HCM-BROD-0012-C71 (ATCC® PDM-19)

Organism: Homo sapiens, human  /  Tissue: brain  /  Disease: recurrent glioblastoma

Permits and Restrictions

View Permits

Organism Homo sapiens, human
Tissue brain
Product Format frozen 1.0 mL
Morphology neuronal
Culture Properties adherent
Biosafety Level 1

Biosafety classification is based on U.S. Public Health Service Guidelines, it is the responsibility of the customer to ensure that their facilities comply with biosafety regulations for their own country.

Disease recurrent glioblastoma
Age See associated clinical data for patient profile information, if available.
https://portal.gdc.cancer.gov/  
https://hcmi-searchable-catalog.nci.nih.gov/model/HCM-BROD-0012-C71

Gender female
Ethnicity See associated clinical data for patient profile information, if available.
https://portal.gdc.cancer.gov/  
https://hcmi-searchable-catalog.nci.nih.gov/model/HCM-BROD-0012-C71

Applications Basic research, compound screening
Storage Conditions liquid nitrogen vapor phase
Images
Clinical Data ICD-10-CM code: C71, malignant neoplasm of cerebrum; Reccurent glioblastoma
https://hcmi-searchable-catalog.nci.nih.gov/model/HCM-BROD-0012-C71

Comments ICD-10-CM code: C71, malignant neoplasm of cerebrum.
Next-generation cancer model from the Human Cancer Models Initiate (HCMI). Refer to the following websites for additional information on this model including protocols, clinical information, and bioinformatics data.

https://ocg.cancer.gov/programs/hcmi/resources
https://portal.gdc.cancer.gov/  
https://hcmi-searchable-catalog.nci.nih.gov/model/HCM-BROD-0012-C71

Complete Growth Medium

NeuroCult NS-A Basal Medium (StemCell Technologies #05750) with NS-A Proliferation Supplement (StemCell Technologies #05754) + 20 ng/mL EGF (StemCell Technologies #78003.1) + 20 ng/mL bFGF (Peprotech #AF-100-15) + 2 µg/mL Heparin (StemCell Technologies #07980).

Prepare media according to the manufacturer’s instructions: Stem Cell Technologies Catalog #5751

Subculturing

Culture vessels must be pre-coated with laminin prior to seeding with cells.

Coating procedure (perform all steps in a biosafety cabinet):

  1. Thaw (Corning #354232) laminin on ice.
  2. Transfer 10 mL of D-PBS (ATCC 30-2200) to a 15 mL conical tube.
  3. Add 100 µL stock laminin to the 15 mL conical tube with D-PBS and mix thoroughly.
  4. Added diluted laminin to vessels at a volume of approximately 1 mL per 10 cm2. For example, use 7.5 mL diluted laminin for a 75 cm2 flask.
  5. Tilt plate to ensure entire surface is coated.
  6. Place in a humid cell culture incubator at 37oC for 1-24 hours.

Immediately prior to seeding cells, aspirate off the laminin coating solution and discard. Do not allow the plates to dry out.

  1. Passage cells when the culture has reached approximately 70% to 80% confluence.
  2. Pre-coat new culture vessels with laminin, if necessary.
  3. Warm Accutase and complete growth media to room temperature.
  4. For each flask, carefully aspirate the spent media without disturbing the monolayer.
  5. Rinse the cell layer one time with 3 to 5 mL D-PBS (ATCC 30-2200) to remove residual medium.
  6. Add room temperature Accutase (1 to 2 mL for every 25 cm2) to each flask.
  7. Gently rock each flask to ensure complete coverage of the Accutase solution over the cells, and then aspirate the excess fluid off of the monolayer.
  8. Observe the cells under the microscope.
  9. When the majority of cells appear to have detached (typically 2-5 minutes), quickly add an equal volume of the complete growth medium to each flask.
  10. Transfer the dissociated cells to a sterile centrifuge tube.
  11. Centrifuge the cells at 200 x g for 5 minutes.
  12. Aspirate neutralized dissociation solution from the cell pellet and resuspend the cells in 2 to 8 mL fresh, pre-warmed, complete growth medium.
  13. Count the cells and seed new culture flasks at a density of 5 x 104 viable cells per cm2. Prior to seeding, aspirate the coating and discard the coating laminin solution from the vessel.
  14. Place newly seeded flasks in a 37°C, 5% CO2 incubator for at least 24 to 48 hours before processing the cells further.
  15. Perform a complete medium change every 3-4 days.
Cryopreservation Complete growth media containing 10% DMSO (ATCC 4-X).
Culture Conditions
Atmosphere: air, 95%; carbon dioxide (CO2), 5%
Temperature: 37°C
Cells per Vial ≥ 1 x 106 cells
Volume 1.0 mL
STR Profile
Amelogenin: X
CSF1PO: 7,11
D13S317: 8,12
D16S539: 12
D5S818: 11,13
D7S820: 10,11
THO1: 9,9.3
TPOX: 8,11
vWA: 15
Sterility Tests Bacteria, yeast and fungi: No growth
Mycoplasma: No growth
Viral Testing Hepatitis B: None detected
Cytomegalovirus: None detected
Human immunodeficiency virus: None detected
Epstein-Barr virus: None detected
Human papillomavirus: None detected
Name of Depositor Broad Institute
Year of Origin 2018
Notice: Necessary PermitsPermits

These permits may be required for shipping this product:

  • Customers located in the state of Hawaii will need to contact the Hawaii Department of Agriculture to determine if an Import Permit is required. A copy of the permit or documentation that a permit is not required must be sent to ATCC in advance of shipment.
Basic Documentation
Other Documentation
FAQ's
  1. HCMI - Human Cancer Models Initiative


    Date Updated: 2/4/2019

  2. 3D Models for commercial use
    Please refer to the “Restrictions” section on the product page of the HCMI model page of interest. It will list any product use restrictions or licenses required.
    Date Updated: 6/5/2019
  3. 3D Models sequencing and clinical data
    Visit the HCMI Searchable Catalog page for the model of interest to see available model specific data. Link to the catalog:
    Date Updated: 6/5/2019
  4. Neurosphere models
    Neurospheres are aggregates of brain-derived cell types that are propagated in culture as multicellular spheres that grow in suspension.
    Date Updated: 6/5/2019
  5. Conditionally reprogrammed cells (CRC) growth medium
    CRC media is commercially available from Propagenix. Their website is:
    Date Updated: 6/5/2019
  6. Conditionally reprogrammed cells - CRC
    CRC technology was developed at Georgetown University and permits the expansion of patient derived epithelial cells typically utilizing a combination of co-culture with feeder cells and media con...
    Date Updated: 6/5/2019