Pharmaceutical composition and cosmetic composition

Abstract

To provide pharmaceutical compositions and cosmetic compositions that effectively utilize stem cells and stem cell culture media.SOLUTION: A fat-decomposing agent, an agent for promoting induction of differentiation of a myotube from a myoblast, a prostacyclin production promoter, a hyaluronic acid production promoter for a chondrocyte, an anti-aging agent, an agent for treating or preventing a lifestyle-related disease, or an agent for treating or preventing an adult disease each comprises stem cells, an extract of stem cells, a medium of stem cells, a supernatant of a culture medium during stem cell induction, a supernatant of a culture medium of stem cells, or a secretion of stem cells.SELECTED DRAWING: None

Description

[Technical field]

[0001] The present invention relates to pharmaceutical and cosmetic compositions. [Background technology]

[0002] Embryonic stem cells (ES cells) are stem cells established from early human or mouse embryos. ES cells have the pluripotency to differentiate into all cells present in the body. Currently, human ES cells are available for cell transplantation therapy for many diseases, including Parkinson's disease, juvenile diabetes, and leukemia. However, there are obstacles to the transplantation of ES cells. In particular, the transplantation of ES cells can provoke an immune rejection response similar to that which occurs following unsuccessful organ transplants. In addition, there is much criticism and opposition from an ethical standpoint to the use of ES cells, which are established by destroying human embryos.

[0003] Under these circumstances, Professor Shinya Yamanaka of Kyoto University succeeded in establishing induced pluripotent stem cells (iPS cells) by introducing four genes, OCT3 / 4, KLF4, c-MYC, and SOX2, into somatic cells. This led to Professor Yamanaka receiving the Nobel Prize in Physiology or Medicine in 2012 (see, for example, Patent Document 1). iPS cells are ideal pluripotent cells that are free from rejection reactions and ethical issues. Therefore, iPS cells are expected to be used in cell transplantation therapy. Meanwhile, there have been reports of reusing media used to culture iPS cells into pharmaceutical compositions (see, for example, Patent Document 2). [Prior art documents] [Patent documents]

[0004] [Patent Document 1] Patent No. 4183742 [Patent Document 2] JP 2016-128396 A Summary of the Invention [Problem to be solved by the invention]

[0005] One of the objects of the present invention is to provide pharmaceutical and cosmetic compositions that effectively utilize stem cells and stem cell culture media. [Means for solving the problem]

[0006] According to an aspect of the present invention, there is provided a lipolytic agent comprising stem cells, a stem cell extract, stem cell culture medium, a culture supernatant during stem cell induction, a culture supernatant of stem cell, or a secretion product of stem cells.

[0007] According to an aspect of the present invention, there is provided an anti-obesity drug comprising stem cells, a stem cell extract, stem cell culture medium, a culture medium supernatant during stem cell induction, a culture medium supernatant of stem cells, or a secretion product of stem cells.

[0008] According to an aspect of the present invention, there is provided an obesity-improving drug comprising stem cells, a stem cell extract, stem cell culture medium, a culture supernatant during stem cell induction, a culture supernatant of stem cell, or a secretion product of stem cells.

[0009] According to an aspect of the present invention, there is provided a fat decomposition promoter comprising stem cells, a stem cell extract, a stem cell culture medium, a culture medium supernatant during stem cell induction, a culture medium supernatant of stem cells, or a secretion product of stem cells.

[0010] According to an aspect of the present invention, there is provided a dietary agent comprising stem cells, a stem cell extract, a stem cell culture medium, a culture supernatant during stem cell induction, a culture supernatant of stem cells, or a secretion product of stem cells.

[0011] According to an aspect of the present invention, there is provided a drug for treating or preventing diabetes, comprising stem cells, a stem cell extract, a stem cell culture medium, a culture medium supernatant during stem cell induction, a stem cell culture medium supernatant, or a stem cell secretion product.

[0012] According to an aspect of the present invention, there is provided an agent for promoting differentiation induction from myoblasts to myotubes, comprising stem cells, a stem cell extract, a stem cell medium, a supernatant of a culture medium during stem cell induction, a supernatant of a culture medium of stem cells, or a secretion product of stem cells.

[0013] According to an aspect of the present invention, there is provided a muscle formation promoter comprising stem cells, a stem cell extract, a stem cell culture medium, a culture supernatant during stem cell induction, a culture supernatant of stem cell culture medium, or a stem cell secretion product.

