Cell culture · 3D & organoid workflows
Organoids are 3D cultures that approximate organ structure and function. Use tissue cards, a core reagent map, and five general protocol outlines—then jump to media, supplements, small molecules, growth factors, and modeling resources.
Separate 3D formats from in vivo models before picking plates and SOPs—don’t call cell-line spheroids or CDX a PDO.
Usually simple 3D aggregates of cell lines—good for drug throughput, not organ patterning. Use U-bottom (MC430) or ULA (MC425) plates.
Spheroid hub (wired to cell lines) →Adult stem cells or patient tissue self-organize in niche medium + matrix; our SOPs default to dome embedding. Organ-on-chip is a separate project—not prescribed here.
CDX/PDX and orthotopic grafts answer in vivo take and PK questions; a Matrigel plug ≠ an organoid dome. For in vivo follow-up, use disease models / transplant models.
Capture these readouts for every PDO / organoid bank before drugs or editing.
| Item | Suggested practice |
|---|---|
| Morphology & growth | Photos pre/post split; log diameter, cystic/solid, core necrosis |
| Passage success | Fix split ratio and digest time; quarantine failed lots |
| Lineage / tumor markers | SOP QC Abs (CK, EpCAM, tissue TFs…) vs the primary |
| Mycoplasma / sterility | Test at banking (MC416); discard positives; early MC423 optional |
| STR / identity (PDO) | When available, match primary DNA to catch cross-contamination |
| Matrix & factor lots | Log BME (MC411), B-27, growth-factor and small-molecule lots |
Lock tissue and niche first. Cards marked “dedicated protocol” open a step-by-step SOP; others link to pathways or modeling hubs.
Wnt / R-spondin / Noggin / EGF; adult-crypt step-by-step protocol
Human-colon niche; APC-mutant lines often Wnt-independent; drug biobank
NRG1 + EGF/FGF; ER/HER2 subtype–aware; step-by-step banking
DHT + Wnt/Noggin/EGF + A83-01; step-by-step banking protocol
Wnt/FGF10 ductal niche; Seino subtype; step-by-step banking
EGF/FGF2; pNET/GEP-NET; step-by-step banking
FGF7/10 + EGF; airway-like niche; step-by-step banking
Wnt/FGF10/gastrin; gastric niche; step-by-step banking
Wnt/EGF ± gastrin; EAC/ESCC; Barrett caution
EGF/FGF/Rspo; SCC / salivary; step-by-step banking
Rspo/FGF10 ± Wnt; PAX8 QC; step-by-step banking
EGF/FGF/Noggin/Rspo; urothelial niche; step-by-step banking
EGF/FGF/Rspo; tubuloid; step-by-step protocol
EGF/Rspo/FGF10 ±E2; endometrial niche; step-by-step banking
EGF/FGF/Rspo; HPV-aware; step-by-step banking
EGF/FGF/Rspo; PTC niche; step-by-step banking
EGF/FGF/Rspo; ALI stratification; step-by-step protocol
EGF/FGF2; melanocytic tumor; step-by-step banking
EGF/FGF2; uveal melanoma; step-by-step banking
HGF + FGF10 + Rspo; ductal expansion; step-by-step protocol
HGF+FGF10+Rspo; CCA; step-by-step banking
EGF/FGF/Rspo; gallbladder epithelium; step-by-step banking
EGF/FGF2 ±HGF; mesothelioma; step-by-step banking
EGF/FGF2; matrix-sensitive; step-by-step banking
EGF/FGF2; soft-tissue sarcoma; step-by-step banking
EGF/FGF2 + B-27; GBO; step-by-step banking
B-27 / N-2; dual-SMAD & CHIR windows; step-by-step protocol
Cross-compare live SOPs; Usual / Optional / — is a lookup only—not a dosing recipe.
