Chemical testing

Patent metadata

Field	Value
Publication number	US-2018024121-A1
Application number	US-201515536558-A
Country	US
Kind code	A1
Filing date	Dec 18, 2015
Priority date	Dec 18, 2014
Publication date	Jan 25, 2018
Grant date	—

Abstract

The present invention relates a method for chemical testing, comprising culturing cells in a first plant-derived nanofibrillar cellulose (NFC) hydrogel to obtain in vivo like cells; exposing the in vivo like cells to a test chemical;
optionally within another plant-derived NFC hydrogel; incubating the exposed in vivo like cells; detecting during or after incubating, the impact of the test chemical on the in vivo like cells by at least one detection; and removing the plant-derived NFC hydrogel at least once at any stage after obtaining the in vivo like cells and before at least one detection used for detecting the impact of the test chemical on the in vivo like cells. The invention further relates to the use of plant-derived NFC hydrogel in a method for chemical testing, the use of in vivo like cells obtained by culturing cells in plant-derived NFC hydrogel for chemical testing and to a kit for chemical testing comprising plant-derived NFC hydrogel, instructions and a cell or test chemical library.

First claim

1 . A method for chemical testing, comprising
a) culturing cells on or in a first plant-derived nanofibrillar cellulose (NFC) hydrogel to obtain in vivo like cells; b) exposing the in vivo like cells to a test chemical, optionally within another plant-derived NFC hydrogel; c) incubating the exposed in vivo like cells; d) detecting during or after the incubating impact of the test chemical on the in vivo like cells by at least one detection; and e) removing the plant-derived NFC hydrogel at least once at any stage after stage a) and before at least one detection according to stage d).

2 . The method according to claim 1 further comprising removing the plant-derived NFC hydrogel between a) and b).

3 . The method according to claim 1 further comprising removing the plant-derived NFC hydrogel between b) and d).

4 . The method according to claim 1 , further comprising exposing the in vivo like cells to a test chemical within the hydrogel of stage a) or exposing the in vivo like cells to a test chemical within another NFC hydrogel having different properties than the NFC hydrogel of stage a).

5 . The method according to claim 4 , wherein said different properties relate to one or more properties chosen from stiffness of NFC hydrogel, NFC concentration, shear-zero viscosity of NFC hydrogel, charge of NFC, and transparency of NFC hydrogel.

6 . The method according to claim 1 , wherein said chemical testing comprises toxicity testing; safety testing; drug candidate testing; drug screening; or pro-drug candidate testing.

7 . The method according to claim 1 , wherein said test chemical is selected from the group consisting of drugs, drug candidates, pro-drugs, pro-drug candidates, nanoparticles, cell regulatory agents, food or food additives, household products, industrial chemicals, packing materials, air freshener, plant growth regulatory agents, environmental toxins, pesticides, personal care products, or their chemical ingredients.

8 . The method according to claim 1 , wherein said in vivo like cells are stem cells, primary cells or secondary cells or any combinations thereof.

9 . The method according to claim 1 , wherein the NFC hydrogel is provided onto a support before cells for culturing are seeded onto or into the hydrogel; or after cells for culturing are first seeded into the hydrogel.

10 . The method according to claim 1 , wherein the removing the plant-derived NFC hydrogel comprises physical, mechanical or chemical removing or any combination thereof, the plant-derived NFC being treated enzymatically with a cellulase for a time sufficient to at least partly release the cells.

11 . The method according to claim 10 , wherein the cellulase is a cellulolytic enzyme mixture, optionally comprising hemicellulases, a commercial cellulose, a partially purified cellulose, or a purified cellulase.

12 . The method according to claim 1 , wherein the detection of the impact of the test chemical on the in vivo like cells comprises qualitative detecting, quantitative detecting, or any combinations thereof.

13 . The method according to claim 1 , wherein the detection of the impact of the test chemical on the in vivo like cells comprises a chromatographic detection system, an optical detection system, or a combination thereof.

14 . The method according to claim 13 wherein the chromatographic detection system is based on chromatography such as GC, HPLC, affinity, displacement, ion-exchange, size exclusion, gel-filtration, fast protein liquid, paper, or thin-layer chromatography; or on electrochromatography such as gel-electrophoresis, 2D gel-electrophoresis, or isoelectric focusing.

15 . The method according to claim 13 wherein the optical detection system is based on one or more of visual examination; spectroscopy such as nuclear magnetic resonance (NMR), Raman, IR, UV, visible light, fluorescence, mass spectrometry (MS); microscopy such as optical microscopy (phase contrast, reverse phase contrast, confocal, fluorescence) or electron microscopy (TEM, SEM) or scanning-probe microscopy (AFM); photometry; laser or flow-cytometry, optionally using high content screening and/or isotope labelling.

16 . The method according to claim 1 , wherein the detection of the impact of the test chemical on the in vivo like cells further comprises transplantation of the exposed in vivo like cells into test animals, optionally followed by preparation and detecting of histological samples.

17 . The method according to claim 1 , wherein said NFC hydrogel includes from 0.05 to 10 wt % of nanofibrillar cellulose.

18 . The method according to claim 1 , wherein said NFC hydrogel comprises native nanofibrillar cellulose or anionic nanofibrillar cellulose.

19 . The method according to claim 1 , wherein said NFC hydrogel comprises nanofibrillar cellulose including cellulose I.

20 . The method according to claim 1 , wherein said NFC hydrogel has a stiffness of at least 4 Pa.

21 . Use of plant-derived NFC hydrogel in a method for chemical testing, wherein a NFC hydrogel is used for culturing cells on or in said hydrogel to obtain in vivo like cells; said in vivo like cells are exposed to a test chemical; the exposed in vivo like cells are incubated; and the impact of the test chemical on the in vivo like cells is detected by at least one detection during or after the incubation; wherein the plant-derived NFC hydrogel is removed at any stage after obtaining the in vivo like cells and before a last detection of the impact of the test chemical on the in vivo like cells.

22 . Use of in vivo like cells obtained by culturing cells on or in plant-derived NFC hydrogel for chemical testing, which use comprises removing the plant-derived NFC hydrogel before detecting the impact of a test chemical on the in vivo like cells.

23 . A kit for chemical testing comprising plant-derived NFC hydrogel, the kit comprising:
instructions for carrying out the method according to claim 1 ; and at least one library chosen from a cell library and a test chemical library.

24 . The kit according to claim 23 characterized in that the NFC hydrogel is provided in a form of an aqueous stock comprising from 0.5 to 25 w % of NFC.

25 . The kit according to claim 23 , characterized in that the plant-derived NFC hydrogel is on a support.

26 . The kit according to claim 23 , further comprising a vial of cellulase.

27 . The kit according to claim 23 , characterized in that the test chemical library is added to the NFC hydrogel.

28 . The kit according to claim 23 , characterized in that the cell library comprises cultured in vivo like cells, or cells seeded in the NFC hydrogel.

Assignees

• Upm Kymmene Corp

Inventors

• Yliperttula Marjo
• Urtti Arto
• Lou Yan-Ru
• Malinen Melina
• Laurén Patrick
• Kanninen Liisa
• Niklander Johanna
• Somersalo Petter
• Harjumäki Riina

Classifications

• G01N33/5082 Primary

  Supracellular entities, e.g. tissue, organisms · CPC title

• G01N33/5023

  on expression patterns · CPC title

• G01N33/5026

  on cell morphology · CPC title

• G01N33/5014

  for testing toxicity · CPC title

• G01N33/5436

  with ligand physically entrapped within the solid phase (liposomes G01N33/5432; immunological test elements G01N33/54386) · CPC title