Methods and apparatus for separating cells from microcarriers

Patent metadata

Field	Value
Publication number	US-2018142201-A1
Application number	US-201615575160-A
Country	US
Kind code	A1
Filing date	May 27, 2016
Priority date	May 29, 2015
Publication date	May 24, 2018
Grant date	—

Abstract

An apparatus for separating cells from microcarriers comprises a chamber having a body and a base. The chamber is configured to receive a mixture comprising cells and microcarriers. The apparatus further comprises a filtration element disposed within the chamber and coupled to the body. The filtration element is configured to filter microcarriers from the mixture. The apparatus further comprises an adjustment mechanism coupled to the chamber. The adjustment mechanism is configured for selective extension and retraction of at least a portion of the body of the chamber to move the filtration element relative to the base of the chamber.

First claim

The claims defining the invention are as follows:

1 . An apparatus for separating cells from microcarriers, the apparatus comprising:
a chamber having a body and a base, the chamber configured to receive a mixture comprising cells and microcarriers; a filtration element disposed within the chamber and coupled to the body, the filtration element configured to filter microcarriers from the mixture; and an adjustment mechanism coupled to the chamber, the adjustment mechanism configured for selective extension and retraction of at least a portion of the body of the chamber to move the filtration element relative to the base of the chamber.

2 . The apparatus of claim 1 , wherein selective extension of the at least a portion of the body of the chamber in response to selective actuation of the adjustment mechanism causes the at least a portion of the body to extend away from the base of the chamber and selective retraction of the at least a portion of the body of the chamber in response to selective actuation of the adjustment mechanism causes the at least a portion of the body to retract toward the base of the chamber.

3 . The apparatus of claim 1 or claim 2 , wherein selective extension of the at least a portion of the body of the chamber in response to selective actuation of the adjustment mechanism causes the filtration element to move away from the base of the chamber and selective retraction of the at least a portion of the body of the chamber in response to selective actuation of the adjustment mechanism causes the filtration element to move towards the base of the chamber.

4 . The apparatus of any one of the preceding claims, wherein the chamber is configured to transition between an extended state and a retracted state in response to selective actuation of the adjustment mechanism.

5 . The apparatus of any one of the preceding claims, wherein the filtration element comprises a filter configured to allow cells to pass through the filter and to block microcarriers from passing through the filter.

6 . The apparatus of any one of the preceding claims, wherein the filtration element comprises a filter base configured to cooperate with the base of the chamber to allow the filter base to sit flush with the base of the chamber when the apparatus is in a retracted state.

7 . The apparatus of any one of the preceding claims, wherein the body is an elongate body and the base is a conical base.

8 . The apparatus of any one of the preceding claims, wherein the body includes bellows to allow for expansion and contraction of the body.

9 . The apparatus of any one of the preceding claims, wherein the body is composed of a flexible material.

10 . The apparatus of any one of the preceding claims, wherein the adjustment mechanism comprises one or more actuators.

11 . The apparatus of claim 9 , wherein the one or more actuators comprise one or more electric cylinders.

12 . The apparatus of claim 10 or claim 11 , wherein the one or more actuators are coupled to the body of the chamber.

13 . The apparatus of any one of the preceding claims, wherein the chamber comprises a cover coupled to an first end of the body and the one or more actuators are coupled to the cover.

14 . The apparatus of any one of the preceding claims, further comprising an agitation mechanism to instigate agitation of a mixture within the chamber.

15 . The apparatus of claim 14 , wherein the agitation mechanism comprises at least one of a vibration table, a drop cam and air.

16 . A bioreactor comprising the apparatus of any one of claims 1 to 15 and an agitation system to agitate a mixture within the chamber.

17 . The bioreactor of claim 16 , wherein the agitation system comprises one or more impellers.

18 . A method for separating cells from microcarriers, the method comprising:
introducing a mixture comprising cells and microcarriers to the filtration element of the apparatus of any one of claims 1 to 15 when the apparatus is in an extended state; retaining microcarriers and cells attached to microcarriers in the filtration element; actuating the adjustment mechanism to move the filtration element toward the base of the chamber; introducing an agent for facilitating detachment of the cells from the microcarriers to the chamber; agitating the mixture in the chamber to detach at least some of the cells from the microcarriers; and retaining the microcarriers in the filtration element and allowing the detached cells and agent to pass through the filtration element.

19 . The method of claim 18 , wherein the mixture further comprises media and wherein retaining microcarriers and cells attached to microcarriers in the filtration element further comprises conveying media from the chamber.

20 . The method of claim 18 wherein the agent for facilitating detachment of the cells from the microcarriers to the chamber is introduced to the chamber via a port disposed in the base of the chamber.

21 . The method of any one of claims 18 to 20 further comprising conveying air to the chamber to assist in facilitating the mixing of the agent, cells and microcarriers.

22 . The method of any one of claims 18 to 20 further comprising conveying the agent and the detached cells from the chamber to a second chamber of a second apparatus.

23 . The method of any one of claims 18 to 22 further comprising conveying convey air to the chamber to assist in facilitating the conveying of the agent and detached cells from the chamber.

24 . The method of any one of claims 18 to 23 further comprising conveying buffer to the chamber to assist in facilitating the conveying of the agent and detached cells from the chamber.

25 . The method of any one of claims 22 to 24 further comprising introducing an agent inhibitor to the second chamber to inhibit activity of the agent.

26 . The method of any one of claims 18 to 20 further comprising:
actuating the adjustment mechanism to move the filtration element away from the base of the chamber; and
introducing an inhibitor to the chamber to inhibit activity of the agent.

27 . The method of claim 26 further comprising conveying buffer to the chamber to rinse the filtration element and chamber.

28 . The method of claim 26 or claim 27 further comprising conveying the detached cells to a collection receptacle.

29 . A method for separating cells from microcarriers, the method comprising:
introducing a mixture comprising cells and microcarriers to the filtration element of the apparatus of any one of claims 1 to 15 ; retaining microcarriers and cells attached to microcarriers in the filtration element; introducing an agent for facilitating detachment of the cells from the microcarriers to the chamber; agitating the mixture in the chamber to detach at least some of the cells from the microcarriers; retaining the microcarriers in the filtration element and allowing the detached cells and agent to pass through the filtration element; actuating the adjustment mechanism to move the filtration element away from the base of the chamber; and introducing an inhibitor to the chamber to inhibit activity of the agent.

30 . A method for separating cells from microcarriers, the method comprising:
introducing a mixture comprising cells and microcarriers to the filtration element of

Assignees

• Mesoblast Int Sarl

Inventors

• Davidzon Normand
• Simmons Paul

Classifications

• B01D35/0276

  Filtering elements with a vertical rotation or symmetry axis mounted on tanks or reservoirs · CPC title

• B01D33/01

  with translationally moving filtering elements, e.g. pistons (B01D33/04 - B01D33/327 take precedence) · CPC title

• C12M25/16

  Particles; Beads; Granular material; Encapsulation (chemical or physical processes conducted in the presence of fluids and solid particles B01J8/00) · CPC title

• C12M47/02 Primary

  Separating microorganisms from the culture medium; Concentration of biomass (separating microorganisms from their culture media C12N1/02) · CPC title

• C12M27/02

  Stirrer or mobile mixing elements · CPC title