US 11,920,153 B2
Compositions, devices and methods for the control in vitro of chemical microambient in cell cultures
Stefania Rapino, Bologna (IT); Francesco Zerbetto, Bologna (IT); Gastone Castellani, Bologna (IT); Stefano Salvioli, Bologna (IT); Beatrice Fraboni, Bologna (IT); Isabella Zironi, Caselecchio di Reno (IT); Maria Conte, Marzabotto (IT); Claudio Franceschi, Bologna (IT); Daniela Salvatore, Lanciano (IT); and Maila Becconi, Nuoro (IT)
Assigned to Alma Mater Studiorum—Università di Bologna, Bologna (IT)
Appl. No. 16/339,164
Filed by Alma Mater Studiorum—Università di Bologna, Bologna (IT)
PCT Filed Oct. 3, 2017, PCT No. PCT/IB2017/056081
§ 371(c)(1), (2) Date Apr. 3, 2019,
PCT Pub. No. WO2018/065887, PCT Pub. Date Apr. 12, 2018.
Claims priority of application No. 102016000099380 (IT), filed on Oct. 4, 2016.
Prior Publication US 2019/0300846 A1, Oct. 3, 2019
Int. Cl. C12N 5/00 (2006.01); C12N 5/09 (2010.01); C12N 9/00 (2006.01); C12N 9/02 (2006.01); C12N 9/04 (2006.01); C12N 9/08 (2006.01); C12N 11/04 (2006.01); C12N 11/06 (2006.01); C12N 11/087 (2020.01); C12N 11/089 (2020.01); C12N 11/12 (2006.01); G01N 33/50 (2006.01)
CPC C12N 5/0018 (2013.01) [C12N 5/0068 (2013.01); C12N 5/0693 (2013.01); C12N 9/00 (2013.01); C12N 9/0006 (2013.01); C12N 9/0036 (2013.01); C12N 9/0053 (2013.01); C12N 9/0061 (2013.01); C12N 9/0065 (2013.01); C12N 9/0069 (2013.01); C12N 9/0093 (2013.01); C12N 11/04 (2013.01); C12N 11/06 (2013.01); C12N 11/087 (2020.01); C12N 11/089 (2020.01); C12N 11/12 (2013.01); C12Y 101/03004 (2013.01); C12Y 106/03001 (2013.01); C12Y 109/03001 (2013.01); C12Y 110/03002 (2013.01); C12Y 111/01006 (2013.01); C12Y 113/12004 (2013.01); C12Y 117/03002 (2013.01); G01N 33/5008 (2013.01); C12N 2500/02 (2013.01); C12N 2501/71 (2013.01); C12N 2533/30 (2013.01); C12N 2533/50 (2013.01); C12N 2533/54 (2013.01); C12N 2533/70 (2013.01); C12N 2533/72 (2013.01); C12N 2533/74 (2013.01); C12N 2533/76 (2013.01); C12N 2533/78 (2013.01); C12N 2533/80 (2013.01); C12N 2537/10 (2013.01)] 15 Claims
OG exemplary drawing
 
1. An in vitro method to control the chemical microenvironment of a cell culture and/or mimic the physiological or pathological conditions of the cells in vivo, comprising
depositing, in a container for a cell culture, a
grid or strip(s) of a composition in the form of a hydrogel comprising a matrix that contains one or more enzymes capable of catalyzing the reduction of oxygen, so that an oxygen gradient is generated under cell cultivation from the region of the container where said hydrogel is deposited to the region of the container where said hydrogel is not deposited;
wherein said enzymes are covalently bound to said matrix by a cross-linking agent (cross-linker) or are trapped in a polymeric matrix through mechanical and/or electrostatic interactions and wherein said container is a culture plate;
cultivating a cell line or primary cells in said container.