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US 10,329,530 B2Granted
Cell washing system and methods for washing small volumes of cells
AssigneeFenwal, Inc.
InventorChristopher J. Wegener
Life of this patent
Estimating the term…
Abstract
A fluid circuit for cell washing is provided that comprises a spinning membrane separator and a fluid management system comprising a cassette that defines the fluid pathways, and including internally mechanical valving, pressure sensing and air sensing for controlling flow through the fluid pathways, thus minimizing the volume of the fluid circuit. Additionally, the fluid circuit comprises syringes that are acted on by syringe pumps associated with the hardware component of the system to provide pressure for moving fluid through the circuit. Preferably, the syringes are connected directly to the cassette, or formed integrally within the cassette housing, thus further minimizing the volume of the fluid circuit.
Claims17
A disposable kit for washing a suspension of cellular material containing target and non-target components comprising:
- a) a spinning membrane separator having an inlet for flowing the suspension of cellular material to be washed and a wash medium into the spinning membrane separator, a first outlet for flowing retentate comprising a target component from the spinning membrane separator, and a second outlet for flowing filtrate comprising a non-target component of the cellular suspension and wash medium from the spinning membrane separator;
- b) a first container for receiving the retentate;
- c) a second container for receiving the filtrate;
- d) a third container of the wash medium;
- e) a first syringe comprising a barrel with a discharge port and a plunger;
- f) a second syringe comprising a barrel with a discharge port and a plunger;
- and
- g) a flow control cassette comprising a housing and having a first fluid pathway with a first inlet configured to be in fluid communication with a source of the suspension of cellular material to be washed, a second inlet configured to be in fluid communication with the third container of wash medium, and an outlet in fluid communication with the inlet of the spinning membrane separator;
- a second fluid pathway with an inlet in fluid communication with the first outlet of the spinning membrane separator for flowing retentate, a first outlet in fluid communication with the first container for receiving the retentate, and a second outlet in fluid communication with the first syringe;
- a third fluid pathway with an inlet in fluid communication with the second outlet of the spinning membrane separator for flowing filtrate, a first outlet in fluid communication with the second container for receiving the filtrate, and a second outlet in fluid communication with the second syringe;
- at least one valve or clamp located internal to the cassette for selectively occluding the fluid pathways associated with each of the first, second and third fluid pathways;
- and at least one fluid interface detector located internal to the cassette associated with each of the first, second and third fluid pathways.
9 more elements
A method for washing a suspension of cells comprising:
- mounting to a reusable hardware component a disposable kit comprising:
- a) a spinning membrane separator having an inlet for flowing the suspension of cellular material to be washed and a wash medium into the spinning membrane separator, a first outlet for flowing retentate comprising a target component from the spinning membrane separator, and a second outlet for flowing filtrate comprising a non-target component of the cellular suspension and wash media from the spinning membrane separator;
- b) a first container for receiving the retentate;
- c) a second container for receiving the filtrate;
- d) a third container of the wash media;
- e) a first syringe comprising a barrel with a discharge port and a plunger;
- f) a second syringe comprising a barrel with a discharge port and a plunger;
- and
- g) a flow control cassette comprising a housing and having a first fluid pathway with a first inlet configured to be in fluid communication with a source of the suspension of cellular material to be washed, a second inlet configured to be in fluid communication with the third container of wash media, and an outlet in fluid communication with the inlet of the spinning membrane separator;
- a second fluid pathway with an inlet in fluid communication with the first outlet of the spinning membrane separator for flowing retentate, a first outlet in fluid communication with the first container for receiving the retentate, and a second outlet in fluid communication with the first syringe;
- a third fluid pathway with an inlet in fluid communication with the second outlet of the spinning membrane separator for flowing filtrate, a first outlet in fluid communication with the second container for receiving the filtrate, and a second outlet in fluid communication with the second syringe;
- at least one valve or clamp located internal to the cassette for selectively occluding the fluid pathways associated with each of the first, second and third fluid pathways;
- and at least one fluid interface detector located internal to the cassette associated with each of the first, second and third fluid pathways;
- priming the disposable kit with wash media;
- loading the spinning membrane separator with a volume of cellular material to be washed;
- washing the cellular material by introducing wash media to the spinning membrane separator to remove supernatant and non-target materials from the spinning membrane separator;
- clearing the spinning membrane separator of washed cells;
- and
- flowing the washed cells to the container for receiving retentate;
- wherein the priming, loading, washing, clearing and flowing steps are controlled by a programmable controller based on signals received from the fluid interface detectors associated with each of the first, second and third fluid pathways.
17 more elements
Dependencies are inferred from the claim text.