L-428 Cells
General information
Description | The L428 cell line is a well-established neoplastic cell line derived from the pleural effusion of a female patient diagnosed with Hodgkin's disease of the nodular sclerosing type. The establishment of this cell line has provided a valuable model for studying the cellular characteristics and molecular mechanisms underlying Hodgkin's lymphoma. L428 cells closely resemble the Reed-Sternberg (RS) and Hodgkin (H) cells, which are hallmark cells of Hodgkin's lymphoma. These cells demonstrate a unique phenotype distinct from typical B cells, T cells, and other hematopoietic cell types, contributing to ongoing debates about the exact cellular origin of RS and H cells. The L428 cell line exhibits several distinctive characteristics, including aneuploidy and the presence of multiple structural and numerical chromosomal abnormalities, which are typical markers of its neoplastic nature. These cells lack surface or cytoplasmic immunoglobulins (Igs), despite their derivation from a lymphoid malignancy, which suggests significant differentiation from normal lymphoid cells. The absence of Epstein-Barr Virus (EBV) antigens, such as EBNA and VCA, further distinguishes L428 from other EBV-positive Hodgkin's lymphoma cell lines. The cells also lack lysozyme, peroxidase, and chloracetate esterase activity, reinforcing their distinction from myeloid cells, monocytes, or macrophages. In terms of morphology, L428 cells exhibit a range of sizes, from small mononuclear cells to large multinucleated cells, with some cells displaying villous projections on their membranes. The cells are also notable for their large, often kidney-shaped nucleoli. Functionally, L428 cells express Ia-like antigens and T-cell receptors but are devoid of other common lymphoid and myeloid markers. This unique immunophenotype, combined with the chromosomal and morphological features, supports the classification of L428 as a model of Hodgkin's lymphoma, particularly for studying the biology of RS and H cells. The L428 cell line has been used extensively in research to explore the pathogenesis of Hodgkin's disease and to investigate potential therapeutic targets. Its ability to proliferate in vitro and its unique properties make it a critical resource for advancing the understanding of this complex hematological malignancy. |
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Organism | Human |
Tissue | Pleural effusion |
Disease | Hodgkin lymphoma |
Synonyms | L-428, L 428 |
Characteristics
Age | 37 years |
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Gender | Female |
Ethnicity | Caucasian |
Morphology | Round cells |
Cell type | Lymphoblast |
Growth properties | Suspension |
Identifiers / Biosafety / Citation
Citation | L428 (Cytion catalog number 300200) |
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Biosafety level | 1 |
Expression / Mutation
Handling
Culture Medium | RPMI 1640, w: 2.1 mM stable Glutamine, w: 2.0 g/L NaHCO3 (Cytion article number 820700a) |
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Medium supplements | Supplement the medium with 10% FBS, 1 mM sodium pyruvate, 1% NEAA |
Subculturing | Maintain cultures by periodically adding or replacing the medium. Initiate cultures with a density of 2 x 10^5 cells/ml and keep the cell concentration within the range of 1 x 10^5 to 1 x 10^6 cells/ml for optimal growth. |
Seeding density | 1 x 10^5 cells/ml |
Fluid renewal | Every 3 days |
Freezing recovery | Fast |
Freeze medium | CM-1 (Cytion catalog number 800100) |
Handling of cryopreserved cultures |
|
Quality control / Genetic profile / HLA
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 |
CSF1PO: 10,13
D13S317: 14,14
D16S539: 11,12
D5S818: 11,12
D7S820: 11,11
TH01: 7,9.3
TPOX: 8,9
vWA: 15
D3S1358: 14,18
D21S11: 31.2,31.2
D18S51: 14,14
Penta E: 10,17
Penta D: 8,9
D8S1179: 14,14
FGA: 19,25
|
HLA alleles |
A*: 03:01:01
B*: 35:03:01
C*: 04:01:01
DRB1*: 12:01:01
DQA1*: 05:05:01
DQB1*: 03:01:01
DPB1*: 04:01:01
E: 01:03:02
|
Required products
Initially designed to support the growth of human leukemic cells in both suspension and monolayer cultures, RPMI 1640 Medium has evolved through modifications by researchers and commercial suppliers to become suitable for a diverse range of mammalian cells. It is exceptionally compatible with cell lines such as HeLa, Jurkat, MCF-7, PC12, PBMC, astrocytes, and carcinomas.
RPMI 1640 Medium stands apart from other cell culture media due to its unique composition. It contains a substantial amount of phosphate, amino acids, and vitamins. Notably, it encompasses biotin, vitamin B12, and PABA, absent in Eagle's Minimal Essential Medium or Dulbecco's Modified Eagle Medium. Moreover, RPMI 1640 Medium exhibits significantly elevated concentrations of vitamins inositol and choline. However, it does not contain proteins, lipids, or growth factors. Consequently, supplementation with 10% Fetal Bovine Serum (FBS) is commonly required to provide optimal conditions for cell growth.
