MRC-5 Cells
Insights on MRC-5 cells
Description | MRC-5 cells, a human lung fibroblast cell line derived from the lung tissue of a 14-week-old male fetus in 1966, are extensively utilized in the production of certain vaccines, including those for hepatitis A, polio, rabies, and more. The susceptibility to various human viruses, notably human poliovirus 1, herpes simplex virus, and vesicular stomatitis virus underscores the role of MRC5 cells in the discovery of antivirals, viral vaccines, vaccine safety and virus replication. MRC-5 and WI-38 cell lines are still used in producing vaccines for varicella, rubella, hepatitis A, and a version of rabies vaccine today. Recently, MRC-5 cells were modified to express the ACE2 receptor, and have been key in SARS research. The modified MRC5 human ace2 cells allow scientists to study how the SARS-CoV virus enters and replicates in host cells. This work has been vital for understanding the virus's behavior and developing targeted antiviral agents and treatments. The MRC5 fetal cell line's utility extends beyond vaccine production to include potential roles in cancer research, with the cell line being employed in studies exploring the tumor microenvironment and cancer cell interactions, owing to their capability to differentiate into multiple cell types, including osteocytes and chondrocytes. This has led to speculation about their similarity to mesenchymal stem cells (MSCs), given their fibroblast-like morphology and maintenance of a normal diploid karyotype over extensive in vitro expansion. |
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Organism | Human |
Tissue | Lung |
Applications | Vaccine production |
Synonyms | MRC5, MRC 5, MRCV, MRC-V, Medical Research Council cell strain-5 |
Details
Age | Fetus |
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Gender | Male |
Cell type | Fibroblast |
Growth properties | Adherent |
Documentation
Citation | MRC-5 (Cytion catalog number 300395) |
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Biosafety level | 1 |
Genetic characteristics of the MRC5 cell line
Virus susceptibility | Not susceptible to SARS coronavirus 2 (SARS-CoV-2) infection (COVID-19) |
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Karyotype | MRC5 is a diploid cell line with a modal chromosome number of 46. |
Handling
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) |
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Medium supplements | Supplement the medium with 10% FBS |
Passaging solution | Accutase |
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. |
Freeze medium | CM-1 (Cytion catalog number 800100) |
Handling of cryopreserved cultures |
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Quality control of MRC-5 fetal 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. |
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STR profile |
Amelogenin: x,y
CSF1PO: 11,12
D13S317: 11,14
D16S539: 9,11
D5S818: 11,12
D7S820: 10,11
TH01: 8
TPOX: 8
vWA: 15
D3S1358: 15,17
D21S11: 31.2
D18S51: 15,21
Penta E: 12,16
Penta D: 12
D8S1179: 13
FGA: 21,23
D6S1043: 11,19
D2S1338: 20
D12S391: 20,22
D19S433: 14,15
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HLA alleles |
A*: 02:01:01, 29:02:01
B*: 07:02:01, 44:02:01
C*: 05:01:01, 07:02:01
DRB1*: 04:08:01, 15:01:01G
DQA1*: 01:02:01, 03:03:01
DQB1*: 03:01:01, 06:02:01
DPB1*: 04:01:01
E: 01:01:01
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Required products
- 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.