Acellular biologic composition and method of manufacture
View Patent ↗A biological composition has a mixture of mechanically selected allogeneic biologic material derived from bone marrow. The mixture has non-whole cellular components including vesicular components and active and inactive components of biological activity, cell fragments, cellular excretions, cellular derivatives, and extracellular components. The mixture including non-whole cell fractions including one or more of exosomes, transcriptosomes, proteasomes, membrane rafts, lipid rafts. The mixture is compatible with biologic function.
1. A method of making a biological composition comprises the steps of:
collecting, recovering and processing bone marrow from cadaverous bone from a cadaver donor;
mechanically separating by spinning or tumbling cellular and non-cellular components of bone marrow from the cadaverous bone using a plurality of cycles, wherein the first cycle has a decanted fluid of unwanted cells and debris which is discarded, thereafter subsequent cycles are performed, each subsequent cycle creating a decanted fluid which is retained containing the cellular and non-cellular components, the retained decanted fluid is combined in a collection jar prior to centrifugation and passed through a blood filter to remove any bone or spicules or clumps from the cells and the non-whole cell components, wherein the non-whole cells include non-whole cellular components including vesicular components and active and inactive components of biological activity, cell fragments, cellular excretions, cellular derivatives, and extracellular components, wherein the bone marrow and cadaverous bone is discarded;
separation by density gradient centrifugation wherein the whole cells are in an interface and below;
collecting non-cellular fractions or non-cellular components or combinations thereof of predetermined density wherein fluid above the interface is removed and retained which includes non-whole cell components and excludes the whole cells;
concentrating the non-cellular fractions or non-cellular components or combinations thereof by centrifugation and filtration;
washing the non-cellular fractions or non-cellular components or combinations thereof to create a mixture;
quantifying non-whole cell fraction concentration wherein the concentration is greater than zero;
suspending the mixture to a predetermined concentration in a polyampholyte cryoprotectant;
freezing the mixture at a predetermined controlled rate.