Aseptic tissue processing method, kit and device

Abstract

The present invention concerns a single use aseptic kit comprising: a disaggregation module for receipt and processing of material comprising solid mammalian tissue; and a stabilisation module for storing disaggregated product material, wherein each of the modules comprises one or more flexible containers connected by one or more conduits adapted to enable flow of the tissue material there between; and wherein each of the modules comprises one or more ports to permit aseptic input of media and/or reagents into the one or more flexible containers. The invention further relates to an automated device for semi-automated aseptic disaggregation and/or enrichment and/or stabilisation of cells or cell aggregates from mammalian solid tissue comprising a programmable processor and the single use aseptic kit. The invention further relates to a semi-automatic aseptic tissue processing method.

1 : A single use aseptic kit comprising:
 a disaggregation module for receipt and processing of material comprising solid mammalian tissue; and   a stabilisation module for further processing and/or storing disaggregated product material,   wherein each of said modules comprises one or more flexible containers connected by one or more conduits adapted to enable flow of the tissue material there between, and   wherein each of said modules comprises one or more ports to permit aseptic input of media and/or reagents into the one or more flexible containers,   wherein the kit further comprises a digital, electronic, or electromagnetic tag identifier relating to a specific program that defines a type of disaggregation, enrichment, and/or stabilisation process and use of one or more types of media in the process(es).   
     
     
         2 : The single use aseptic kit of  claim 1 , wherein the one or more flexible containers comprise a resilient deformable material. 
     
     
         3 : The single use aseptic kit of  claim 1 , wherein the one or more flexible containers of the disaggregation module comprises one or more sealable openings. 
     
     
         4 : The single use aseptic kit of  claim 3 , wherein the flexible container of the disaggregation module comprises a heat sealable weld. 
     
     
         5 : The single use aseptic kit of  claim 1 , wherein the one or more flexible containers comprises internally rounded edges. 
     
     
         6 : The single use aseptic kit of  claim 1 , wherein the one or more flexible containers of the disaggregation module comprises disaggregation surfaces adapted to mechanically crush and shear the solid tissue therein. 
     
     
         7 : The single use aseptic kit of  claim 1 , further comprising an enrichment module comprising one or more flexible containers, wherein the enrichment module is configured for filtration of disaggregated solid tissue material and segregation of non-disaggregated tissue and filtrate and comprising, wherein the enrichment module is connected to the stabilisation and desegregation modules by one or more conduits adapted to enable flow of the tissue material, and wherein the one or more flexible containers of the enrichment module each comprise a filter that is configured to retain a retentate of cellularised disaggregated solid tissue. 
     
     
         8 : The single use aseptic kit of  claim 1 , wherein the one or more flexible containers of the stabilisation module comprises media formulation for storage of viable cells in solution or in a cryopreserved state. 
     
     
         9 . (canceled) 
     
     
         10 . (canceled) 
     
     
         11 : The single use aseptic kit of  claim 7 , wherein the same flexible container forms part of one or more of the disaggregation modules, stabilisation modules, and/or enrichment modules. 
     
     
         12 : The single use aseptic kit of  claim 11 , wherein the disaggregation module comprises a first flexible container for receipt of the tissue to be processed. 
     
     
         13 : The single use aseptic kit of  claim 12 , wherein the disaggregation module comprises a second flexible container comprising the media for disaggregation. 
     
     
         14 : The single use aseptic kit of  claim 13 , wherein the enrichment module comprises a first flexible container and a third flexible container configured to receive enriched filtrate. 
     
     
         15 : The single use aseptic kit of  claim 14 , wherein both the disaggregation module and the stabilisation module comprise a second flexible container and wherein the second container comprises digestion media and stabilisation media. 
     
     
         16 : The single use aseptic kit of  claim 15 , wherein the stabilisation module comprises a fourth flexible container comprising stabilisation media. 
     
     
         17 : The single use aseptic kit of  claim 16 , wherein the stabilisation module also comprises the first flexible container and/or the third flexible container for storing and/or undergoing cryopreservation. 
     
     
         18 : Use of the single use aseptic kit according to  claim 1  in a semi-automated process for the aseptic disaggregation, stabilisation, and enrichment of mammalian cells or cell aggregates. 
     