| Factor | Gut | CRC | Breast | Prostate | PDAC | pNET | Lung | Gastric | Eso. | HNC | Ovarian | Bladder | Kidney | Endo. | Cervix | Thyroid | Skin | Melanoma | Uveal | Liver | CCA | GB | Meso | OS | STS | GBM | Brain |
|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
| EGF | Usual | Usual | Usual | Usual | Usual | Usual | Usual | Usual | Usual | Usual | Usual | Usual | Usual | Usual | Usual | Usual | Usual | Usual | Usual | Usual | Usual | Usual | Usual | Usual | Usual | Usual | — |
| Noggin | Usual | Usual | Usual | Usual | Usual | Usual | Usual | Usual | Usual | Usual | Usual | Usual | Optional | Usual | Usual | Usual | Usual | Optional | Optional | Optional | Usual | Usual | Optional | — | — | — | Usual |
| R-spondin-1 | Usual | Usual | Usual | Usual | Usual | Usual | Usual | Usual | Usual | Usual | Usual | Usual | Usual | Usual | Usual | Usual | Usual | Optional | Optional | Usual | Usual | Usual | Optional | — | — | — | — |
| Wnt3aOften for human colon/stomach; gallbladder Wnt as separate stim/inhib arms | Optional | Optional | — | Optional | Usual | Optional | Optional | Usual | Optional | Optional | Optional | Optional | — | — | Optional | — | Optional | Optional | Optional | Optional | Optional | Optional | Optional | — | — | — | — |
| NRG1 | — | — | Usual | — | — | — | Optional | — | — | — | — | — | — | — | — | — | — | — | — | — | — | — | — | — | — | — | — |
| HGF | — | — | — | — | — | — | — | — | — | — | — | — | — | — | — | — | — | — | — | Usual | Usual | Optional | Optional | — | — | — | — |
| FGF2 | — | — | — | Usual | — | Usual | — | — | Optional | Optional | — | Optional | Optional | — | Optional | Optional | Optional | Usual | Usual | — | — | — | Usual | Usual | Usual | Usual | Optional |
| FGF7 / FGF10 | — | — | Usual | Usual | Usual | — | Usual | Usual | Usual | Usual | Usual | Usual | Usual | Usual | Usual | Usual | Usual | — | — | Usual | Usual | Usual | Optional | — | — | — | — |
| Activin A / BMP4PSC lineage-window contrast | — | — | — | — | — | — | — | — | — | — | — | — | — | — | — | — | Optional | — | — | — | — | — | — | — | — | — | Optional |
| DHT | — | — | — | Usual | — | — | — | — | — | — | — | — | — | — | — | — | — | — | — | — | — | — | — | — | — | — | — |
| E2ER+ breast / endometrium by subtype | — | — | Optional | — | — | — | — | — | — | — | — | — | — | Optional | — | — | — | — | — | — | — | — | — | — | — | — | — |
| Gastrin | — | — | — | — | Usual | — | — | Usual | Optional | — | — | — | — | — | — | — | — | — | — | — | — | — | — | — | — | — | — |
| Forskolin | — | — | — | — | — | — | — | — | — | — | Usual | — | — | — | — | — | Optional | — | — | Usual | Usual | Usual | — | — | — | — | — |
| Dual-SMAD (SB431542 / LDN) | — | — | — | — | — | — | — | — | — | — | — | — | — | — | — | — | — | — | — | — | — | — | — | — | — | — | Optional |
| CHIR-99021pNET drug contrast arm—not default expansion | — | — | — | — | — | Optional | — | — | — | — | — | — | — | — | — | — | — | — | — | — | — | — | — | — | — | — | Optional |
| A83-01 | Optional | Usual | Optional | Usual | Usual | Optional | Usual | Usual | Usual | Usual | Usual | Usual | Usual | Usual | Usual | Usual | Usual | Optional | Optional | Usual | Usual | Usual | Optional | Optional | Optional | — | — |
| Y-27632 (brief) | Usual | Usual | Usual | Usual | Usual | Usual | Usual | Usual | Usual | Usual | Usual | Usual | Usual | Usual | Usual | Usual | Usual | Usual | Usual | Usual | Usual | Usual | Usual | Usual | Usual | Usual | Usual |
| OSMHepatocyte-like differentiation; off in expansion | — | — | — | — | — | — | — | — | — | — | — | — | — | — | — | — | — | — | — | Optional | — | — | — | — | — | — | — |
| DexamethasoneOften with OSM etc. | — | — | — | — | — | — | — | — | — | — | — | — | — | — | — | — | — | — | — | Optional | — | — | — | — | — | — | — |
| VEGF-AVascular co-culture extension; off in default epithelial media | — | — | — | — | — | — | Optional | — | — | — | — | — | — | — | — | — | Optional | Optional | Optional | Optional | — | — | Optional | Optional | Optional | Optional | Optional |
| XAV939 / IWP-2Wnt-inhibition contrast arms—not default expansion | Optional | Optional | — | — | — | — | — | — | Optional | Optional | — | — | — | — | Optional | — | — | — | — | — | — | Optional | — | — | — | — | — |
| ATRAAirway–alveolar / lineage / epidermal windows | — | — | — | — | — | — | Optional | — | — | — | — | — | — | — | — | — | Optional | — | — | — | — | — | — | — | — | — | Optional |
| ProgesteroneHormone arms with E2 | — | — | Optional | — | — | — | — | — | — | — | — | — | — | Optional | — | — | — | — | — | — | — | — | — | — | — | — | — |
Common modules and on-site entry points—titrate doses/timing to tissue literature; not a prescription.
