key role in Hepatitis B research by expressing viral proteins and aiding in the development of diagnostic tests and vaccines, thereby significantly advancing global health measures. HeLa cells continue to be an invaluable resource for ongoing research in medicine and science. The significance of HeLa cells and other immortal cell lines cannot be overstated, as they continue to shape the field of medicine and infectious disease research, and they represent a lasting legacy of Henrietta Lacks and her contributions to scientific advancement. |
---|---|
Organism | Human |
Tissue | Cervix |
Disease | Adenocarcinoma |
Applications | Transfection host |
Synonyms | HELA, Hela, He La, He-La, Henrietta Lacks cells, Helacyton gartleri |
Details
Age | 30 years |
---|---|
Gender | Female |
Ethnicity | African American |
Morphology | Epithelial-like |
Growth properties | Adherent |
Documentation
Citation | HeLa (Cytion catalog number 300194) |
---|---|
Biosafety level | 1 |
Genetic profile
Isoenzymes | G6PD, A |
---|---|
Virus susceptibility | Human adenovirus 3, Encephalomyocarditis virus, Human poliovirus 1, Human poliovirus 2, Human poliovirus 3 |
Reverse transcriptase | Negative |
Products | Keratin, Lysophosphatidylcholine (lyso-PC) induces AP-1 activity and c-jun N-terminal kinase activity (JNK1) by a protein kinase C-independent pathway |
Karyotype | The HeLa cell line, with its complex karyotype featuring a high degree of aneuploidy and structural rearrangements, is known for its rapid growth and longevity in culture. HeLa cells typically exhibit 82 chromosomes, although the range can vary from 70 to 164. Notably, 98% of HeLa cells possess a small telocentric chromosome, and 100% exhibit aneuploidy in a substantial number of cells examined. These chromosomal abnormalities underpin their fast growth and immortality, along with their association with cervical cancer and other cancerous cells. |
Handling the Hela cell line
Culture Medium | EMEM, w: 2 mM L-Glutamine, w: 1.5 g/L NaHCO3, w: EBSS, w: 1 mM Sodium pyruvate, w: NEAA (Cytion article number 820100c) |
---|---|
Medium supplements | Supplement the medium with 10% FBS |
Passaging solution | Accutase |
Doubling time | 28 to 36 hours |
Subculturing | Remove the old medium from the adherent cells and wash them with PBS that lacks calcium and magnesium. For T25 flasks, use 3-5 ml of PBS, and for T75 flasks, use 5-10 ml. Then, cover the cells completely with Accutase, using 1-2 ml for T25 flasks and 2.5 ml for T75 flasks. Let the cells incubate at room temperature for 8-10 minutes to detach them. After incubation, gently mix the cells with 10 ml of medium to resuspend them, then centrifuge at 300xg for 3 minutes. Discard the supernatant, resuspend the cells in fresh medium, and transfer them into new flasks that already contain fresh medium. |
Split ratio | A ratio of 1:2 to 1:6 is recommended |
Seeding density | 1 x 10^4 cells/cm^2 |
Fluid renewal | 2 to 3 times per week |
Freezing recovery | After thawing, plate the cells at 2 to 3 x 10^4 cells/cm^2 and allow the cells to recover from the freezing process and to adhere for at least 24 to 48 hours. |
Freeze medium | CM-1 (Cytion catalog number 800100) or CM-ACF (Cytion catalog number 806100) |
Handling of cryopreserved cultures |
|
Quality assurance of Hela cells
Sterility | Mycoplasma contamination is excluded using both PCR-based assays and luminescence-based mycoplasma detection methods. To ensure there is no bacterial, fungal, or yeast contamination, cell cultures are subjected to daily visual inspections. |
---|---|
STR profile |
Amelogenin: x,x
CSF1PO: 9,1
D13S317: 12,13.3
D16S539: 9,1
D5S818: 11,12
D7S820: 8,12
TH01: 7
TPOX: 8,12
vWA: 16,18
D3S1358: 15,18
D21S11: 27,28
D18S51: 16
Penta E: 7,17
Penta D: 8,15
D8S1179: 12,13
FGA: 18,21
D6S1043: 18
D2S1338: 17
D12S391: 20,25
D19S433: 13,14
|
HLA alleles |
A*: 68:02:01
B*: 15:03:01
C*: 12:03:01
DRB1*: 01:02:01
DQA1*: 01:01:02
DQB1*: 05:01:01
DPB1*: 01:01:01
E: 01:03:02
|
Required products
Key features of Freeze Medium CM-1 include:
Broad Compatibility: Effective for a wide range of cell types, including primary cells, stem cells, and established cell lines.
High Viability: Optimized to maximize post-thaw cell recovery and viability, ensuring reliable experimental outcomes.
Ready-to-Use: Conveniently prepared and sterilized for immediate application, reducing preparation time and risk of contamination.
Enhanced Stability: Maintains consistent performance under standard cryopreservation conditions, ensuring reproducible results.
