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 relates to tissue repair or regeneration using an isolated cell composition containing a mixed population of cells of hematopoietic, mesenchymal and endothelial lineage, referred to as Tissue Repair Cells (TRCs). The isolated composition is derived from mononuclear cells and provided as an exchange/washing-derived preparation described as a wash-harvest process. The process is performed in an aseptically closed chamber to reduce culture/harvest residues and mechanical damage.

The wash-harvest preparation yields a cell composition characterized by high viability and defined composition and purity parameters. The described composition shows increased post-wash viability and yield versus CYTOMATE® filtration/centrifugation, reduced residual bovine serum albumin, enrichment of CD90+ stromal and CD14auto+ monocyte/macrophage fractions, and increased VEGFR1+ cells.

Functional properties are reported as preserved clonogenic CFU-F/CFU-GM function and maintained viability after needle passage. The disclosure also describes an enhanced anti-inflammatory/immune-regulatory cytokine profile, including IL-1ra, IL-6, IL-10, MCP-1 and VEGF, with absence or low IL-12 and other TH1 cytokines, and includes inducible IDO and upregulated PD-L1 in tissue repair contexts.

Claims Coverage

The independent claim covers administration of an isolated mixed hematopoietic/mesenchymal/endothelial cell composition for tissue repair or regeneration, with 10 inventive features identified across the claim set. Dependent refinements add viability, marker composition, residue limits, contamination limits, cytokine production constraints, immunological markers, and repair targeting.

Overall claim coverage centers on administering an isolated TRC-like mixed hematopoietic/mesenchymal/endothelial cell composition defined by high viability, a specified CD90+/CD45+ distribution, low bovine serum albumin and low enzymatically active harvest reagent, and substantial freedom from mycoplasm, endotoxin and microbial contamination. Dependent refinements add quantitative pro-inflammatory cytokine production limits, IDO or PD-L1 expression, and targeting of tissue repair mechanisms such as angiogenesis and bone repair.

Stated Advantages

Documented Applications