Pancreatic Cancer PDO | Organoids

Use cases

PDAC PDO—not islet organoids and not non-tumor ductal expansion (contrast only; separate dishes).

Use	Fit	Readout	Notes
PDAC PDO drug profiling	Recommended	Viability and chemo response^[3]	Tiriac et al. showed organoids can stratify common chemo responses.
Normal pancreatic duct organoids	Contrast	Cystic expansion^[1]	Usually more Wnt-dependent; keep separate from tumor dishes.
Islet / β-cell differentiation	Do not mix	Insulin secretion	Endocrine specification is a different PSC/adult recipe.

In one line

Desmoplastic digest → dome → Wnt/FGF10 ductal medium → drop Wnt by Seino subtype → CK19 + mycoplasma → chemo.

Protocol overview

Ethics + cold chain; process EUS/resection tissue promptly
Desmoplastic tumors: longer collagenase, keep epithelial clusters[[1]]
Domes; ductal medium with Wnt3a, R-spondin, EGF, FGF10, Noggin, gastrin[[1]][[2]]
Brief Y-27632; A83-01 to limit TGF-β-driven fibroblasts
After stable growth, subtype Wnt dependence—some lines drop Wnt[[2]]
Cystic/budding morphology + CK19; mycoplasma
Gemcitabine / FOLFIRINOX arms as relevant[[3]]
Islet/endocrine induction is not this SOP

Step 1 · Stromal digest

PDAC is fibrotic—digests often run longer than gut/prostate, but still aim for clusters not single cells^[1].
Spin off RBC/debris; brief RBC lysis (MC427) if bloody; optional brief plating to shed fibroblasts without losing epithelial clusters.

Steps 2–3 · Ductal niche and Wnt subtype

Bank in full Wnt/FGF10 medium^[1].
Seino subtypes: some advanced tumors lose niche-factor dependence—trial Wnt withdrawal^[2].
If growth stalls, restore Wnt before discarding as contamination.

Steps 4–5 · QC and drugs

CK19 ductal phenotype; widespread islet-hormone positivity is out of scope unless that is the study.
Dose after stable passage; log matrix lot^[3].

Niche reagents (titrate)

Literature starting points—not on-site prescribed doses.

Component	Role	On-site
Wnt3a + R-spondin + EGF + FGF10 + Noggin	Ductal core^[1]	Open →
胃泌素、烟酰胺、PGE2（按文献）	Common in human pancreas recipes^[2]	Open →
A83-01	TGF-β / stroma^[2]	Open →
Y-27632 短期	Seeding survival	Open →
红细胞裂解液	Brief RBC lysis after bloody digests	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 FGF10	MGF30007	Prostate / pancreas / breast / lung / gastric / ovarian / liver / endometrial / kidney niche	Protein	On request
Human Gastrin-I	MGF30009	Pancreatic ductal / gastric PDO peptide	Peptide	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
Ki-67	Ki67 Rabbit mAb	B10315	Proliferation index; pre/post-drug contrast

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.

Suggested cell-line spheroid contrast

Quote: Pancreas spheroid starter →

Spheroid culture Disease models Transplant models

References

[1] Organoid models of human and mouse ductal pancreatic cancer. Cell. 2015. PMID 25557080
[2] Human pancreatic tumor organoids reveal loss of stem cell niche factor dependence during disease progression. Cell Stem Cell. 2018. PMID 29249464
[3] Organoid profiling identifies common responders to chemotherapy in pancreatic cancer. Cancer Discov. 2018. PMID 29853643

Pancreatic cancer PDO banking protocol