In Vitro 2D/3D Models

General Information

For each type of in vitro model:

The origin of material used to generate a specific model is listed in the PDMR database with the model growth information.
Complete human pathogen assessment has been performed as described in SOP50104. The PDMR does not distribute models that are positive for human immunodeficiency virus, Hepatitis B, or Hepatitis C.
Human origin has been confirmed.
STR profiles validated matching patient material and/or other models derived from the same patient material.
Model-specific Certificate of Analysis is provided with distributed material.
PDMR SOPs should be used to ensure culture success — these models should not be treated like traditional in vitro cultures (e.g., HeLa, MCF7).

For PDOrg and PDC models:

Derived xenografts are confirmed by histopathology to match the patient diagnosis or pathology of patient originator material.
Whole exome sequence and RNASeq data are available in the PDMR.

PDOrg models

Guaranteed for at least 10 passages (following PDMR SOPs)

PDOrg Model Generation

The PDMR generates PDOrg in vitro models from patient material and then PDXs, in that order of priority to try to maximize model heterogeneity.

PDOrg cultures use specialized defined media with growth supported in a basement membrane dome. The PDMR has noted 3 different morphological patterns of growth among PDOrgs: organoids, loose aggregate clusters, and single cell/small clusters.

PDOrg-Specific Quality Control

Verified to be 99.9% human tumor culture, non-clonal (multiple cell types or non-cloned lineages), 99.9% fibroblast free (FACs analysis).
Proven to grow as a cell-line derived xenograft (CLX) in NSG host mice. CLXs are characterized by histopathology for human tumor/diagnosis sub-type confirmation but are not further analyzed.
No out-growth of fibroblasts has been observed for at least 10 passages.
May or may not be able to form spheroids in a fully-defined, serum- and feeder-free medium

PDC models

Guaranteed for at least 20 passages (following PDMR SOPs)

PDC Model Generation

The PDMR generates PDC in vitro models from patient material, PDX, and patient/PDX-derived organoids (PDOrg) in that order of priority to try to maximize model heterogeneity.

PDC cultures use defined media and grow with a variety of growth characteristics.

PDC-Specific Quality Control

Verified to be 99.9% human tumor culture, non-clonal (multiple cell types or non-cloned lineages), 99.9% fibroblast free (FACs analysis).
Proven to grow as a cell-line derived xenograft (CLX) in NSG host mice. CLXs are characterized by histopathology for human tumor/diagnosis sub-type confirmation but are not further analyzed.
No out-growth of fibroblasts has been observed for at least 20 passages.
Tested for growth as spheroids in fully-defined, serum- and feeder-free medium and tested for clonal growth in soft agar.

CAF models

Guaranteed for at least 3 passages (following PDMR SOPs)
Non-tumorigenic in mice

CAF Model Generation

CAFs are primarily generated from patient material, though occasionally they can be recovered from a P0/P1 PDX.

CAF cultures use defined Media and are grown on Matrigel-coated plates.

CAF-Specific Quality Control

Verified to be 99.9% human fibroblast culture and 99.9% human tumor cell free by FACS analysis.
These cultures have been proven to be non-tumorigenic following subcutaneous implantation into NSG mice and have been characterized as fibroblasts by at least one of the following methods: immunohistochemistry, qRT-PCR, or FACs.
It is important to note these are non-transformed cells. CAFs have a finite lifespan in vitro.
CAFs are guaranteed for experimental use for up to 3 passages when maintained on Matrigel-coated surface in the recommended defined media.
Additional population doublings and subcultures are possible, but overall fitness of culture may deteriorate with subsequent passages.
Not sequenced for the Repository, though they may be used as a germline surrogate for the model.