[0014] According to an aspect of the present invention, there is provided an agent for maintaining muscle mass, comprising stem cells, a stem cell extract, a stem cell culture medium, a culture supernatant during stem cell induction, a culture supernatant of stem cells, or a secretion product of stem cells.

[0015] According to an aspect of the present invention, there is provided an agent for increasing muscle mass, comprising stem cells, a stem cell extract, a stem cell culture medium, a culture supernatant during stem cell induction, a culture supernatant of stem cells, or a secretion product of stem cells.

[0016] According to an aspect of the present invention, there is provided a prostacyclin production promoter comprising stem cells, a stem cell extract, a stem cell culture medium, a culture supernatant during stem cell induction, a culture supernatant of a stem cell culture medium, or a secretion product of a stem cell.

[0017] According to an aspect of the present invention, there is provided an adenylate cyclase activator comprising stem cells, a stem cell extract, stem cell culture medium, a culture supernatant during stem cell induction, a culture supernatant of stem cells, or a secretion product of stem cells.

[0018] According to an aspect of the present invention, there is provided a cyclic adenosine monophosphate production promoter comprising stem cells, a stem cell extract, a stem cell culture medium, a culture medium supernatant during stem cell induction, a culture medium supernatant of a stem cell, or a stem cell secretion product.

[0019] According to an aspect of the present invention, there is provided a platelet aggregation inhibitor comprising stem cells, a stem cell extract, stem cell culture medium, a culture medium supernatant during stem cell induction, a culture medium supernatant of stem cells, or a stem cell secretion product.

[0020] According to an aspect of the present invention, there is provided a vasodilator comprising stem cells, a stem cell extract, stem cell culture medium, a culture supernatant during stem cell induction, a culture supernatant of stem cell, or a secretion product of stem cells.

[0021] According to an aspect of the present invention, there is provided a drug for treating or preventing hypertension, comprising stem cells, a stem cell extract, a stem cell culture medium, a culture medium supernatant during stem cell induction, a stem cell culture medium supernatant, or a stem cell secretion product.

[0022] According to an aspect of the present invention, there is provided a therapeutic or preventive agent for arteriosclerosis, comprising stem cells, a stem cell extract, a stem cell culture medium, a culture medium supernatant during stem cell induction, a culture medium supernatant of stem cells, or a secretion product of stem cells.

[0023] According to an aspect of the present invention, there is provided an agent for inhibiting vascular endothelial cell proliferation, comprising stem cells, a stem cell extract, a stem cell culture medium, a culture medium supernatant during stem cell induction, a culture medium supernatant of stem cells, or a secretion product of stem cells.

[0024] According to an aspect of the present invention, there is provided a chondrocyte hyaluronic acid production promoter, comprising stem cells, a stem cell extract, a stem cell medium, a culture medium supernatant during stem cell induction, a culture medium supernatant of stem cells, or a secretion product of stem cells. The chondrocyte hyaluronic acid production promoter may further comprise hyaluronic acid.

[0025] According to an aspect of the present invention, there is provided a chondrocyte protective agent or proliferation promoter, comprising stem cells, a stem cell extract, a stem cell medium, a culture supernatant during stem cell induction, a culture supernatant of stem cells, or a secretion product of stem cells. The chondrocyte protective agent or proliferation promoter may further comprise hyaluronic acid.

[0026] According to an aspect of the present invention, there is provided a therapeutic or preventive agent for wounds of cartilage tissue, comprising stem cells, a stem cell extract, a stem cell medium, a supernatant of a culture medium during induction of stem cells, a supernatant of a culture medium of stem cells, or a secretion product of stem cells. The therapeutic or preventive agent for wounds of cartilage tissue may further comprise hyaluronic acid.

[0027] According to an aspect of the present invention, there is provided a therapeutic or preventive agent for joint pain, comprising stem cells, a stem cell extract, a stem cell medium, a culture supernatant during stem cell induction, a culture supernatant of stem cells, or a secretion product of stem cells. The therapeutic or preventive agent for joint pain may further comprise hyaluronic acid.

[0028] According to an aspect of the present invention, there is provided an anti-aging agent comprising stem cells, a stem cell extract, a stem cell culture medium, a culture supernatant during stem cell induction, a stem cell culture supernatant, or a stem cell secretion product.

