Composition comprising stem cell-derived exosome containing culture as effective ingredient for preventing or treating arthritis
A technology for active ingredients and arthritis, which can be used in medical preparations containing active ingredients, drug combinations, medical raw materials derived from mammals, etc., and can solve the problems of low concentration and low treatment effect of arthritis.
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[0050] Hereinafter, the present invention will be further described in detail through examples. These examples are only to illustrate the present invention more specifically, and the scope of the present invention is not limited by these examples.
[0051] Preparation Example: Stem Cells
[0052] In the following examples, the following cells were used for stem cells in order to obtain a stem cell culture medium rich in active ingredient exosomes for confirming the therapeutic effect of arthritis.
[0053] First of all, immortalized amniotic fluid stem cells (AFSC), immortalized neural stem cells (NSC) and immortalized oligodendrocyte precursor cells (OPC) are cell lines established by the inventors of the present invention as immortalized cell lines, and each cell The strains have obtained the deposit number from the Microorganism Depository Agency, and the immortalized neural stem cell CBNU-NSC cell line (preservation number: KCTC12635BP, Korea Authorized Patent No. 10-171927...
Embodiment 1
[0055] 1-1 Stem cell culture and acquisition of exosome-rich culture medium
[0056] The prepared stem cells (1x 10 6 Cells / mL) were inoculated in Dulbecco's Modified Eagle's Medium (DMEM) in 5% carbon dioxide (CO 2 ) in the supply medium for 24 hours at 37°C. Afterwards, in order to obtain exosome-enriched medium (ERCM) from stem cells, TNF-α was added to the above medium at a concentration of 50 ng / mL, and the oxygen concentration was reduced to 5%. After culturing for 24 hours, the culture medium was collected and filtered. filter, and then concentrated 10 times at low temperature to obtain the culture solution rich in exosomes derived from stem cells according to the present invention.
[0057] As a comparative example, in the process of obtaining exosome-rich culture fluid from stem cells respectively, in the process of Example 1, except for TNF-α (50ng / mL) treatment and oxygen (O 2 ) concentration was maintained at 21%, the same method as in Example 1 was carried ou...
experiment example 1
[0066] Analysis of the therapeutic effect of the present invention's stem cell-rich exosome culture fluid on arthritis
[0067] 1-1 Breeding of Rabbits for Experimental Animals
[0068] Eight-month-old (3.5kg) male New Zealand white rabbits (New Zealand white, NZW) were used after 2 weeks of laboratory acclimatization. One experimental animal was accommodated in each rabbit cage, and the environment of the animal laboratory was adjusted to a temperature of 23±2°C, a relative humidity of 55±10%, a ventilation rate of 12 times / hour, and a lighting cycle of 12 hours (07:00- 19:00), the illuminance was 150-300lux for feeding. The feed is freely ingested as a small particle type (pallet) solid feed Purina Rabbit for experimental animals and sterilized purified water. This experiment was approved by the Institutional Animal Care and Use Committee (IACUC) of the Experimental Animal Research Support Center of Chungbuk University, in accordance with the standard operation guidel...
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