Mixed cell populations for tissue repair and separation technique for cell processing

Inventors

Hampson, Brian • Goltry, Kristin • Smith, Douglas M. • Rowley, Jonathan A. • Venturi, Naia

Assignees

Vericel Corp

Abstract

The present invention provides a fluid exchange cell culture technique and tissue repair cells (TRCs) made by these methods, as well as methods using these cells. The method includes a new wash step which increases the tissue repair properties of the TRCs of the invention. This wash step allows for the production of TRC populations with greater tissue repair and anti-inflammatory capabilities. Embodiments of the present invention include a post-culture process for cultured cells that preferably includes the steps of: a wash process for removing unwanted residual culture components, a volume reduction process, and a harvesting process to remove cultured cells. Preferably, all these steps are performed within a aseptically closed cell culture chamber by implementing a separation method that minimizes mechanical disruption of the cells and is simple to automate. The harvested cells may then be concentrated to a final volume for the intended use. In such embodiments, the final composition is a substantially purified and concentrated cell mixture suspended in a physiologic solution suitable for immediate use in humans without further washing, volume reduction, or processing. Embodiments are also applicable to harvesting (and/or washing) particles within a liquid or solution within a chamber.

Core Innovation

The invention describes processing tissue repair cells (TRCs) in an aseptically closed biochamber using a wash-harvest workflow. The workflow displaces culture media from the biochamber into a waste container by introducing a biocompatible first rinse solution, replaces the first rinse with a cell harvest solution comprising an enzyme that allows detachment of cells from each other and/or from the biochamber surface, and incubates to allow detachment while keeping the cells settled on the surface.

The detached cells are then kept settled while the cell harvest solution is displaced into the waste container using a second rinse solution introduced into the biochamber. After the second rinse displacement, the biochamber is agitated to bring the settled cells into suspension for collection as a mixed cell population. The method results in a mixed cell population that is at least 80% viable and at least 5% of the cells express CD90, with the compositions further characterized by CD90+ enrichment and low residual culture components.

The resulting mixed population includes an anti-inflammatory/immunomodulatory cytokine profile and low/undetectable IL-12 and inducible IDO and PD-L1 as described in the provided content. The described wash-harvest approach is contrasted with CYTOMATE® washing, and the document reports improvements in post-wash and post-storage viability/yield, increased CD90+ content, increased CD14auto+ and VEGFR1+ fractions, improved CFU-F/CFU-GM per dose, and maintenance of viability after needle passage.

Claims Coverage

Independent claim clm-00001 defines a biochamber-based culture and wash-harvest method with enzyme detachment while keeping cells settled, followed by agitation to resuspend, producing a mixed cell population defined by viability and CD90 expression. Additional inventive features include specific cell sources, a human-injectable second rinse solution, gas-assisted reduced liquid volume, a flow-rate range for introducing solutions and gases, and a radial plug flow arrangement.

Overall, claim coverage centers on a wash-harvest processing method in a biochamber that uses first and second rinses with an enzyme harvest step, while maintaining cells in a settled state during detachment and rinse displacement, then agitating to resuspend to yield a mixed cell population defined by viability and CD90 expression. Dependent claims further define cell sourcing, human-injectable second rinse suitability, gas-assisted reduced liquid volume, specified flow-rate limits, and a radial plug flow introduction pattern.

Stated Advantages

Documented Applications