     
         19 : An automated device for semi-automated aseptic disaggregation, enrichment, and/or stabilisation of cells or cell aggregates from mammalian solid tissue comprising:
 a programmable processor; and   a radio-frequency identification tag reader,   wherein the programmable processor is configured to recognise a single-use aseptic kit in accordance with  claim 1  using the tag reader and is adapted to subsequently execute a kit-specific program, wherein the program defines the type of disaggregation, enrichment, and/or stabilisation processes and the media type(s) required for use in the process(es).   
     
     
         20 - 21 . (canceled) 
     
     
         22 : The automated device of  claim 19 , wherein the programmable processor is adapted to communicate with and control one or more of:
 the disaggregation module;   the enrichment module; and   the stabilisation module.   
     
     
         23 : The automated device of  claim 22 , wherein the programmable processor controls the disaggregation module to enable a physical and/or biological breakdown of the solid tissue material. 
     
     
         24 : The automated device of  claim 23 , wherein the programmable processor controls the disaggregation module to enable a physical and enzymatic breakdown of the solid tissue material. 
     
     
         25 : The automated device of  claim 24 , wherein the enzymatic breakdown of the solid tissue material is by one or more media enzyme solutions selected from collagenase, trypsin, lipase, hyaluronidase, deoxyribonuclease, Liberase H1, pepsin, or mixtures thereof. 
     
     
         26 : The automated device of  claim 19 , wherein the programmable processor controls disaggregation surfaces within the disaggregation flexible containers that mechanically crush and shear the solid tissue and wherein the disaggregation surfaces are mechanical pistons. 
     
     
         27 : The automated device of  claim 19 , wherein the programmable processor controls the stabilisation module to cryopreserve the enriched disaggregated solid tissue in the container using a programmable temperature. 
     
     
         28 : The automated device of  claim 19  wherein the device further comprises one or more of the additional components in any combination:
 sensors capable of recognising whether a disaggregation process has been completed in the disaggregation module prior to transfer of the disaggregated solid tissue to the enrichment module; 
 weight sensors configured to determine an amount of media required in the containers of one or more of the disaggregation module; the enrichment module; and/or the stabilisation module and to control the transfer of material between respective containers; 
 sensors to sense temperature within the containers of the one or more of the disaggregation module; the enrichment module; and/or 
 the stabilisation module; 
 at least one bubble sensor to control the transfer of media between the input and output ports of each container in the module; 
 at least one peristaltic pump to control the transfer of media between the input and output ports; 
 pressure sensors to assess the pressure within the enrichment module; 
 one or more valves to control a tangential flow filtration process within the enrichment module; and/or 
 one or more clamps to control the transfer of media between the input and output ports of each module. 
 
     
     
         29 : The automated device of  claim 19 , wherein the programmable processor is adapted to maintain an optimal storage temperature range in the stabilisation module until the container is removed; or executes a controlled freezing step. 
     
     
         30 : The automated device of  claim 19 , further comprising a user interface. 
     
     
         31 : The automated device of  claim 23 , wherein the interface comprises a display screen to display instructions that guide a user to input parameters, confirm pre-programmed steps, warn of errors, or combinations thereof. 
     
     
         32 : The automated device of  claim 19 , wherein the automated device is adapted to be transportable. 
     
     
         33 : A semi-automatic aseptic tissue processing method comprising:
 automatically determining aseptic disaggregation tissue processing steps and their associated conditions from a digital, electronic, or electromagnetic tag identifier associated with the aseptic processing kit of  claim 1 ;   placing a tissue sample into a flexible plastic container of the disaggregation module of the aseptic processing kit; and   processing the tissue sample by automatically executing the one or more tissue processing steps by communicating with and controlling the disaggregation module; the optional enrichment module; and the stabilisation module.   
     
     
         34 : The single use aseptic kit of  claim 1 , further comprising an enrichment module for filtration of disaggregated solid tissue material and segregation of non-disaggregated tissue and filtrate. 
     
     
         35 : The single use aseptic kit of  claim 1 , wherein the specific program defines use of a freezing solution for controlled rate freezing in the stabilisation module.