[0029] According to an aspect of the present invention, there is provided a rejuvenation agent comprising stem cells, a stem cell extract, a stem cell culture medium, a supernatant of a culture medium during stem cell induction, a supernatant of a culture medium of stem cells, or a secretion product of stem cells.

[0030] According to an aspect of the present invention, there is provided an agent for amplifying expression of a rejuvenation factor or a rejuvenation-related factor, comprising stem cells, a stem cell extract, a stem cell culture medium, a culture medium supernatant during stem cell induction, a stem cell culture medium supernatant, or a stem cell secretion product.

[0031] According to an aspect of the present invention, there is provided an expression amplification agent for at least any one of GDF11, MANF, TIMP2, SPARCL1, THBS4, NAMPT, and NANOG, comprising stem cells, a stem cell extract, stem cell culture medium, a culture medium supernatant during stem cell induction, a stem cell culture medium supernatant, or a stem cell secretion product.

[0032] According to an aspect of the present invention, there is provided an agent for inhibiting the expression of a senescence marker, comprising stem cells, a stem cell extract, a stem cell culture medium, a culture supernatant during stem cell induction, a culture supernatant of a stem cell culture medium, or a secretion product of a stem cell.

[0033] According to an aspect of the present invention, there is provided an agent for inhibiting the expression of at least one of p53 and p16, comprising stem cells, a stem cell extract, stem cell culture medium, a culture supernatant during stem cell induction, a stem cell culture supernatant, or a stem cell secretion product.

[0034] According to an aspect of the present invention, there is provided a therapeutic or preventive agent for lifestyle-related diseases, comprising stem cells, a stem cell extract, a stem cell culture medium, a culture supernatant during stem cell induction, a stem cell culture supernatant, or a stem cell secretion product.

[0035] According to an aspect of the present invention, there is provided a therapeutic or preventive agent for adult diseases, comprising stem cells, a stem cell extract, a stem cell culture medium, a culture supernatant during stem cell induction, a stem cell culture supernatant, or a stem cell secretion product.

[0036] According to an aspect of the present invention, there is provided an agent for improving the survival rate of chondrocytes, comprising stem cells, an extract of stem cells, a medium for stem cells, a supernatant of a culture medium during induction of stem cells, a supernatant of a culture medium for stem cells, or a secretion product of stem cells. The agent for improving the survival rate of chondrocytes may further comprise hyaluronic acid.

[0037] According to an aspect of the present invention, there is provided an analgesic agent comprising stem cells, an extract of stem cells, a medium for stem cells, a supernatant of a culture medium during induction of stem cells, a supernatant of a culture medium for stem cells, or a secretion product of stem cells. The analgesic agent may further comprise hyaluronic acid.

[0038] According to an aspect of the present invention, there is provided a cartilage regenerating agent comprising stem cells, a stem cell extract, a stem cell medium, a culture supernatant during stem cell induction, a culture supernatant of stem cells, or a secretion product of stem cells. The cartilage regenerating agent may further comprise hyaluronic acid.

[0039] According to an aspect of the present invention, there is provided a therapeutic, ameliorating or prophylactic agent for joints, joint diseases or arthritis, comprising stem cells, a stem cell extract, a stem cell medium, a culture supernatant during stem cell induction, a culture supernatant of stem cells, or a secretion product of stem cells. The therapeutic, ameliorating or prophylactic agent for joints, joint diseases or arthritis may further comprise hyaluronic acid.

[0040] According to an aspect of the present invention, there is provided an agent for suppressing or alleviating inflammation, comprising stem cells, a stem cell extract, a stem cell medium, a culture supernatant during stem cell induction, a culture supernatant of stem cells, or a secretion product of stem cells. The agent for suppressing or alleviating inflammation may further comprise hyaluronic acid.

[0041] According to an aspect of the present invention, there is provided an agent for promoting the production of an extracellular matrix, comprising stem cells, an extract of stem cells, a medium for stem cells, a supernatant of a culture medium during induction of stem cells, a supernatant of a culture medium for stem cells, or a secretion product of stem cells. The agent for promoting the production of an extracellular matrix may further comprise hyaluronic acid.

[0042] According to an aspect of the present invention, there is provided a telomere elongation agent or telomere shortening inhibitor comprising stem cells, a stem cell extract, stem cell culture medium, a culture medium supernatant during stem cell induction, a stem cell culture medium supernatant, or a stem cell secretion product.

