Methods for isolating umbilical cord blood plasma products, tissue and cellular exosomes, and compositions and methods of use thereof
AssigneeRutgers, The State University of New Jersey
InventorsWise Young, Dongming Sun
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Abstract
Described herein are novel methods for fractionating and isolating platelets, platelet- and extracellular vesicle-derived growth factors, exosomes, globulins, fibrinogen and albumin, and methods of using the isolated platelets, platelet and extracellular vesicle-derived growth factors, exosomes, globulins, fibrinogen and albumin for regenerating tissue in a subject, treating fibrinogenemia or a clotting deficiency in a subject, treating ischemia and hypoxia, treating dry-eye syndrome, an orthopedic disorder, or a dental disorder in a subject. Also described herein are growth media for culturing mammalian (e.g., human) cells.
Claims18
A method of isolating one or more of:
- exosomes, globulins, fibrinogen, and albumin from a tissue source, the method comprising:
- i. providing a tissue source of exosomes, globulins, fibrinogen, and albumin;
- ii. subjecting the tissue source to filtration with a micropore filter comprising pores that are 0.22 μm in diameter resulting in a plasma filtrate comprising exosomes, globulins, fibrinogen, albumin, and citrate-phosphate-dextrose (CPD);
- iii. adding the plasma filtrate to a solution comprising ultrasonicated polyethylene glycol (PEG) bubbles that are less than 340 nm in diameter and transferring the solution to a first centrifuge tube such that the exosomes partition into PEG bubbles;
- iv. cooling the first centrifuge tube to 2° C. to 8° C. for 8 hours or more;
- V. centrifuging the first centrifuge tube such that the exosomes and the PEG form a first pellet, and a first supernatant is formed, the first supernatant comprising albumin, globulins, CPD, and fibrinogen;
- and
- vi. recovering the first supernatant and freezing the first pellet comprising exosomes and PEG for storage;
- wherein about 80% to about 95% of the exosomes from umbilical cord blood (UCB) are isolated.
Dependencies are inferred from the claim text.