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HCM-BROD-0830-C71

PDM-503

A patient-derived next-generation cancer model generated by the Human Cancer Models Initiative (HCMI). HCM-BROD-0830-C71 (ATCC No. PDM-503) was isolated from a recurrent glioblastoma tumor from brain tissue. This tumor-derived model can be used in basic research and pharmacological screening applications. Data for the parental tumor and the tumor-derived organoid models are available at the GDC. Additional molecular characterizations may be available at the GDC. Additional controlled data may be available via dbGaP.
Product category
Human cells
Product type
Cell model
Organism
Homo sapiens, human
Morphology
neuronal
Tissue
Brain
Disease
Glioblastoma; Recurrent
Applications
Cancer research
Neuroscience
Product format
Frozen
Storage conditions
Vapor phase of liquid nitrogen
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Documentation

ATCC determines the biosafety level of a material based on our risk assessment as guided by the current edition of Biosafety in Microbiological and Biomedical Laboratories (BMBL), U.S. Department of Health and Human Services. It is your responsibility to understand the hazards associated with the material per your organization’s policies and procedures as well as any other applicable regulations as enforced by your local or national agencies.

ATCC highly recommends that appropriate personal protective equipment is always used when handling vials. For cultures that require storage in liquid nitrogen, it is important to note that some vials may leak when submersed in liquid nitrogen and will slowly fill with liquid nitrogen. Upon thawing, the conversion of the liquid nitrogen back to its gas phase may result in the vial exploding or blowing off its cap with dangerous force creating flying debris. Unless necessary, ATCC recommends that these cultures be stored in the vapor phase of liquid nitrogen rather than submersed in liquid nitrogen.

Required Products

These products are vital for the proper use of this item and have been confirmed as effective in supporting functionality. If you use alternative products, the quality and effectiveness of the item may be affected.

Detailed product information

General

Specific applications

Basic research, compound screening

Characteristics

Cells per vial
≥ 1.0 x 106
Volume
1.0 mL
Growth properties
2D adherent
Clinical data

ICD-10-CM code: C71, malignant neoplasm of cerebrum; Recurrent glioblastoma

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-0830-C71
Comments
Next-generation cancer model from the Human Cancer Models Initiative (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-0830-C71
 

If use of this culture results in a scientific publication, it should be cited in that manuscript in the following manner: HCM-BROD-0830-C71 (ATCC® PDM-503).

Additionally, please acknowledge the HCMI as follows:  “We used models and data derived by the Human Cancer Models Initiative (HCMI) https://ocg.cancer.gov/programs/HCMI; dbGaP accession number phs001486.”
 

Handling information

Complete 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

Temperature
37°C
Atmosphere
95% Air, 5% CO2
Handling procedure

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 uL 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 75cm2 flask.
  5. Tilt plate to ensure entire surface is coated.
  6. Place in a humid cell culture incubator at 37C 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.

To ensure the highest level of viability, thaw the vial and initiate the culture as soon as possible upon receipt. If upon arrival, continued storage of the frozen culture is necessary, it should be stored in liquid nitrogen vapor phase and not at -70°C. Storage at -70°C will result in loss of viability. 

  1. Thaw the vial by gentle agitation in a 37°C water bath. To reduce the possibility of contamination, keep the O-ring and cap out of the water. Thawing should be rapid (approximately 2 minutes).
  2. Remove the vial from the water bath as soon as the contents are thawed, and decontaminate by dipping in or spraying with 70% ethanol. All of the operations from this point on should be carried out under strict aseptic conditions.
  3. Transfer the vial contents to a centrifuge tube containing 9.0 mL complete culture medium. and spin at approximately 200 x g for 5 minutes.
  4. Resuspend cell pellet with the recommended complete medium (see the specific batch information for the culture recommended dilution ratio). It is important to avoid excessive alkalinity of the medium during recovery of the cells. It is suggested that, prior to the addition of the vial contents, the culture vessel containing the complete growth medium be placed into the incubator for at least 15 minutes to allow the medium to reach its normal pH (7.0 to 7.6). pH (7.0 to 7.6).
  5. Incubate the culture at 37°C in a suitable incubator. A 5% CO2 in air atmosphere is recommended if using the medium described on this product sheet.

     

Subculturing procedure

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 cat#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 37°C 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 (StemCell Technologies cat# 07920) 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.


Reagents for cryopreservation

Complete growth media containing 10% DMSO (ATCC 4-X)

Quality control specifications

Bacterial and fungal testing
Not detected
Mycoplasma contamination
Not detected
Virus testing
Cytomegalovirus (CMV): Not detected
Epstein-Barr virus (EBV): Not detected
Hepatitis B virus (HBV): Not detected
Human Immunodeficiency virus (HIV): Not detected
Human papillomavirus (HPV): Not detected

History

Depositors
Broad Institute
Year of origin
2021
Special collection
Human Cancer Models Initiative (HCMI)

Legal disclaimers

Intended use
This product is intended for laboratory research use only. It is not intended for any animal or human therapeutic use, any human or animal consumption, or any diagnostic use.
Warranty

The product is provided 'AS IS' and the viability of ATCC® products is warranted for 30 days from the date of shipment, provided that the customer has stored and handled the product according to the information included on the product information sheet, website, and Certificate of Analysis. For living cultures, ATCC lists the media formulation and reagents that have been found to be effective for the product. While other unspecified media and reagents may also produce satisfactory results, a change in the ATCC and/or depositor-recommended protocols may affect the recovery, growth, and/or function of the product. If an alternative medium formulation or reagent is used, the ATCC warranty for viability is no longer valid.  Except as expressly set forth herein, no other warranties of any kind are provided, express or implied, including, but not limited to, any implied warranties of merchantability, fitness for a particular purpose, manufacture according to cGMP standards, typicality, safety, accuracy, and/or noninfringement.

Disclaimers

This product is intended for laboratory research use only. It is not intended for any animal or human therapeutic use, any human or animal consumption, or any diagnostic use. Any proposed commercial use is prohibited without a license from ATCC.

While ATCC uses reasonable efforts to include accurate and up-to-date information on this product sheet, ATCC makes no warranties or representations as to its accuracy. Citations from scientific literature and patents are provided for informational purposes only. ATCC does not warrant that such information has been confirmed to be accurate or complete and the customer bears the sole responsibility of confirming the accuracy and completeness of any such information.

This product is sent on the condition that the customer is responsible for and assumes all risk and responsibility in connection with the receipt, handling, storage, disposal, and use of the ATCC product including without limitation taking all appropriate safety and handling precautions to minimize health or environmental risk. As a condition of receiving the material, the customer agrees that any activity undertaken with the ATCC product and any progeny or modifications will be conducted in compliance with all applicable laws, regulations, and guidelines. This product is provided 'AS IS' with no representations or warranties whatsoever except as expressly set forth herein and in no event shall ATCC, its parents, subsidiaries, directors, officers, agents, employees, assigns, successors, and affiliates be liable for indirect, special, incidental, or consequential damages of any kind in connection with or arising out of the customer's use of the product. While reasonable effort is made to ensure authenticity and reliability of materials on deposit, ATCC is not liable for damages arising from the misidentification or misrepresentation of such materials.

Please see the material transfer agreement (MTA) for further details regarding the use of this product. The MTA is available at www.atcc.org.

Frequently Asked Questions

References

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