[0043] According to an aspect of the present invention, there is provided a cell initialization agent, a cell de-differentiation agent, or a cell reprogramming agent, comprising stem cells, a stem cell extract, a stem cell culture medium, a culture medium supernatant during stem cell induction, a stem cell culture medium supernatant, or a stem cell secretion product.

[0044] According to an aspect of the present invention, there is provided the use of stem cells, stem cell extracts, stem cell culture medium, supernatant of culture medium during stem cell induction, supernatant of culture medium of stem cells, or secretion products of stem cells in the manufacture of a lipolytic agent.

[0045] According to an aspect of the present invention, there is provided the use of stem cells, stem cell extracts, stem cell culture medium, supernatant of culture medium during stem cell induction, supernatant of culture medium of stem cells, or secretion product of stem cells in the manufacture of an anti-obesity drug.

[0046] According to an aspect of the present invention, there is provided the use of stem cells, stem cell extracts, stem cell culture medium, supernatant of culture medium during stem cell induction, supernatant of stem cell culture medium, or stem cell secretion product in the manufacture of an obesity-improving drug.

[0047] According to an aspect of the present invention, there is provided the use of stem cells, stem cell extracts, stem cell culture medium, supernatant of culture medium during stem cell induction, supernatant of culture medium of stem cells, or secretion products of stem cells in the manufacture of a lipolysis promoter.

[0048] According to an aspect of the present invention, there is provided the use of stem cells, stem cell extracts, stem cell culture medium, supernatant of culture medium during stem cell induction, supernatant of culture medium of stem cells, or secretion product of stem cells in the manufacture of a dietary agent.

[0049] According to an aspect of the present invention, there is provided use of stem cells, stem cell extracts, stem cell culture medium, supernatant of culture medium during stem cell induction, supernatant of culture medium of stem cells, or secretion product of stem cells in the manufacture of a medicament for treating or preventing diabetes.

[0050] According to an aspect of the present invention, there is provided the use of stem cells, stem cell extracts, stem cell culture medium, supernatant of culture medium during stem cell induction, supernatant of stem cell culture medium, or stem cell secretions in the manufacture of an agent for promoting differentiation induction from myoblasts to myotubes.

[0051] According to an aspect of the present invention, there is provided the use of stem cells, stem cell extracts, stem cell culture medium, supernatant of culture medium during stem cell induction, supernatant of culture medium of stem cells, or secretion product of stem cells in the manufacture of an agent for promoting muscle formation.

[0052] According to an aspect of the present invention, there is provided use of stem cells, a stem cell extract, a stem cell culture medium, a culture supernatant during stem cell induction, a stem cell culture supernatant, or a stem cell secretion product in the manufacture of an agent for maintaining muscle mass.

[0053] According to an aspect of the present invention, there is provided the use of stem cells, stem cell extracts, stem cell culture medium, supernatant of culture medium during stem cell induction, supernatant of culture medium of stem cells, or secretion product of stem cells in the manufacture of an agent for increasing muscle mass.

[0054] According to an aspect of the present invention, there is provided the use of stem cells, stem cell extracts, stem cell culture medium, supernatant of culture medium during stem cell induction, supernatant of culture medium of stem cells, or secretion product of stem cells in the manufacture of a prostacyclin production promoter.

[0055] According to an aspect of the present invention, there is provided the use of stem cells, stem cell extracts, stem cell culture medium, supernatant of culture medium during stem cell induction, supernatant of culture medium of stem cells, or secretions of stem cells in the manufacture of an adenylate cyclase activator.

[0056] According to an aspect of the present invention, there is provided the use of stem cells, stem cell extracts, stem cell culture medium, supernatant of culture medium during stem cell induction, supernatant of culture medium of stem cells, or secretion products of stem cells in the manufacture of a cyclic adenosine monophosphate production promoter.

[0057] According to an aspect of the present invention, there is provided the use of stem cells, stem cell extracts, stem cell culture medium, supernatant of culture medium during stem cell induction, supernatant of culture medium of stem cells, or secretion product of stem cells in the manufacture of a platelet aggregation inhibitor.

