Ex-vivo intestinal culture model, methods of producing same and uses thereof

Patent metadata

Field	Value
Publication number	US-2019100731-A1
Application number	US-201716085623-A
Country	US
Kind code	A1
Filing date	Mar 17, 2017
Priority date	Mar 17, 2016
Publication date	Apr 4, 2019
Grant date	—

Abstract

A method of preparing an ex-vivo intestinal culture model is provided. The method comprising:
(a) providing an intestinal tissue sample; (b) treating the intestinal tissue sample with a protease cleaving an extracellular matrix (ECM) component so as to break cell-cell contacts and obtain an intestinal preparation which maintains the overall intestinal tissue structure including villi and/or intestinal tissue layer orientation; (c) washing the preparation; and (d) culturing the preparation.

First claim

1 . A method of preparing an ex-vivo intestinal culture model, the method comprising:
(a) providing an intestinal tissue sample; (b) treating said intestinal tissue sample with a protease cleaving an extracellular matrix (ECM) component so as to break cell-cell contacts and obtain an intestinal preparation which maintains the overall intestinal tissue structure including villi and/or intestinal tissue layer orientation; (c) washing said preparation; and (d) culturing said preparation.

2 . (canceled).

3 . The method of claim 1 , wherein said protease comprises a protease combination comprising collagenase and dispase.

4 . The method of claim 3 , wherein said collagenase is collagenase 1A.

5 . The method of claim 3 , wherein a concentration of dispase comprises 0.05 U/ml to about 0.15 U/ml units per 3 mm 3 of the tissue.

6 . The method of claim 3 , wherein a concentration of collagenase comprises 0.1 mg/ml to about 1.0 mg/ml per 3 mm 3 of the tissue.

7 . The method of claim 1 , wherein said washing is repeated at least 2-3 times.

8 . The method of claim 1 , wherein said intestinal tissue sample is pathologic.

9 . The method of claim 7 , wherein said intestinal tissue sample comprises an inflamed tissue.

10 - 15 . (canceled)

16 . The method of claim 1 , wherein said culturing said preparation comprises placing said preparation in a culture vessel with villus surface facing up.

17 . The method of claim 1 , wherein said culturing said preparation comprises placing said preparation on a tissue culture insert.

18 . The method of claim 1 , wherein said culturing is for at least 2-4 days.

19 . An ex-vivo intestinal culture model obtainable according to the method of claim 1 .

20 . A method for screening a candidate agent for an effect on a mammalian tissue, the method comprising:
(a) contacting a candidate agent with the intestinal culture model of claim 19 , and (b) determining the effect of the agent on the intestinal culture model.

21 . A method of selecting a drug for the treatment of an intestinal disease in a subject in need thereof, the method comprising:
(a) contacting the drug with the intestinal culture model of claim 19 of the subject; and (b) determining the effect of said drug on the intestinal culture model, wherein sensitivity of said intestinal culture model to said drug indicates efficacy of said drug for the treatment of said intestinal disease.

22 . A method of treating an intestinal disease in a subject in need thereof, the method comprising:
(a) selecting a drug according to the method of claim 21 ; and (b) administering to said subject a therapeutically effective amount of a drug demonstrating efficacy for the treatment of said disease in said subject, thereby treating the disease in the subject.

23 . The method of claim 20 , further comprising stimulating an inflammatory response in said the intestinal culture model prior to or concomitantly with said contacting.

24 . The method of claim 20 , wherein said drug or agent is a drug or agent combination.

25 . The method of claim 20 , wherein said drug comprises a cannabinoid.

26 - 28 . (canceled)

29 . The method of claim 20 , wherein said determining said effect is by determining a secreted molecule.

30 . (canceled)

Assignees

• The State Of Israel Ministry Of Agriculture & Rural Development Agricultural Res Organizat
• Mor Research Applic Ltd

Inventors

• Koltai Hinanit
• Kapulnik Yoram
• Fridlender Marcelo
• Naftali Timna

Classifications

• C12N5/0679 Primary

  Cells of the gastro-intestinal tract · CPC title

• G01N33/5008

  for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics · CPC title

• C12N2509/00

  Methods for the dissociation of cells, e.g. specific use of enzymes · CPC title

• C12N2500/76

  from plants · CPC title

• G01N33/50

  Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing (measuring or testing processes involving enzymes or microorganisms, compositions or test papers therefor; processes for forming such compositions, condition responsive control in microbiological or enzymological processes C12Q) · CPC title