| Reagent | Role | On-site |
|---|---|---|
| B-27 / N-2 | Serum-free neural/epithelial supplements for brain and multi-lineage organoids | B-27 SKU MC317 → |
| ITS / L-Gln / NEAA / 双抗 | Fine-tune nutrition and selection pressure; log lots and final concentrations | Supplements → |
| Y-27632 | ROCK inhibition; short-term boost after single-cell seeding | Cytoskeleton pathway → |
| CHIR99021 | GSK-3 block → Wnt activation; stemness and lineage induction | Wnt pathway → |
| A83-01 / SB431542 | ALK4/5/7; suppress TGF-β / Activin / Nodal | TGF-β/Smad pathway → |
| EGF / FGF / Noggin / R-spondin / Wnt | Growth factors set identity and proliferation; prefer activity-validated lots | Proteins / enzymes / peptides → |
| 解离酶 / 冻存液 / 支原体检测 | Gentle passaging, banking, and sterility QC | Collagenase MC412 → |
| 基底膜基质（BME） | 3D dome / sandwich embedding | BME MC411 → |
| 超低吸附 / 圆底板 | ULA (MC425) anti-attachment; U-bottom (MC430) for settling | ULA MC425 · U-bottom MC430 → |
Core SOP compounds plus common extensions; all prices on request. Growth factors are under proteins.
| Compound | SKU | Role | Tier |
|---|---|---|---|
| Y-27632 2HCl | MCK1042 | ROCK block; brief survival boost after seed/split | Existing |
| CHIR-99021 | MCK1040 | GSK-3 block → Wnt on; stemness/lineage induction | Existing |
| SB 431542 | MCK1038 | ALK4/5/7; dual-SMAD / TGF-β block | Existing |
| SB 202190 | MCK1279 | p38 block; human epithelial organoid stability | Existing |
| DAPT | MCK1021 | γ-secretase / Notch; differentiation windows | Existing |
| PD0325901 | MCK1028 | MEK; PSC dual inhibition / lineage switches | Existing |
| A83-01 | MCK1825 | ALK4/5/7; limit stromal overgrowth | SOP core |
| RepSox | MCK1826 | ALK5; TGF-β block alternative (vs A83-01) | Common extend |
| Thiazovivin | MCK1827 | ROCK; single-cell survival alternative (vs Y-27632) | Common extend |
| (-)-Blebbistatin | MCK1828 | Myosin II; reduce hyper-contraction | Common extend |
| LDN193189 | MCK1829 | BMP; dual-SMAD / lineage induction | Common extend |
| IWP-2 | MCK1830 | Porcupine; block Wnt secretion; CRC Wnt-withdraw contrast | Common extend |
| XAV939 | MCK1831 | Tankyrase; suppress Wnt; CRC / gut contrast arms | Common extend |
| Forskolin | MCK1832 | Adenylate cyclase; cAMP; common in ovarian recipes | Common extend |
| Prostaglandin E2 | MCK1833 | Common in prostate organoid recipes | SOP core |
| 5α-Dihydrotestosterone | MCK1834 | Androgen for AR+ prostate organoids | SOP core |
| 17β-Estradiol | MCK1835 | Hormone arm for ER+ breast organoids | SOP core |
| Progesterone | MCK1838 | Breast / endometrial hormone arms; with E2 on separate dishes | SOP core |
| All-trans Retinoic Acid (ATRA) | MCK1837 | Lung airway–alveolar bias windows; usually off in default expansion | Common extend |
| Nicotinamide | MCK1836 | Common in human epithelial organoid media | SOP core |
Price: on request · Browse small molecules
EGF, Noggin, R-spondin, Wnt3a, NRG1, FGFs, and gastrin; all prices on request. Activity 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 NRG1 | MGF30005 | Sachs breast keystone; optional in some lung recipes | 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 |
| Recombinant Human FGF7 | MGF30008 | Common in breast and airway epithelial recipes | Protein | On request |
| Human Gastrin-I | MGF30009 | Pancreatic ductal / gastric PDO peptide | Peptide | On request |
| Recombinant Human HGF | MGF30010 | Human liver ductal organoid keystone | Protein | On request |
| Recombinant Human Activin A | MGF30011 | PSC / brain induction windows (dual-SMAD lineage contrast) | Protein | On request |
| Recombinant Human BMP4 | MGF30012 | BMP axis; lineage induction / Noggin·LDN contrast | 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 |
Price: on request · Browse proteins / enzymes / peptides
Existing cell-culture SKUs (B-27, N-2, glutamine, antibiotics, ITS) deep-linked on SOP shopping lists.