Long Shelf Life: CM-1 is a serum-containing, ready-to-use cryopreservation medium that can be stored in the refrigerator for up to one year.
Using CM-1 for Freezing Cells
To use CM-1 for freezing both adherent and suspension cells, follow these steps:
For adherent cells, wash and dissociate them from the culture substrate. For suspension cells, proceed directly to the next step.
Count the cells to ensure they are at the proper concentration.
Centrifuge the cells to pellet them, then resuspend in CM-1 freeze medium.
Transfer the resuspended cells into cryovials.
Use a slow-freezing method before transferring the cells to long-term storage.
🥶 Method
🔍 Description
💡 Steps
❄️
Manual Freezing
A step-by-step method involving gradual temperature reduction to ensure cell viability.
1️⃣ Place cells in freeze medium in a 4°C freezer for 40 minutes.
2️⃣ Transfer to a -80°C freezer for 24 hours.
3️⃣ Store cells in liquid nitrogen for long-term preservation.
🧊
Using Mr. Frosty
A convenient device that allows for controlled freezing rates without electrical power.
1️⃣ Prepare cells in cryovials with freeze medium.
2️⃣ Place cryovials in Mr. Frosty container.
3️⃣ Store at -80°C for 24 hours before transferring to liquid nitrogen.
🧬
Controlled-Rate Freezer
A high-precision freezer by Thermo Fisher or other manufacturers designed for controlled temperature reduction.
1️⃣ Program the device to gradually decrease the temperature.
2️⃣ Place prepared cells in the freezer.
3️⃣ After the freezing cycle, transfer cells to liquid nitrogen.
Store the cryovials at temperatures below -130°C or in liquid nitrogen for long-term preservation.
Ingredients
Contains FBS, DMSO, Glucose, Salts
Buffering capacity: pH = 7.2 to 7.6
Cytion’s Freeze Medium CM-1 offers a reliable solution for cryopreservation, ensuring high cell viability and functionality post-thaw for a wide range of research applications.
This EMEM medium consists of 2 mM L-Glutamine, 1.5 g/L NaHCO3, EBSS, 1 mM Sodium pyruvate, and NEAA.
What's in EMEM?
EMEM is a modified version of Eagle's minimum essential medium, containing Earle's Balanced Salt Solution, non-essential amino acids, L-glutamine, sodium pyruvate, and sodium bicarbonate. It's important to note that this reduced level of sodium bicarbonate (NaHCO3, 1.5 g/L) is intended for use in 5% CO2 in the air. To maintain its effectiveness, storing the medium at two °C to 8°C in the dark when not in use is recommended.
What is EMEM used for?
Eagle's minimal essential medium (EMEM) is a cell culture medium that can maintain cells in tissue culture. The medium contains higher concentrations of amino acids, allowing for a more accurate approximation of the protein composition of cultured mammalian cells. EMEM may be used to cultivate various cells, including fibroblasts, human liver cancer cell line (HepG2) cells and human fetal brain progenitor-derived astrocyte cells (PDA). It is typically used in the presence of fetal bovine serum (FBS), calf, or horse sera.
How is EMEM different from other cell culture media?
While EMEM and Dulbecco's modified Eagle's medium (DMEM) share some similarities, they also differ. Both media lack protein and contain the amino acids, salts, glucose, and vitamins required to provide a cell with energy and maintain it in tissue culture. However, the DMEM formulation is modified to contain up to four times more vitamins and amino acids and two to four times more glucose than EMEM. It's worth noting that EMEM is also different from the original MEM formulation.
Quality control
pH = 7.2 +/
- 0.02 at 20-25°C.
Each lot has been tested for sterility and absence of mycoplasma and bacteria.
Maintenance
Keep refrigerated at +2°C to +8°C in the dark. Freezing and warming up to +37° C minimize the quality of the product.
Do not heat the medium to more than 37° C or use uncontrollable sources of heat (e.g., microwave appliances).
If only a part of the medium is to be used, remove this amount from the bottle and warm it up at room temperature.
Shelf life for any medium except for the basic medium is 8 weeks from the date of manufacture.