[0058] According to an aspect of the present invention, there is provided the use of stem cells, stem cell extracts, stem cell culture medium, supernatant of culture medium during stem cell induction, supernatant of culture medium of stem cells, or secretion product of stem cells in the manufacture of a vasodilator.

[0059] According to an aspect of the present invention, there is provided use of stem cells, stem cell extracts, stem cell culture medium, supernatant of culture medium during stem cell induction, supernatant of culture medium of stem cells, or secretion product of stem cells in the manufacture of a medicament for treating or preventing hypertension.

[0060] According to an aspect of the present invention, there is provided use of stem cells, stem cell extracts, stem cell culture medium, supernatant of culture medium during stem cell induction, supernatant of stem cell culture medium, or stem cell secretion product in the manufacture of a drug for the treatment or prevention of arteriosclerosis.

[0061] According to an aspect of the present invention, there is provided the use of stem cells, stem cell extracts, stem cell culture medium, supernatant of culture medium during stem cell induction, supernatant of culture medium of stem cells, or secretion product of stem cells in the manufacture of an inhibitor of vascular endothelial cell proliferation.

[0062] According to an aspect of the present invention, there is provided the use of stem cells, stem cell extracts, stem cell culture medium, supernatant of culture medium during stem cell induction, supernatant of stem cell culture medium, or stem cell secretions in the manufacture of an agent for promoting hyaluronic acid production in chondrocytes.

[0063] According to an aspect of the present invention, there is provided the use of stem cells, stem cell extracts, stem cell culture medium, supernatant of culture medium during stem cell induction, supernatant of stem cell culture medium, or stem cell secretion product in the manufacture of a chondrocyte protective drug or proliferation promoter.

[0064] According to an aspect of the present invention, there is provided use of stem cells, stem cell extracts, stem cell culture medium, supernatant of culture medium during stem cell induction, supernatant of culture medium of stem cells, or secretion product of stem cells in the manufacture of a medicament for treating or preventing wounds in cartilage tissue.

[0065] According to an aspect of the present invention, there is provided use of stem cells, stem cell extracts, stem cell culture medium, supernatant of culture medium during stem cell induction, supernatant of culture medium of stem cells, or secretion product of stem cells in the manufacture of a medicament for treating or preventing joint pain.

[0066] According to an aspect of the present invention, there is provided use of stem cells, a stem cell extract, a stem cell culture medium, a culture supernatant during stem cell induction, a stem cell culture supernatant, or a stem cell secretion product in the manufacture of an anti-aging agent.

[0067] According to an aspect of the present invention, there is provided the use of stem cells, stem cell extracts, stem cell culture medium, supernatant of culture medium during stem cell induction, supernatant of culture medium of stem cells, or secretion product of stem cells in the manufacture of a rejuvenation agent.

[0068] According to an aspect of the present invention, there is provided the use of stem cells, stem cell extracts, stem cell culture medium, supernatant of culture medium during stem cell induction, supernatant of stem cell culture medium, or stem cell secretions in the manufacture of an agent for amplifying expression of a rejuvenation factor or rejuvenation-related factor.

[0069] According to an aspect of the present invention, there is provided the use of stem cells, stem cell extracts, stem cell culture medium, supernatant of culture medium during stem cell induction, supernatant of stem cell culture medium, or stem cell secretions in the manufacture of an expression amplification agent for at least any of GDF11, MANF, TIMP2, SPARCL1, THBS4, NAMPT, and NANOG.

[0070] According to an aspect of the present invention, there is provided the use of stem cells, stem cell extracts, stem cell culture medium, supernatant of culture medium during stem cell induction, supernatant of stem cell culture medium, or stem cell secretion product in the manufacture of an inhibitor of expression of a senescence marker.

[0071] According to an aspect of the present invention, there is provided the use of stem cells, stem cell extracts, stem cell culture medium, supernatant of culture medium during stem cell induction, supernatant of stem cell culture medium, or stem cell secretions in the manufacture of an inhibitor of the expression of at least one of p53 and p16.

[0072] According to an aspect of the present invention, there is provided use of stem cells, stem cell extracts, stem cell culture medium, supernatant of culture medium during stem cell induction, supernatant of stem cell culture medium, or stem cell secretion product in the manufacture of a drug for the treatment or prevention of lifestyle-related diseases.

