METHODS, SYSTEMS AND APPARATUS FOR ISOLATING EXOSOMES FROM A BIOLOGICAL SAMPLE
Described herein are methods, systems and apparatus for separating components of a biological sample; and in particular, extracting and isolating exosomes. Methods of administering the exosomes are also described.
1 . A method for:
separating and/or isolating exosomes in a post-centrifuged biological sample;
increasing the number of exosomes available for administration to a patient; and/or
enriching the exosome count in a biological sample, comprising:
centrifuging a collection tube containing a biological sample and a separation barrier (e.g., a float or a thixotropic gel); and
agitating the collection tube at an angle and rate effective to create a visually perceivable layer of foam on top of said biological sample.
2 . The method according to claim 1 , wherein the collection tube further comprises an anticoagulant.
3 . The method according to claim 1 , wherein the biological sample comprises a plurality of components.
4 . The method according to claim 1 , wherein the centrifugation is performed at a force of from about 500 g up to about 4000 g for a time sufficient to separate the plurality of components in the biological sample.
5 . The method according to claim 1 , wherein the thixotropic gel forms a barrier between the plurality of components of the biological sample.
6 . The method according to claim 1 , wherein the biological sample comprises whole blood.
7 . The method according to claim 1 , further comprising the step of activating the platelets to induce the release of exosomes.
8 . The method according to claim 7 , wherein the activating step comprises the application of a chemical activating agent.
9 . The method according to claim 7 , wherein the activating step does not involve a chemical activating agent.
10 . The method according to claim 8 , wherein the chemical activating agent is selected from:
calcium chloride, calcium gluconate, thrombin, collagen, lipopolysaccharide, Ca 2+ ionophores;
and a combination of two or more thereof.
11 . The method according to claim 9 , wherein the activating step comprises applying energy to the biological sample.
12 . The method according to claim 11 , wherein the energy comprises light energy or electromagnetic energy.
13 . The method according to claim 12 , wherein the energy comprises infrared light.
14 . The method according to claim 1 , further comprising extracting at least a portion of the biological sample comprising exosomes.
15 . The method according to claim 14 , further comprising purifying the sample to isolate the exosomes.
16 . The method according to claim 15 , wherein the purification step comprises filtering the portion of the biological sample comprising exosomes.
17 . A method for treating, preventing or ameliorating a symptom associated with alopecia, bed sores, wrinkles, pain, tendonitis, arthritis, acne, scarring, crow's feet, ligament sprains and tears, and/or skin lesions comprising administering a biological sample according to claim 14 , to a subject in need thereof.
18 . The method according to claim 17 , wherein the administering step comprises application of the biological sample comprising exosomes to a skin surface of the subject in need thereof.
19 . The method according to claim 18 , wherein the biological sample comprising exosomes is administered via injection or via mask.