The buffering system of RPMI 1640 Medium relies on sodium bicarbonate (2.0 g/L) and necessitates a 5-10% CO2 environment to maintain a physiologically appropriate pH. The inclusion of the reducing agent glutathione further distinguishes this medium from others.
The unique composition of this RPMI formulation comprises 2.1 mM of stable Glutamine, 2.0 grams per liter of NaHCO3, and phenol red.
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 nitrate x 4H2O
100,00
Magnesium sulfate anhydrous
48,83
Potassium chloride
400,00
Sodium chloride
5,950.00
di-Sodium hydrogen phosphate
800,49
Other Components
D(+)-Glucose anhydrous
2,000.00
Glutathione (red.)
1,00
Phenol red
5,00
NaHCO3
2,000.00
Amino Acids
L-Arginine x HCl
241,86
L-Asparagine x H2O
56,82
L-Aspartic acid
20,00
L-Cystine x 2HCl
65,19
L-Alanyl-L-Glutamine
447,00
L-Glutamic acid
20,00
Glycine
10,00
L-Histidine x HCl x H2O
20,27
L-Hydroxyproline
20,00
L-Isoleucine
50,00
L-Leucine
50,00
L-Lysine x HCl
40,00
L-Methionine
15,00
L-Phenylalanine
15,00
L-Proline
20,00
L-Serine
30,00
L-Threonine
20,00
L-Tryptophan
5,00
L-Tyrosine x 2Na
28,83
L-Valine
20,00
Vitamins
p-Aminobenzoic acid
1,00
D-(+)-Biotin
0,20
D-Calcium pantothenate
0,25
Choline chloride
3,00
Folic acid
1,00
myo-Inositol
35,00
Nicotinamide
1,00
Pyridoxine x HCl
1,00
Riboflavin
0,20
Thiamine x HCl
1,00
Vitamin B12
0.005
Initially designed to support the growth of human leukemic cells in both suspension and monolayer cultures, RPMI 1640 Medium has evolved through modifications by researchers and commercial suppliers to become suitable for a diverse range of mammalian cells. It is exceptionally compatible with cell lines such as HeLa, Jurkat, MCF-7, PC12, PBMC, astrocytes, and carcinomas.
RPMI 1640 Medium stands apart from other cell culture media due to its unique composition. It contains a substantial amount of phosphate, amino acids, and vitamins. Notably, it encompasses biotin, vitamin B12, and PABA, absent in Eagle's Minimal Essential Medium or Dulbecco's Modified Eagle Medium. Moreover, RPMI 1640 Medium exhibits significantly elevated concentrations of vitamins inositol and choline. However, it does not contain proteins, lipids, or growth factors. Consequently, supplementation with 10% Fetal Bovine Serum (FBS) is commonly required to provide optimal conditions for cell growth.
The buffering system of RPMI 1640 Medium relies on sodium bicarbonate (2.0 g/L) and necessitates a 5-10% CO2 environment to maintain a physiologically appropriate pH. The inclusion of the reducing agent glutathione further distinguishes this medium from others.
This RPMI 1640 medium contains 4.5 grams per liter of glucose.
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 nitrate x 4H2O
100,00
Magnesium sulfate anhydrous
48,83
Potassium chloride
400,00
Sodium chloride
5450,00
di-Sodium hydrogen phosphate
800,49
Other Components
D(+)-Glucose anhydrous
4500,00
Glutathione (red.)
1,00
HEPES
2383,00
Phenol red
5,00
Sodium pyruvate
110,00
Amino Acids
L-Arginine x HCl
241,86
L-Asparagine x H2O
56,82
L-Aspartic acid
20,00
L-Cystine x 2HCl
65,19
L-Glutamine
300,00
L-Glutamic acid
20,00
Glycine
10,00
L-Histidine x HCl x H2O
20,27
L-Hydroxyproline
20,00
L-Isoleucine
50,00
L-Leucine
50,00
L-Lysine x HCl
40,00
L-Methionine
15,00
L-Phenylalanine
15,00
L-Proline
20,00
L-Serine
30,00
L-Threonine
20,00
L-Tryptophan
5,00
L-Tyrosine x 2Na
28,83
L-Valine
20,00
Vitamins
p-Aminobenzoic acid
1,00
D-(+)-Biotin
0,20
D-Calcium pantothenate
0,25
Choline chloride
3,00
Folic acid
1,00
myo-Inositol
35,00
Nicotinamide
1,00
Pyridoxine x HCl
1,00
Riboflavin
0,20
Thiamine x HCl
1,00
Vitamin B12
0,01
NaHCO3
1500,00
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