A complete strategy for building a colitis model

Inflammatory bowel disease (IBD) is a chronic inflammatory bowel disease that mainly involves the digestive system, among which Crohn's disease (CD) and ulcerative colitis (UC) are the main types. DSS (dextran sulfate sodium salt)-induced colitis is the most widely used research model of inflammatory bowel disease (IBD) in the world. Its core advantages are that it highly simulates human ulcerative colitis (UC) and accurately replicates clinicopathological features (colon shortening, crypt injury, goblet cell reduction, bloody stool), easy operation and good repeatability. In vivo experiments, animals were administered through drinking water (3-5% DSS, 5-7 days) without surgery or genetic modification. In vitro experiments, fibroblasts were isolated from colon surgical samples of UC patients, and then reprogrammed into iPSCs and differentiated into ulcerative colitis organoids iHUCOs.

 

1. Ulcerative colitis (UC) model in vivo

 

1.1 This model destroys the intestinal barrier and induces inflammation through DSS, and combined with IGF2 intervention to verify its protective effect. It is suitable for studying the mechanism and drug screening of ulcerative colitis [1].

 

(1) Experimental Animal

Wild-type C57BL/6J mice

 

(2) UC Modeling method

① The mice were kept in a room with controllable light (12-hour light/dark cycle) and temperature (22 ± 0. 5 ℃), eating and drinking freely;

② After a 1-week adaptation period, the mice drank free aqueous solution containing 5% (w/v) DSS (abs9192) for 7 days to establish a colitis model, and the control group drank sterile water.

 

(3) Evaluation Index

Colon shortening + weight loss + DAI score ≥ 3;

Histological verification: inflammatory infiltration, crypt injury, goblet cell reduction;

Molecular markers: increased MPO activity and upregulated pro-inflammatory factors (IL-1β/TNF-α/IL-6);

Accompanying phenomena: down-regulation of IGF2 expression, destruction of intestinal barrier, activation of cGAS-Sting pathway.

 

 

1.2 This model comprehensively evaluates UC and its neuropsychiatric complications through multi-dimensional indicators (intestinal pathology, behavior, molecular mechanism), and is suitable for studying intestinal-brain interactions and potential therapeutic targets [2].

 

(1) Experimental Animal

8-week-old male C57BL/6J mice

 

(2) UC Modeling method

① The mice were kept in a room with controllable light (12-hour light/dark cycle) and temperature (22 ± 2 ℃), and they were free to eat and drink.

② After a 1-week adaptation period, mice were free to drink aqueous solution containing 4% (w/v) DSS (abs9192) for 7 days to establish a colitis model, and the control group was set as follows:

Control group (Ctrl + NS): drink sterile water and inject saline (NS) intraperitoneally.

Model group (DSS + NS): 4% DSS water was drunk and NS was injected intraperitoneally.

Treatment group (DSS + Bet): drinking 4% DSS water and intraperitoneal injection of betaine (abs47001024, 600 mg/kg/day).

 

(3) Evaluation Index

Pathological changes of the colon: increased inflammation score, decreased goblet cells, shortened colon.

Behavioral abnormalities: FST/TST rest time prolonged, LDBT/OFT exploration behavior decreased.

Molecular Validation: Colonic M1 macrophage markers were upregulated and M2 was downregulated. Hippocampal DNA damage, mitochondrial dysfunction and activation of cGAS-STING pathway.

 

 

2. In vitro human ulcerative colitis organoids (iHUCOs) model [3]

 

2.1 Cell Source

 

Colonic fibroblasts were isolated from surgical specimens from patients with ulcerative colitis (UC).

 

2.2 UC Modeling method

 

(1) Reprogramming to iPSCs

Fibroblasts from UC patients were transformed into induced pluripotent stem cells (iPSCs) using the CytoTune-IPS reprogramming kit.

 

(2) Differentiation into intestinal organoids

Endoderm differentiation: iPSCs differentiated into stereotyped endoderm (DE) in medium containing Activin A (abs05801) for 3 days.

Intestinal spheroid formation: DE differentiated into intestinal spheroids (SPH) in medium containing FGF4 (abs06269) and CHIR99021 (abs810052) for 4 days.

Organoid culture: Spheroids were cultured in Matrigel (abs9495) for 21 days, using intestinal growth medium containing R-Spondin 1 (abs90189), Noggin (abs90191) and other factors to form induced human UC organoids (iHUCOs).

 

(3) Model characteristics

Contains epithelial and mesenchymal cell compartments that mimic the complex microenvironment of UC. Patient-specific genetic background and pathological features were preserved.

 

2.3 Evaluation indicators

 

The successful evaluation of iHUCOs is based on their ability to reproduce pathological features (histology, molecular markers), functional simulation (barrier defects, inflammatory pathways) and therapeutic responsiveness (drug intervention effect).

 

Histology and morphology: reappearance of epithelial structural abnormalities (such as stratification) in patients with UC; Goblet cells decreased and Ki67 proliferation markers significantly increased.

 

Abnormal barrier function: adhesion junctions: E-cadherin/β-catenin localized to cell membrane (healthy human-derived control organoid iHNOs distributed in cytoplasm and nucleus); Tight junction: CLDN1 expression decreased, fluorescent dextran confirmed increased permeability.

