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H4
Tissue
Brain
Disease
Neuroglioma
Synonyms
H-4

Documents

H4

Cell line · Product number 300184

H4 cells are a human neuroglioma cell line derived from the central nervous system. These cells are often utilized in neurological research, particularly ...

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  • CoA
1

01General information

Description

H4 cells are a human neuroglioma cell line derived from the central nervous system. These cells are often utilized in neurological research, particularly in studies focusing on neurobiology and neuropharmacology. H4 cells provide a valuable model for understanding the molecular and cellular mechanisms of gliomas, offering insights into tumor biology, response to therapeutic agents, and the regulation of gene expression within the nervous system.

The H4 cell line is known for its robust use in experiments involving neurotoxicity and neuroprotection, serving as a tool to evaluate the effects of various substances on neuronal cells. Researchers use H4 cells to study the cellular processes involved in neurodegeneration and to screen potential neuroprotective and neuroregenerative compounds. Their consistent growth and maintenance characteristics under laboratory conditions make them a reliable resource for in vitro experiments aimed at elucidating neurological functions and disorders.

Tissue
Brain
Disease
Neuroglioma
Synonyms
  • H-4

02Classification

Organism
Human
Sex
Male
Growth properties
Adherent
Biosafety level
1

03Characteristics

Tumourigenic
No
Karyotype
Modal number = 75. range 45 = 80. Y chromosome present

04Identifiers

Citation
H4 (Cytion catalog number 300184)
NCBI TaxID
9606
Cellosaurus
CVCL_1239

05Expression and susceptibility

Protein expression
PGP9.5 postive, NeuN positive, NSE negative
Isoenzymes
G6PD, B, PGM1, 1-2, PGM3, 1, ES-D, 1, Me-2, 0, AK-1, 1, GLO-1, 2.

06Handling and culture

Culture medium
DMEM, w: 4.5 g/L Glucose, w: 4 mM L-Glutamine, w: 3.7 g/L NaHCO3, w: 1.0 mM Sodium pyruvate (Cytion article number 820300a)
Incubation atmosphere

37°C, 5% CO2, humidified atmosphere.

Supplements
Supplement the medium with 10% FBS
Dissociation reagent
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.
Seeding density
1 x 104 cells/cm2
Fluid renewal
2 to 3 times per week
Post-thaw recovery
After thawing, plate the cells at 5 x 104 cells/cm2 and allow the cells to recover from the freezing process and to adhere for at least 48 hours.
Thawing and culturing
  1. Confirm that the vial remains deeply frozen upon delivery, as cells are shipped on dry ice to maintain optimal temperatures during transit.

  2. Upon receipt, either store the cryovial immediately at temperatures below -150°C to ensure the preservation of cellular integrity, or proceed to step 3 if immediate culturing is required.

  3. For immediate culturing, swiftly thaw the vial by immersing it in a 37°C water bath with clean water and an antimicrobial agent, agitating gently for 40-60 seconds until a small ice clump remains.

  4. Perform all subsequent steps under sterile conditions in a flow hood, disinfecting the cryovial with 70% ethanol before opening.

  5. Carefully open the disinfected vial and transfer the cell suspension into a 15 ml centrifuge tube containing 8 ml of room-temperature culture medium, mixing gently.

  6. Centrifuge the mixture at 300 x g for 3 minutes to separate the cells and carefully discard the supernatant containing residual freezing medium.

  7. Gently resuspend the cell pellet in 10 ml of fresh culture medium. For adherent cells, divide the suspension between two T25 culture flasks; for suspension cultures, transfer all the medium into one T25 flask to promote effective cell interaction and growth.

  8. Adhere to established subculture protocols for continued growth and maintenance of the cell line, ensuring reliable experimental outcomes.

07Quality control and cryopreservation

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.

Freeze medium
As a cryopreservation medium, we use complete growth medium (including FBS) + 10% DMSO for adequate post-thaw viability, or CM-1 (Cytion catalog number 800100), which includes optimized osmoprotectants and metabolic stabilizers to enhance recovery and reduce cryo-induced stress.
Storage conditions

For long-term preservation, place vials in vapor-phase liquid nitrogen at about −150 to −196 °C. Storage at −80 °C is acceptable only as a short interim step before transfer to liquid nitrogen.

Shipping conditions

Cryopreserved cell lines are shipped on dry ice in validated, insulated packaging with sufficient refrigerant to maintain approximately −78 °C throughout transit. On receipt, inspect the container immediately and transfer vials without delay to appropriate storage.

08Donor

Donor age
37 years
Ethnicity
Caucasian