Uveal Melanoma PDO | Organoids

Use cases

Uveal melanoma PDO—not a renamed cutaneous melanoma SOP and not 92.1 spheroids.

Use	Fit	Readout	Notes
Uveal melanoma PDO biobank + translational drugs	Recommended	Disease mirroring and resistance phenotypes^[1]^[2]	PDOs often mirror treatment resistance better than commercial lines.
Paired contrasts vs cutaneous melanoma	Contrast	Site-specific drug differences	Drivers and metastatic biology differ—bank separately.
92.1 / Mel270 spheroids as PDO	Do not mix	Spheroid diameter	Cell-line spheroids are not PDOs.

In one line

Uveal-tumor clusters → dome → EGF/FGF2 niche → brief Y-27632 → passage → SOX10/Melan-A → drugs.

Protocol overview

Ethics + cold chain; log intraocular site and GNAQ / GNA11 / BAP1 status
Trim necrosis; gentle digest to clusters; brief RBC lysis (MC427) if bloody
Matrix domes; EGF / FGF2 ± Noggin / Rspo[[1]][[2]]
Brief Y-27632; score solid growth and pigment
Feed every 2–3 days; mechanical split; bank separately from cutaneous melanoma
SOX10, MiTF, Melan-A; mycoplasma; match the primary
Drugs with vehicle; interpret vs commercial-line resistance gaps[[2]]

Step 1 · Tissue and digest

Log intraocular site and molecular class; separate instruments from cutaneous specimens.
Collagenase (MC412) ± Dispase (MC413) to clusters.
Do not grind to pure single cells.

Steps 2–3 · Niche and expansion

Start with EGF / FGF2; add Noggin/Rspo only if literature-backed^[1]^[2].
Keep media lots separate from cutaneous melanoma.
Mechanical splits; log pigment and passage.

Steps 4–5 · QC and drugs

SOX10 / MiTF / Melan-A; mycoplasma; match the primary.
Include vehicle; interpret vs commercial-line gaps^[2].
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 + FGF2	Uveal melanoma core^[1]^[2]	Open →
Noggin / R-spondin（可选）	Titrate by literature	Open →
Y-27632 短期	Seeding window	Open →
超低吸附 / 圆底读板	Drug 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
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 VEGF-A	MGF30015	Vascular / endothelial co-culture extension; usually off in default epithelial PDO media	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
p53	p53 Mouse mAb	B10301	Common in HGSOC / esophageal / HNC etc.; match to the primary
SOX10	SOX10 Antibody	MCAB5826	Melanoma / neural-crest lineage QC
MiTF	MiTF Antibody	MCAB6661	Melanocytic differentiation TF QC
Melan-A	MelanA Antibody	MCAB5604	Melanocytic differentiation marker; cutaneous / uveal melanoma QC

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] Novel Uveal Melanoma Patient-Derived Organoid Models Recapitulate Human Disease to Support Translational Research. Invest Ophthalmol Vis Sci. 2024. PMID 39601636
[2] Novel uveal melanoma patient-derived cell line and organoid models show increased treatment resistance compared with commercially available models. Melanoma Res. 2026. PMID 42108204

Uveal melanoma PDO banking protocol