HCM-SANG-0272-C20
PDM-49 ™
PDM-49 ™
organoid
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.
Basic research, compound screening
ICD-10-CM code: C20 malignant neoplasm of rectum; primary adenocarcinoma
See associated clinical data for patient profile information, if available.
https://portal.gdc.cancer.gov/
https://hcmi-searchable-catalog.nci.nih.gov/model/HCM-SANG-0272-C20
To prepare the complete medium for this organoid model, please refer to the Organoid Media Formulation #4.
ATCC offers the recombinant proteins, small molecules, and other supplements to make this complete medium; Organoid Growth Kit 1D (ATCC ACS-7103) provides these supplements in a convenient, pre-portioned, ready-to-reconstitute format that does not require aliquoting or storage once prepared.
Seeding density: We recommend seeding this model at 0.25 - 1 x 106/ viable cells in 100 µL of ECM per well of a 6-well plate.
ECM: We recommend culturing this model in ATCC Cell Basement Membrane (ATCC ACS-3035) or Corning Matrigel. Include 10 µM ROCK Inhibitor Y-27632 (ATCC ACS-3030) in medium for the first 2-3 days following subculture.
For a brief overview of the thawing procedure see our quickstart guide Thawing Cryopreserved Human Organoids.
Initiating culture from frozen vials: For a brief overview of the thawing procedure see our quickstart guide Thawing Cryopreserved Human Organoids.
Seeding density: 0.25 - 1 x 106/ viable cells in 100 µL of ECM per well of a 6-well plate. Alternatively, split at 1:2-1:4 every 7-10 days. For example, collect organoids from 100 µL of extracellular matrix (ECM) from a single well of a 6-well plate and re-seed into 2-4 wells of a 6-well plate in 100 µL ECM per well.
Media renewal: Perform a complete medium change every 2-3 days. Include 10 µM ROCK Inhibitor Y-27632 (ATCC ACS-3030) in medium for the first 2-3 days following subculture.
For a brief overview of the subculture and expansion of organoids see our quickstart guide Subculture and Expansion of Human Organoids Protocol.
For more details on the handling and culture of organoids see our methods paper in Current Protocols in Cell Biology.
We recommend cryopreserving this model in ATCC Stem Cell Freezing Media (ATCC ACS-3020)
For a brief overview of the cryopreservation procedure for organoids see our quickstart guide Organoid Cryopreservation Protocol.
For more details on the handling and culture of organoids see our methods paper in Current Protocols in Cell Biology.
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