[0073] According to an aspect of the present invention, there is provided use of stem cells, a stem cell extract, a stem cell culture medium, a culture supernatant during stem cell induction, a stem cell culture supernatant, or a stem cell secretion product in the manufacture of a drug for the treatment or prevention of an adult disease.

[0074] According to an aspect of the present invention, there is provided the use of stem cells, a stem cell extract, a stem cell culture medium, a supernatant of a culture medium during stem cell induction, a supernatant of a stem cell culture medium, or a stem cell secretion product in the manufacture of an agent for improving the survival rate of chondrocytes.

[0075] According to an aspect of the present invention, there is provided use of stem cells, stem cell extracts, stem cell culture medium, supernatant of culture medium during stem cell induction, supernatant of stem cell culture medium, or supernatant of stem cell secretion in the manufacture of a painkiller.

[0076] According to an aspect of the present invention, there is provided use of stem cells, stem cell extracts, stem cell culture medium, supernatant of culture medium during stem cell induction, supernatant of culture medium of stem cells, or secretion product of stem cells in the manufacture of a cartilage regeneration agent.

[0077] According to an aspect of the present invention, there is provided use of stem cells, stem cell extracts, stem cell culture medium, supernatant of culture medium during stem cell induction, supernatant of culture medium of stem cells, or secretion product of stem cells in the manufacture of a medicament for the treatment, amelioration, or prevention of a joint, a joint disease, or arthritis.

[0078] According to an aspect of the present invention, there is provided the use of stem cells, stem cell extracts, stem cell culture medium, supernatant of culture medium during stem cell induction, supernatant of culture medium of stem cells, or secretion product of stem cells in the manufacture of an agent for suppressing or alleviating inflammation.

[0079] According to an aspect of the present invention, there is provided the use of stem cells, a stem cell extract, a stem cell culture medium, a culture medium supernatant during stem cell induction, a stem cell culture medium supernatant, or a stem cell secretion product in the manufacture of an agent for promoting extracellular matrix production.

[0080] According to an aspect of the present invention, there is provided the use of stem cells, stem cell extracts, stem cell culture medium, supernatant of culture medium during stem cell induction, supernatant of stem cell culture medium, or stem cell secretion product in the manufacture of a telomere elongation agent or a telomere shortening inhibitor.

[0081] According to an aspect of the present invention, there is provided the use of a stem cell, a stem cell extract, a stem cell culture medium, a supernatant of a culture medium during stem cell induction, a supernatant of a culture medium of a stem cell, or a secretion product of a stem cell in the manufacture of a cell initialization agent, a cell de-differentiation agent, or a cell reprogramming agent.

[0082] According to an aspect of the present invention, there is provided a use of stem cells, a stem cell extract, a stem cell culture medium, a supernatant of a culture medium during stem cell induction, a supernatant of a culture medium of stem cells, or a secretion product of stem cells for lipolysis therapy.

[0083] According to an aspect of the present invention, there is provided a use of stem cells, stem cell extracts, stem cell culture medium, supernatant of culture medium during stem cell induction, supernatant of culture medium of stem cells, or secretion product of stem cells for anti-obesity treatment.

[0084] According to an aspect of the present invention, there is provided use of stem cells, stem cell extracts, stem cell culture medium, supernatant of culture medium during stem cell induction, supernatant of culture medium of stem cells, or secretion product of stem cells for the treatment of improving obesity.

[0085] According to an aspect of the present invention, there is provided use of stem cells, stem cell extracts, stem cell culture medium, supernatant of culture medium during stem cell induction, supernatant of culture medium of stem cells, or secretion product of stem cells for lipolysis promoting therapy.

[0086] According to an aspect of the present invention, there is provided the use of stem cells, stem cell extracts, stem cell culture medium, supernatant of culture medium during stem cell induction, supernatant of culture medium of stem cells, or secretions of stem cells for dietary therapy.

[0087] According to an aspect of the present invention, there is provided use of stem cells, a stem cell extract, a stem cell culture medium, a culture supernatant during stem cell induction, a stem cell culture supernatant, or a stem cell secretion product for the treatment or prevention of diabetes.

[0088] According to an aspect of the present invention, there is provided use of stem cells, a stem cell extract, a stem cell culture medium, a supernatant of a culture medium during stem cell induction, a supernatant of a stem cell culture medium, or a secretion product of stem cells for treatment promoting the induction of differentiation of myoblasts into myotubes.

