Kidney Organoids | Organoids

Use cases

Kidney tubuloids ± RCC PDO—not a full iPSC multi-lineage kidney course and not 786-O spheroids.

Use	Fit	Readout	Notes
Adult kidney / urine tubuloid disease modeling	Recommended	Tubular architecture, tubular markers, infection / genetics^[1]	Schutgens tubuloids expand long-term for personalized modeling.
RCC resection PDO (separate dishes)	Contrast	Tumor markers / drugs	Bank strictly apart from normal tubuloids; titrate medium to literature.
786-O / A498 spheroids as kidney organoids	Do not mix	Spheroid diameter	Cell-line spheroids are not tubuloids / PDOs.

In one line

Kidney tissue / urine epithelium → dome → EGF/Rspo niche → brief Y-27632 → mechanical split → tubular markers → optional RCC dishes.

Protocol overview

Ethics + cold chain; log site (cortex / medulla / urine) and tumor vs normal
Normal kidney: enzyme to clusters; urine: spin-enrich epithelium; brief RBC lysis (MC427) if bloody
Matrix domes; tubuloid medium (EGF, FGF, R-spondin, A83-01…)[[1]]
Brief Y-27632; score tubular / cystic structures
Feed every 2–3 days; mechanical split; log passage and matrix lot
Tubular markers (CK7 / EpCAM etc. by study); mycoplasma
RCC / tumor PDO: separate dishes and recipes—do not mix with normal tubuloids
iPSC multi-lineage kidney induction is a different study; in vivo uses transplant SOPs

Step 1 · Tissue and digest

Separate instruments for normal vs tumor; log cortex / medulla.
Enzyme to clusters; spin-enrich urine specimens; brief RBC lysis (MC427) if bloody.
Do not grind to pure single cells.

Steps 2–3 · Tubuloid expansion

Start with a Schutgens-class recipe^[1]; score tubular morphology.
Prefer mechanical splits; log passage.
Open separate dishes for RCC—never mix with normal banks.

Steps 4–5 · QC and boundaries

CK7 / EpCAM tubular / epithelial readouts; mycoplasma.
This page is not iPSC multi-lineage kidney induction (Takasato etc. are separate).
In vivo orthotopic renal take uses 786-O / A498 transplant SOPs.

Niche reagents (titrate)

Literature starting points—not on-site prescribed doses.

Component	Role	On-site
EGF + R-spondin + FGF（按文献）	Tubuloid core^[1]	Open →
A83-01	TGF-β / stroma	Open →
烟酰胺（常见）	Human epithelial stability	Open →
Y-27632 短期	Seeding window	Open →
红细胞裂解液	Brief RBC lysis if bloody	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 R-spondin-1	MGF30003	Wnt amplifier; long-term expansion	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
EpCAM	EPCAM Antibody	MCAB5377	Pan-epithelial; multi-tissue PDO
PAX8	PAX8 Antibody	MCAB5911	Ovarian / Müllerian / endometrial / renal epithelial QC
CK7	Cytokeratin 7 Antibody	MCAB5547	Cholangiocyte / urothelial / some tubular / esophageal / thyroid / mesothelial epithelium 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.

Suggested cell-line spheroid contrast

Quote: RCC spheroid starter →

Spheroid culture Disease models Transplant models

References

[1] Tubuloids derived from human adult kidney and urine for personalized disease modeling. Nat Biotechnol. 2019. PMID 30833775
[2] Kidney organoids from human iPS cells contain multiple lineages and model human nephrogenesis. Nature. 2015. PMID 26466099

Human kidney tubuloid banking protocol