Liver Organoids | Organoids

Use cases

Ductal expansion organoids—not long-term 2D primary hepatocytes and not HCC cell-line spheroids.

Use	Fit	Readout	Notes
Adult liver ductal organoid expansion / disease modeling	Recommended	Expansion, genome stability, differentiation^[1]	Huch system supports long-term expansion and some genetic-disease mirroring.
Tox / metabolism pilots (after differentiation)	Contrast	Albumin, CYP, and related functional readouts	Expansion state is not a mature hepatocyte substitute.
HepG2 spheroids as “liver organoids”	Do not mix	Spheroid diameter	Cell-line spheroids are not adult ductal organoids.

In one line

Duct-enriched liver tissue → dome → Rspo/EGF/FGF10/HGF niche → brief Y-27632 → mechanical split → CK19/EpCAM → optional hepatocyte differentiation.

Protocol overview

Ethics + cold chain; resection or biopsy liver tissue
Trim fat/necrosis; collagenase ± Dispase; enrich ductal epithelium
Matrix domes; Huch-type medium (Rspo, EGF, FGF10, HGF, nicotinamide, A83-01, forskolin…)[[1]]
Brief Y-27632; score cystic ductal growth
Feed every 2–3 days; mechanical split by growth
CK19 / CK7 / EpCAM; mycoplasma; albumin usually low in expansion
Switch to differentiation conditions only when hepatocyte function is needed[[1]]
In vivo take uses transplant SOPs

Step 1 · Tissue digest

Log site (peripheral parenchyma favors duct enrichment).
Collagenase ± Dispase to clusters; filter and spin.
If bloody, brief RBC lysis (MC427); do not grind to pure single cells.

Steps 2–3 · Expansion niche

Start with a full Huch-type recipe^[1].
Mechanical split after cystic structures appear; log passage and matrix lot.
Quarantine immediately if contaminated or mycoplasma-positive.

Steps 4–5 · QC and differentiation

Expansion: CK19 / CK7 / EpCAM; albumin usually modest.
Differentiation: drop expansion keystones; add OSM, dexamethasone etc. per literature—separate dishes (see dedicated section).
In vivo confirmation uses transplant models—not this page.

Niche reagents (titrate)

Literature starting points—not on-site prescribed doses.

Component	Role	On-site
Advanced DMEM/F12 + B-27	Basal	Open →
R-spondin + EGF + FGF10 + HGF	Huch core^[1]	Open →
烟酰胺 / A83-01 / Forskolin	Common in human liver expansion	Open →
基底膜基质（BME）	Dome embedding	Open →
Y-27632 短期	Seeding window	Open →
红细胞裂解液	RBC lysis after bloody digests	Open →
TrypLE（温和传代备选）	Brief single-cell needs; prefer mechanical in expansion	Open →
OSM（分化期）	Hepatocyte-like differentiation keystone	Open →
地塞米松（分化期）	Pair with OSM etc.; off in expansion	Open →
VEGF-A（血管共培养扩展）	Off in default expansion/diff; endothelial co-culture separate	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 →

Expansion vs hepatocyte-like differentiation

Default here is ductal expansion. For hepatocyte function (albumin / CYP…), start separate dishes: withdraw expansion keystones (Rspo/EGF/FGF10/HGF…), then add OSM, dexamethasone, etc. per literature. Do not expand and differentiate in the same dish.

Scenario	Need CSS?	Plan
Long-term expansion, banking, genome edits	No	Huch expansion medium; do not maintain on OSM / Dex.
Hepatocyte-like function / tox pilots	Yes	Differentiation dish: OSM (MGF30014) + dexamethasone (MC429) titrated to literature; albumin Ab etc. as readout.
Need both expansion and functional readout	Yes	Split dishes: one bank in expansion, one switch to differentiation—do not mix media.

·In this section the needCss column means “switch to differentiation medium”, not serum CSS.
·Differentiation windows drift by lot—log OSM / Dex lots and days.
·Low albumin in expansion does not mean banking failed.

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
Forskolin	MCK1832	Adenylate cyclase; cAMP; common in ovarian recipes	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
Recombinant Human HGF	MGF30010	Human liver ductal organoid keystone	Protein	On request
Recombinant Human IGF-1	MGF30013	Seen in some epithelial / breast / liver recipes	Protein	On request
Recombinant Human Oncostatin M	MGF30014	Keystone for hepatocyte-like differentiation of liver organoids	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
CK19	Cytokeratin 19 Antibody	MCAB5312	Ductal-like epithelium; pancreas, some lung / ovarian / bladder / HNC
EpCAM	EPCAM Antibody	MCAB5377	Pan-epithelial; multi-tissue PDO
CK7	Cytokeratin 7 Antibody	MCAB5547	Cholangiocyte / urothelial / some tubular / esophageal / thyroid / mesothelial epithelium QC
Albumin	Albumin (MXB14128) Rabbit mAb	MDAB15418	Hepatocyte-like differentiation readout (often low in expansion)
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
N-2 Supplement (100×)	MC318	Neural / some epithelial recipes; use with or vs B-27 by system
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
ITS Supplement (100×)	MC311	Insulin–transferrin–selenium; some epithelial recipes
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
TrypLE Express Enzyme	MC426	Gentle PSC / organoid passaging alternative
RBC Lysis Buffer	MC427	RBC removal after bloody digests
Dexamethasone	MC429	Glucocorticoid for hepatocyte-like differentiation

Inquire for this list Culture supplements

References

[1] Long-term culture of genome-stable bipotent stem cells from adult human liver. Cell. 2015. PMID 25533785
[2] In vitro expansion of single Lgr5+ liver stem cells induced by Wnt-driven regeneration. Nature. 2013. PMID 24332895
[3] Human hepatic organoids for the analysis of human genetic diseases. JCI Insight. 2017. PMID 28218919

Human liver ductal organoid banking protocol