Process and device for isolating cells from biological tissue

Patent metadata

Field	Value
Publication number	US-2017145369-A1
Application number	US-201615333337-A
Country	US
Kind code	A1
Filing date	Oct 25, 2016
Priority date	Nov 19, 2015
Publication date	May 25, 2017
Grant date	—

Abstract

The invention is directed to a Perfusion device for biological tissues comprising
a casing having two parts, a first part ( 1 ) and a second part ( 9 ), a holder ( 7 ) for a plurality of hollow penetration structures ( 8 ), wherein the hollow penetration structures ( 8 ) are provided with at least one orifice having fluid communication through the holder ( 7 ) a support ( 5 ) for the biological tissue ( 6 ) characterized in that the support ( 5 ) for the biological tissue ( 6 ) is positioned in the casing at a distance to the holder ( 7 ) that by joining the first part ( 1 ) and the second part ( 9 ) to form the casing, the hollow penetration structures ( 8 ) are in proximity to the holder ( 7 ).

Use of the perfusion device in a process for disaggregation of a biological tissue to yield target cells.

First claim

What is claimed is:

1 . A perfusion device for biological tissues, comprising:
a casing having two parts, a first part ( 1 ) and a second part ( 9 ); a holder ( 7 ) for a plurality of hollow penetration structures ( 8 ), wherein the hollow penetration structures ( 8 ) are provided with at least one orifice having fluid communication through the holder ( 7 ); and a support ( 5 ) for the biological tissue ( 6 ) characterized in that the support ( 5 ) for the biological tissue ( 6 ) is positioned in the casing at a distance to the holder ( 7 ) that by joining the first part ( 1 ) and the second part ( 9 ) to form the casing, the hollow penetration structures ( 8 ) are in proximity to the holder ( 7 ).

2 . The perfusion device according to claim 1 , characterized in that by joining the first part ( 1 ) and the second part ( 9 ) to form the casing, the hollow penetration structures ( 8 ) penetrate at least in part into the biological tissue ( 6 ).

3 . The perfusion device according to claim 1 , characterized in that the first part ( 1 ) and the second part ( 9 ) of the casing are each provided with a corresponding closing mechanism ( 11 , 12 ) for joining the two parts.

4 . The perfusion device according to claim 1 , characterized in that the hollow penetration structures ( 8 ) extend 1 to 100 mm from the holder ( 7 ).

5 . The perfusion device according to claim 1 , characterized in that the hollow penetration structures ( 8 ) are elongated objects having a needle-, cone- or pyramid-like shape.

6 . The perfusion device according to claim 1 , characterized in that the support ( 5 ) is a filter, mesh, rack, grid or a plate with orifices.

7 . The perfusion device according to claim 1 , characterized in that the holder ( 7 ) can be inserted and removed from the casing.

8 . The perfusion device according to claim 1 , characterized in that the first part ( 1 ) and the second part ( 9 ) of the casing are provided with a storage cavity ( 14 ) for fluids.

9 . The perfusion device according to claim 1 , characterized in that the first part ( 1 ) and/or the second part ( 9 ) of the casing are provided with at least one orifice for in- and/or output of fluids ( 10 , 13 ).

10 . A process for disaggregation of a biological tissue with a device according to claim 1 , characterized in that the biological tissue is penetrated at least in part by at least one of the hollow penetration structures ( 8 ) by joining the first part ( 1 ) and the second part ( 9 ) to form the casing and at least one agent to disaggregate the biological tissue into target cells is administered through the hollow penetration structures ( 8 ) into the biological tissue ( 6 ).

11 . The process according to claim 10 , characterized in that the biological tissue ( 6 ) is first washed with buffer to remove non-target cells from the biological tissue ( 6 ).

12 . The process according to claim 10 , characterized in that the agent to disaggregate the biological tissue ( 6 ) and/or the buffer is repeatedly administered into the biological tissue ( 6 ).

13 . The process according to claim 10 , characterized in that the agent to disaggregate the biological tissue ( 6 ) is selected from the group consisting of trypsin, chymotrypsin, papain, collagenase, elastase, dispase, thermolysin, hyaluronidase, clostripain and neutral protease from clostridium histolyticum , pronase, DNase I, pepsin, proteinase K, lysozyme, chelating agents for bivalent ions and mixtures thereof.

14 . The process according to claim 10 , characterized in that the target cells are tissue-resident cells.

15 . The process according to claim 10 , characterized in that the biological tissue is selected from the group consisting of spleen, heart, liver, brain and other neural tissues, kidney, lung, pancreas, breast, umbilical cord, skin, placenta, ovary, oviduct, uterus, prostate, tonsil, thymus, stomach, testis, trachea, cartilage, tendon, bone, skeletal muscle, smooth muscle, gut, colon, intestine, bladder, urethra, eye, gall bladder, organoids from cell cultures and tumors.

Assignees

• Miltenyi Biotec Gmbh

Inventors

• Poggel Carsten
• Bosio Andreas
• Stoters Wolfgang
• Adams Timo

Classifications

• C12M21/18

  Apparatus specially designed for the use of free, immobilized or carrier-bound enzymes · CPC title

• C12N5/0081

  Purging biological preparations of unwanted cells · CPC title

• C12M29/04

  Filters; Permeable or porous membranes or plates, e.g. dialysis · CPC title

• C12M29/16 Primary

  Hollow fibers (hollow fiber modules in general B01D63/02) · CPC title

• C12M23/46

  Means for fastening · CPC title