Milk isolated powdered exosomes, nutritional compositions and methods
A technology for nutritional compositions and exosomes, which are used in drug combinations, endocrine system diseases, medical preparations containing active ingredients, etc.
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Embodiment 1
[0066] Example 1: Production of powdered exosomes
[0067] This example describes the preparation of powdered exosomes and powdered nutritional compositions comprising exosomes. Exosomes are first isolated from bovine milk and then dried. Exosomes were isolated from bovine milk using the following procedure. Raw raw milk was aliquoted and immediately frozen at -80°C. Aliquots were thawed on ice and subjected to a first centrifugation at about 12,000G for about 15 minutes at about 4°C to obtain a whey layer formed between the fat (lipid) top layer and the cell debris pellet. The whey fraction was transferred to a clean tube and centrifuged two more times at about 12,000G and about 4°C for about 30 minutes each to remove extra fat and debris. A substantially clarified whey fraction is obtained. The substantially clarified whey fraction was microfiltered using a 0.22 μm filter of hydrophilic polyethersulfone, and the microfiltered whey was centrifuged at 100,000 G for approxi...
Embodiment 2
[0079] Example 2: Treatment Results
[0080] This example demonstrates that intact powdered exosomes were prepared and shown to be taken up by muscle cells as described above. This was shown by labeling exosomes with a red dye, incubating the exosomes with L6.C11 muscle cells, and observing the cells by microscopy. A control comprising L6.C11 muscle cells incubated with BSA labeled with the same red dye demonstrated that the dye itself did not promote uptake.
[0081] Powdered exosomes lead to increased glucose uptake by muscle cells. Powdered exosomes were added to L6.C11 muscle cell cultures until reaching milk exosome concentrations of 0, 2.8, 5.6 and 14 μg / ml, respectively, and the cultures were incubated for 24 hours. Uptake of radiolabeled glucose was then assessed. As a result of these incubations, the image 3The white bar displays. A concentration of 0 μg / ml (no exosomes) served as a control, where glucose uptake was set at 100%. Concentrations of 5.6 and 14 μg / ...
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Abstract
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