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 provides a composition for tissue repair that includes a bio-compatible matrix comprising a synthetic biodegradable polymeric material selected from poly-lactides, poly-glycolides, poly-fumarates and poly-ethylene glycol, and a mixed population of cells of hematopoietic, mesenchymal and endothelial lineage. The composition is defined by cell viability and a controlled immunophenotype, including at least 80% viability and about 5-75% viable CD90+ cells, with the remaining cells being CD45+.

The composition further constrains residual material by containing less than 2 μg/ml of bovine serum albumin and less than 1 μg/ml of an enzymatically active harvest reagent. The cells produce at least one anti-inflammatory cytokine selected from interleukin-1 receptor antagonist, interleukin-6, TGF-β, interleukin-8, interleukin 10, and monocyte chemoattractant protein-1, or at least one angiogenic factor, and the composition is substantially free of mycoplasm, endotoxin, and microbial contamination.

The disclosure also emphasizes a tissue-repair cell preparation characterized as substantially culture-component-free and suitable for direct human administration in physiologic solutions. A closed, automated post-culture wash/harvest separation process in the culture device minimizes mechanical damage versus conventional systems, leading to higher and more consistent viability, cell recovery, and enrichment of functional subpopulations including CD90+ stromal cells, CD14auto+ macrophages, and VEGFR1+ endothelial-supportive cells.

Claims Coverage

The provided material defines a tissue-repair composition with five inventive features: a specific polymeric matrix, a mixed lineage cell population with controlled viability and CD90+/CD45+ proportions, constrained residuals, defined cytokine and/or angiogenic factor output, and substantial freedom from contamination.

Overall, the claims center on a tissue-repair composition combining a specified synthetic biodegradable polymer matrix with a mixed hematopoietic, mesenchymal and endothelial cell population defined by viability and CD90+/CD45+ proportions, while limiting bovine serum albumin and an enzymatically active harvest reagent, enforcing functional anti-inflammatory cytokine and/or angiogenic factor production, and requiring substantial freedom from contamination.

Stated Advantages

Documented Applications