| 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 |
| N-Acetyl-L-cysteine (NAC) | MC417 | Antioxidant in prostate and related recipes |
| Hydrocortisone | MC418 | Additive in breast / airway epithelial recipes |
| Charcoal-Stripped FBS (CSS) | MC419 | Serum + hormone manipulation only; not for default serum-free banking |
| HEPES Buffer (1 M) | MC305 | Organoid medium buffering; add to final concentration per recipe |
| Collagenase Type IV | MC421 | Dissociation alternative; contrast with Type II |
| Crypt-Release EDTA Working Solution | MC422 | Adult intestinal crypt EDTA release |
| Organoid Primary Antimicrobial (Primocin-class) | MC423 | Early primary / PDO / PSC banking contamination control |
| Neurobasal Medium | MC424 | Brain / neural / GBM organoid basal |
| 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 |
| Keratinocyte Serum-Free Medium (KSFM) | MC428 | Prostate CSS-style recipes; optional contrast for skin / epidermis |
| Dexamethasone | MC429 | Glucocorticoid for hepatocyte-like differentiation |
| Collagen Type I (rat tail / bovine) | MC431 | Osteosarcoma / soft-tissue sarcoma matrix contrast; lock lots vs BME |
Reference outlines—optimize with inserts, tissue type, and pilots. Tissue-specific SOPs are listed below.
Dedicated step-by-step protocols
Tip: Log matrix lot and gel:medium volume ratio for reproducibility. SKU: BME MC411.
Tip: Chronic ROCK inhibition can bias lineage—do not leave it on by default. SKUs: Y-27632 MCK1042; alt Thiazovivin MCK1827.
Tip: Over-digestion collapses 3D architecture and stemness—err on the short side. Enzymatic options: Accutase MC414 / TrypLE MC426.
Tip: Treat as a separate book from 2D line SOPs—do not copy adherent DMSO habits unverified. SKU: organoid cryo medium MC415.
Tip: Antibiotics are not a substitute for aseptic technique and scheduled testing. SKUs: mycoplasma MC416; antimicrobial MC423; collagenase MC412 / Dispase MC413.
Matrix lots, edge effects, cryo, and contamination—conversion FAQs with SKU deep links.
Lock SKU and lot (BME MC411), log gel:cell ratios; run a small parallel pilot before switching the whole bank.
Prefer inner wells and limit evaporation (per lab SOPs); for drug plates pick ULA (MC425) or U-bottom (MC430) by goal and lock the layout.
Use organoid cryo medium (MC415); validate matrix removal vs micro-droplets; brief day-1 Y-27632 (MCK1042)—do not copy 2D DMSO habits.
Add Primocin-class (MC423) or P/S (MC306) early; quarantine contaminated lots; mycoplasma-test with MC416 and discard positives—do not rescue-mix.
Continue by reagent and consumable category pages.
Note: This is a workflow hub, not a substitute for product inserts. BME (MC411), dissociation enzymes, cryo medium, and CSS (MC419) are listed as inquiry SKUs; contact sales for other specialty reagents.