 

Molecular mechanisms: inflammatory activation: overexpression of CXCL8/CXCR1 axis and RhoA signaling; Transcriptome: Enriched for inflammation (TNF/IFN-γ), fibrosis (COL1A1/3A1) and immune infiltration (B/NK cells) pathways.

 

In vivo validation: UC phenotype preserved after transplantation; Repertaxin (abs811153, CXCR1 inhibitor) improves barrier function and reduces inflammation.

 

 

References:

[1]Chen L, Zhong XL, Cao WY, et al. IGF2/IGF2R/Sting signaling as a therapeutic target in DSS-induced ulcerative colitis. Eur J Pharmacol. 2023;960:176122.

[2]Liu W, Zhong X, Yi Y, et al. Prophylactic Effects of Betaine on Depression and Anxiety Behaviors in Mice with Dextran Sulfate Sodium-Induced Colitis. J Agric Food Chem. 2024;72(38):21041-21051. 

[3]Sarvestani SK, Signs S, Hu B, et al. Induced organoids derived from patients with ulcerative colitis recapitulate colitic reactivity. Nat Commun. 2021;12(1):262. Published 2021 Jan 11.

 

Chemical inducer of colitis model

Item number	Product Name	Specifications
abs9192	Dextran sulfate sodium salt (molecular weight: 36,000-50,000)	5g

 

Colitis positive control drug

Type of drug	Item number	Product Name	Specifications
Amino water Hydrochloric acid	abs816823	5-Aminosalicylic acid	5g
	abs813393	Sulfasalazine	1g
	abs817273	Olsalazine disodium	100mg
Adrenaline corticoids	abs813595	Dexamethasone acetate	500mg
	abs814512	Hydrocortisone acetate	100mg
	abs816630	Prednisolone acetate	500mg
Immunosuppressant	abs817118	Azathioprine	500mg
	abs814966	6-Thioguanine	500mg
	abs810016	Cyclosporine A (Cyclosporin A)	50mg

 

Traditional Chinese Medicine (Plant Extract) with Colitis Treatment Potential

Item number	Product Name	Specifications
abs47000525	Wogonin	20 mg
abs47001024	Betaine	20 mg
abs47001189	Proanthocyanidins	20 mg
abs47000689	Cyanidine chloride	20 mg
abs47001192	Proanthocyanidins B1	20 mg
abs47001193	Proanthocyanidins B2	20 mg
abs47000705	Triptolide	20 mg
abs47000092	(−)-epigallocatechin gallate	20 mg
abs47000269	Isoliquiritigenin	20 mg
abs47001134	Mullein isoflavone	20 mg
abs47000982	Silybinin	20 mg
abs47000523	Baicalin	20 mg
abs47000630	Hyperoside	20 mg
abs47000543	Astragalus polysaccharide	20 mg
abs47000469	Curcumin	20 mg
abs47000789	Rosmarinic acid	20 mg
abs47000102	Berberine	20 mg
abs47000467	Dihydromyricetin	20 mg
abs47000353	Rhein	20 mg
abs47000667	Oxymatrine	20 mg
abs47000393	Tanshinone IIA	20 mg

 

Colitis-associated agonist inhibitor

Item number	Product Name	Targets
abs810444	IWR-1 endo	Wnt/β-catenin
abs810509	IWP-2	Wnt/β-catenin
abs810420	ICG-001	Wnt/β-catenin
abs812851	Wnt Agonist 1	Wnt/β-catenin
abs810648	XAV939	Wnt/β-catenin
abs814337	NCB-0846	Wnt/β-catenin
abs817881	2-Methoxyestradiol	CYP24A1
abs815903	Ketoconazole	CYP24A1
abs816709	Clotrimazole	CYP24A1
abs810004	Genistein	CYP24A1
abs816018	vitamin D2	CYP24A1
abs812129	BI2536	PLK1
abs813185	Volasertib	PLK1
abs818402	Rigosertib	PLK1
abs813731	MLN0905	PLK1
abs817534	HMN-214	PLK1

 

Intestinal organoid culture reagent

Item number	Product name	Specifications
abs9403	Human pluripotent stem cell culture medium	500ml
abs9409	Human pluripotent stem cell digestive juice	100mL
abs9295	L-alanyl-L-glutamine solution (100 ×, 200 mM)	100mL
abs9412	ES/iPS cell cryopreservation solution	100mL
abs9410	Ready-to-use Matrigel	100mL
abs9484	RPMI 1640	500mL
abs9244	Double antibody	100mL
abs05801	Activin A	10ug
abs06269	FGF4	50ug
abs05632	WNT3a	50ug
abs9495	Matrigel (low factor, no phenol red)	1.5 ml * 8
abs945	Dialyzed fetal bovine serum	500mL
abs9545	Human normal intestinal organoid culture kit	1kit
abs812864	Y-27632	5mg
abs817694	A 83-01	5mg
abs810052	CHIR-99021	5mg
abs810009	SB202190 (FHPI)	5mg
abs816969	Nicotinamide	50mg

 

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