Composition
Components
mg/L
Inorganic Salts
Calcium chloride x 2H2O
264,92
Magnesium sulfate
97,67
Potassium chloride
400,00
Sodium chloride
6,800.00
Sodium dihydrogen phosphate x H2O
140,00
Other Components
D(+)-Glucose
1,000.00
Phenol red
10,00
Sodium pyruvate
110,00
NaHCO3
1,500.00
Amino Acids
L-Alanine
8,90
L-Arginine x HCl
126,00
L-Asparagine x H2O
13,20
L-Aspartic acid
13,30
L-Cystine
24,00
L-Glutamine
292,30
L-Glutamic acid
14,70
Glycine
7,50
L-Histidine x HCl x H2O
42,00
L-Isoleucine
52,00
L-Leucine
52,00
L-Lysine x HCl
72,50
L-Methionine
15,00
L-Phenylalanine
32,00
L-Proline
11,50
L-Serine
10,50
L-Threonine
48,00
L-Tryptophan
10,00
L-Tyrosine
36,00
L-Valine
46,00
Vitamins
D-Calcium pantothenate
1,00
Choline chloride
1,00
Folic acid
1,00
myo-Inositol
2,00
Nicotinamide
1,00
Pyridoxal x HCl
1,00
Riboflavin
0,10
Thiamine x HCl
1,00
- A Gentle Alternative to Trypsin
Accutase is a cell detachment solution that is revolutionizing the cell culture industry. It is a mix of proteolytic and collagenolytic enzymes that mimics the action of trypsin and collagenase. Unlike trypsin, Accutase does not contain any mammalian or bacterial components and is much gentler on cells, making it an ideal solution for the routine detachment of cells from standard tissue culture plasticware and adhesion coated plasticware. In this blog post, we will explore the benefits and uses of Accutase and how it is changing the game in cell culture.
Advantages of Accutase
Accutase has several advantages over traditional trypsin solutions. Firstly, it can be used whenever gentle and efficient detachment of any adherent cell line is needed, making it a direct replacement for trypsin. Secondly, Accutase works extremely well on embryonic and neuronal stem cells, and it has been shown to maintain the viability of these cells after passaging. Thirdly, Accutase preserves most epitopes for subsequent flow cytometry analysis, making it ideal for cell surface marker analysis.
Additionally, Accutase does not need to be neutralized when passaging adherent cells. The addition of more media after the cells are split dilutes Accutase so it is no longer able to detach cells. This eliminates the need for an inactivation step and saves time for cell culture technicians. Finally, Accutase does not need to be aliquoted, and a bottle is stable in the refrigerator for 2 months.
Applications of Accutase
Accutase is a direct replacement for trypsin solution and can be used for the passaging of cell lines. Additionally, Accutase performs well when detaching cells for the analysis of many cell surface markers using flow cytometry and for cell sorting. Other downstream applications of Accutase treatment include analysis of cell surface markers, virus growth assay, cell proliferation, tumor cell migration assays, routine cell passage, production scale-up (bioreactor), and flow cytometry.
Composition of Accutase
Accutase contains no mammalian or bacterial components and is a natural enzyme mixture with proteolytic and collagenolytic enzyme activity. It is formulated at a much lower concentration than trypsin and collagenase, making it less toxic and gentler, but just as effective.
Efficiency of Accutase
Accutase has been shown to be efficient in detaching primary and stem cells and maintaining high cell viability compared to animal origin enzymes such as trypsin. 100% of cells are recovered after 10 minutes, and there is no harm in leaving cells in Accutase for up to 45 minutes, thanks to autodigestion of Accutase.
In summary
In conclusion, Accutase is a powerful solution that is changing the game in cell culture. With its gentle nature, efficiency, and versatility, Accutase is the ideal alternative to trypsin. If you are looking for a reliable and efficient solution for cell detachment, Accutase is the solution for you.
Phosphate-buffered saline (PBS) is a versatile buffer solution used in many biological and chemical applications, as well as tissue processing. Our PBS solution is formulated with high-quality ingredients to ensure a constant pH during experiments. The osmolarity and ion concentrations of our PBS solution are matched to those of the human body, making it isotonic and non-toxic to most cells.
Composition of our PBS Solution
Our PBS solution is a pH-adjusted blend of ultrapure-grade phosphate buffers and saline solutions. At a 1X working concentration, it contains 137 mM NaCl, 2.7 mM KCl, 8 mM Na2HPO4, and 2 mM KH2PO4. We have chosen this composition based on CSHL protocols and Molecular cloning by Sambrook, which are well-established standards in the research community.
Applications of our PBS Solution
Our PBS solution is ideal for a wide range of applications in biological research. Its isotonic and non-toxic properties make it perfect for substance dilution and cell container rinsing. Our PBS solution with EDTA can also be used to disengage attached and clumped cells. However, it is important to note that divalent metals such as zinc cannot be added to PBS as this may result in precipitation. In such cases, Good's buffers are recommended. Moreover, our PBS solution has been shown to be an acceptable alternative to viral transport medium for the transport and storage of RNA viruses, such as SARS-CoV-2.
Storage of our PBS Solution
Our PBS solution can be stored at room temperature, making it easy to use and access.
To sum up
In summary, our PBS solution is an essential component in many biological and chemical experiments. Its isotonic and non-toxic properties make it suitable for numerous applications, from cell culture to viral transport medium. By choosing our high-quality PBS solution, researchers can optimize their experiments and ensure accurate and reliable results.
Composition
Components
mg/L
Inorganic Salts
Potassium chloride
200,00
Potassium dihydrogen phosphate
200,00
Sodium chloride
8,000.00
di-Sodium hydrogen phosphate anhydrous
1,150.00