Methods and systems for harvesting cells
Inventors
Kasuto, Harel • Drori-Carmi, Nirit • Zohar, Barak
Assignees
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Publication Number
US-10851341-B2
Publication Date
2020-12-01
Expiration Date
Abstract
Methods for using vibration to harvest cells grown in 3D culture are provided. The methods entail the application of force cells attached to a 3D matrix of sufficient amplitude, frequency, and duration to detach cells from the matrix and to flush the detached cells out of the matrix material. An apparatus for performing the methods of the invention as provided.
Core Innovation
The invention provides a method of seeding cells in a three-dimensional matrix in a chamber of a bioreactor. A three-dimensional matrix is provided in a fluid within a container disposed within the bioreactor chamber, and cells are introduced into the container so that the cells reside within the matrix and fluid environment.
The container is vibrated in a reciprocating motion relative to the chamber for a period of time. The vibration is performed at a frequency and amplitude sufficient to mix the cells throughout the matrix, and after mixing the vibration is discontinued to permit the cells to attach to the matrix.
The document further describes vibration-based harvesting in adherent 2D/3D matrices and associated bioreactor configurations, including open and closed bioreactor systems, wherein controlled vibratory motion is used to dissociate cells from matrices and recover released cells.
The disclosed matrices and scaffolds are suitable for adherence and vibratory interaction, including adherent fibrous matrices and biological matrices. Cell types addressed include human adherent stromal cells from placenta, adipose, and bone marrow, including fetal and maternal placenta-derived sources.
Claims Coverage
The partial content includes one independent claim. The independent claim recites four main inventive elements: providing a 3D matrix in a container within a bioreactor chamber, introducing a cell composition, vibrating the container in a reciprocating motion with frequency and amplitude sufficient to mix cells throughout the matrix, and discontinuing vibration to permit attachment.
Seeding cells into a 3-dimensional matrix in a bioreactor chamber by vibration
Providing a three-dimensional matrix in a fluid within a container disposed within a chamber of a bioreactor; introducing a composition comprising cells into the container; vibrating the container in a reciprocating motion relative to the chamber for a period of time at a frequency and amplitude sufficient to mix the cells throughout the matrix; and discontinuing the vibration to permit the cells to attach to the matrix.
Seeding adherent human cells for attachment to a 3-dimensional matrix
The method of seeding includes performing the method using adherent human cells.
Seeding adherent stromal cells from placenta, adipose, or bone marrow
The method includes adherent stromal cells originating from placenta, adipose tissue, or bone marrow.
Matrix constituents selected from enumerated adherent materials
The method includes performing seeding where the adherent material includes one or more specified materials selected from the enumerated set of polymers, biological matrices, fibers, particles, and sponges.
Reciprocating vibration with quantified amplitude and frequency constraints
The method includes a reciprocating motion having an amplitude between about 10 mm and about 750 mm and a frequency between 1 and 3 Hz.
Amplitude constrained as a fraction of container height
The method is performed such that the amplitude of a substantially linear reciprocating motion is 15-100% of the height of the container.
Overall, the claim coverage centers on vibrating a container that holds a three-dimensional matrix and cells within a bioreactor chamber to mix cells throughout the matrix, then discontinuing vibration to permit attachment. The dependent refinements further narrow cell type, matrix constituents, and vibration parameters.
Stated Advantages
Documented Applications
No documented applications found
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