Head & Neck Cancer PDO | Organoids

Use cases

HNC PDO—not FaDu / Cal27 spheroids and not a full normal-mucosa expansion course.

Use	Fit	Readout	Notes
HNSCC / salivary resection or biopsy PDO biobank + chemo/RT	Recommended	Genomic fidelity and RT sensitivity vs clinical response^[1]	Millen / Driehuis biobank shows predictive potential especially in adjuvant RT settings.
Matched normal mucosa contrast	Contrast	Toxicity / selectivity	Keep normal and tumor dishes separate; do not mix drug plates.
FaDu / Cal27 spheroids as PDO	Do not mix	Spheroid diameter	Cell-line spheroids are not PDOs.

In one line

HNC clusters → dome → EGF/Rspo niche → brief Y-27632 → mechanical split → CK/p53 → chemo/RT drugs.

Protocol overview

Ethics + cold chain; log site (oral / oropharynx / larynx / hypopharynx / salivary) and HPV / smoking history
Trim necrosis; collagenase ± Dispase to clusters; brief RBC lysis (MC427) if bloody
Matrix domes; epithelial niche (EGF, FGF, Noggin, R-spondin ± Wnt, A83-01…)[[1]][[2]]
Brief Y-27632; score cystic vs solid growth
Feed every 2–3 days; mechanical split; bank SCC vs salivary separately
CK, EpCAM, p53; mycoplasma; match the primary
Drug / RT arms with vehicle; in vivo uses transplant SOPs—not this page

Step 1 · Tumor processing

Log anatomic site and HPV status; separate instruments.
Collagenase (MC412) ± Dispase (MC413) to clusters; brief RBC lysis (MC427) if bloody.
Do not grind to pure single cells.

Steps 2–3 · Niche and expansion

Start with a full epithelial recipe^[1]^[2]; bank SCC vs salivary separately.
If stroma overtakes, raise A83-01 or shorten digest.
Prefer mechanical splits; log passage and matrix lot.

Steps 4–5 · QC and chemo/RT

CK / EpCAM / p53; mycoplasma; match the primary.
Include vehicle on drug/RT arms; interpret vs clinical history^[1].
In vivo confirmation uses transplant SOPs—not this page.

Niche reagents (titrate)

Literature starting points—not on-site prescribed doses.

Component	Role	On-site
EGF + Noggin + R-spondin ± Wnt3a	HNC epithelial core^[1]^[2]	Open →
FGF2 / FGF10（按文献）	Common in epithelial niches	Open →
A83-01 / 烟酰胺	TGF-β block; human epithelial stability	Open →
Y-27632 短期	Seeding window	Open →
红细胞裂解液	Brief RBC lysis after bloody digests	Open →
超低吸附 / 圆底读板	Drug / RT contrast readouts	Open →
胶原酶 II 型	Digest tissue/tumor to epithelial clusters	Open →
Dispase II	Gentle dissociation with collagenase	Open →
类器官冻存液	Banking cryopreservation	Open →

Small molecules for this SOP

Deep-links to catalog SKUs; all prices on request. Growth factors (EGF/Noggin/R-spondin, etc.) are proteins—see that category.

Compound	SKU	Role	Price
Y-27632 2HCl	MCK1042	ROCK block; brief survival boost after seed/split	On request
SB 202190	MCK1279	p38 block; human epithelial organoid stability	On request
A83-01	MCK1825	ALK4/5/7; limit stromal overgrowth	On request
Nicotinamide	MCK1836	Common in human epithelial organoid media	On request

Inquire for this list All small molecules

Proteins / peptides for this SOP

Growth factors and gastrin deep-linked by SKU; all prices on request. Activity/reconstitution per lot COA.

Product	SKU	Role	Type	Price
Recombinant Human EGF	MGF30001	Core epithelial growth factor	Protein	On request
Recombinant Human Noggin	MGF30002	BMP antagonist; ENR / PDO niche; dual-SMAD neural induction	Protein	On request
Recombinant Human R-spondin-1	MGF30003	Wnt amplifier; long-term expansion	Protein	On request
Recombinant Human Wnt3a	MGF30004	Human colon / CRC / ductal / gastric / ovarian / liver Wnt	Protein	On request
Recombinant Human FGF2	MGF30006	Prostate niches; also common in brain / PSC induction	Protein	On request
Recombinant Human FGF10	MGF30007	Prostate / pancreas / breast / lung / gastric / ovarian / liver / endometrial / kidney niche	Protein	On request

Inquire for this list Proteins / enzymes / peptides

QC antibodies for this SOP

Common IHC/IF markers before banking or drugs; deep-linked catalog antibodies. Dilution/species per datasheet.

Marker	Product	SKU	Role
CK19	Cytokeratin 19 Antibody	MCAB5312	Ductal-like epithelium; pancreas, some lung / ovarian / bladder / HNC
EpCAM	EPCAM Antibody	MCAB5377	Pan-epithelial; multi-tissue PDO
E-Cadherin	E Cadherin Antibody	MCAB1155	Epithelial integrity / EMT contrast
p53	p53 Mouse mAb	B10301	Common in HGSOC / esophageal / HNC etc.; match to the primary

Inquire for this list Antibodies

Culture supplements for this SOP

Existing cell-culture SKUs (B-27 / N-2 / glutamine / antibiotics); titrate per insert and literature.

Product	SKU	Role
Advanced DMEM/F12 Medium	MC410	Serum-free organoid basal medium
B-27 Supplement (50×)	MC317	Serum-free basal supplement for epithelial / neural organoids
Stable glutamine (Ala-Gln, 200 mM)	MC301	Glutamine source for Advanced DMEM/F12 systems
Penicillin-Streptomycin (100×)	MC306	Early banking sterility; consider withdrawal after lines stabilize
Organoid Basement Membrane Extract (BME)	MC411	3D dome / sandwich embedding
Collagenase Type II	MC412	Digest tissue/tumor to epithelial clusters
Dispase II	MC413	Gentle dissociation with collagenase
Accutase Cell Detachment Solution	MC414	Optional gentle enzymatic passaging
Organoid Cryopreservation Medium	MC415	Banking cryopreservation
Mycoplasma Detection Kit	MC416	Pre-banking sterility QC
HEPES Buffer (1 M)	MC305	Organoid medium buffering; add to final concentration per recipe
Collagenase Type IV	MC421	Dissociation alternative; contrast with Type II
Organoid Primary Antimicrobial (Primocin-class)	MC423	Early primary / PDO / PSC banking contamination control
Ultra-Low Attachment 96-Well Plate	MC425	EB formation / spheroids / drug plates
U-Bottom 96-Well Plate	MC430	Spheroid settling / co-culture plates; complements ULA
RBC Lysis Buffer	MC427	RBC removal after bloody digests

Inquire for this list Culture supplements

Drugs ↔ spheroid / modeling / transplant roles

PDO drug assays stay in-matrix on this SOP. Cell-line spheroid / 2D viability lives on the spheroid hub and disease models; in vivo take and PDX use transplant models. Do not mix the three workflows.

Spheroid culture Disease models Transplant models

References

[1] Patient-derived head and neck cancer organoids allow treatment stratification and serve as a tool for biomarker validation and identification. Med. 2023. PMID 37178682
[2] Generation and Characterization of Patient-Derived Head and Neck, Oral, and Esophageal Cancer Organoids. Curr Protoc Stem Cell Biol. 2020. PMID 32294323

Head and neck cancer PDO banking protocol