[0089] According to an aspect of the present invention, there is provided use of stem cells, a stem cell extract, a stem cell culture medium, a supernatant of a culture medium during stem cell induction, a supernatant of a culture medium of stem cells, or a secretion product of stem cells for muscle formation promotion therapy.

[0090] According to an aspect of the present invention, there is provided a use of stem cells, a stem cell extract, a stem cell culture medium, a supernatant of a culture medium during stem cell induction, a supernatant of a culture medium of stem cells, or a secretion product of stem cells for muscle mass maintenance therapy.

[0091] According to an aspect of the present invention, there is provided a use of stem cells, a stem cell extract, a stem cell culture medium, a supernatant of a culture medium during stem cell induction, a supernatant of a culture medium of stem cells, or a secretion product of stem cells for the treatment of increasing muscle mass.

[0092] According to an aspect of the present invention, there is provided use of stem cells, stem cell extracts, stem cell culture medium, supernatant of culture medium during stem cell induction, supernatant of culture medium of stem cells, or secretion product of stem cells for prostacyclin production promoting therapy.

[0093] According to an aspect of the present invention, there is provided a use of stem cells, a stem cell extract, a stem cell culture medium, a supernatant of a culture medium during stem cell induction, a supernatant of a culture medium of stem cells, or a secretion product of stem cells for treatment of adenylate cyclase activity.

[0094] According to an aspect of the present invention, there is provided use of stem cells, stem cell extracts, stem cell culture medium, supernatant of culture medium during stem cell induction, supernatant of culture medium of stem cells, or secretion product of stem cells for treatment promoting cyclic adenosine monophosphate production.

[0095] According to an aspect of the present invention, there is provided use of stem cells, stem cell extracts, stem cell culture medium, supernatant of culture medium during stem cell induction, supernatant of culture medium of stem cells, or secretion product of stem cells for platelet aggregation inhibition therapy.

[0096] According to an aspect of the present invention, there is provided use of stem cells, stem cell extracts, stem cell culture medium, supernatant of culture medium during stem cell induction, supernatant of culture medium of stem cells, or secretion products of stem cells for vasodilatory therapy.

[0097] According to an aspect of the present invention, there is provided use of stem cells, a stem cell extract, a stem cell culture medium, a supernatant of a culture medium during stem cell induction, a supernatant of a culture medium of stem cells, or a secretion product of stem cells as a drug for the treatment or prevention of hypertension.

[0098] According to an aspect of the present invention, there is provided use of stem cells, a stem cell extract, a stem cell culture medium, a culture supernatant during stem cell induction, a stem cell culture supernatant, or a stem cell secretion product for the treatment or prevention of arteriosclerosis.

[0099] According to an aspect of the present invention, there is provided use of stem cells, stem cell extracts, stem cell culture medium, supernatant of culture medium during stem cell induction, supernatant of culture medium of stem cells, or secretion product of stem cells for the treatment of inhibiting vascular endothelial cell proliferation.

[0100] According to an aspect of the present invention, there is provided use of stem cells, stem cell extracts, stem cell culture medium, supernatant of culture medium during stem cell induction, supernatant of culture medium of stem cells, or secretion product of stem cells for treatment of promoting hyaluronic acid production in chondrocytes.

[0101] According to an aspect of the present invention, there is provided use of stem cells, a stem cell extract, a stem cell culture medium, a supernatant of a culture medium during stem cell induction, a supernatant of a culture medium of stem cells, or a secretion product of stem cells for treatment for protecting or promoting the proliferation of chondrocytes.

[0102] According to an aspect of the present invention, there is provided use of stem cells, a stem cell extract, a stem cell culture medium, a supernatant of a culture medium during stem cell induction, a supernatant of a culture medium of stem cells, or a secretion product of stem cells for the treatment or prevention of wounds in cartilage tissue.

[0103] According to an aspect of the present invention, there is provided use of stem cells, stem cell extracts, stem cell culture medium, supernatant of culture medium during stem cell induction, supernatant of culture medium of stem cells, or secretion product of stem cells for the treatment or prevention of joint pain.

[0104] According to an aspect of the present invention, there is provided a use of stem cells, a stem cell extract, a stem cell culture medium, a supernatant of a culture medium during stem cell induction, a supernatant of a culture medium of stem cells, or a secretion product of stem cells for anti-aging treatment.

[0105] According to an aspect of the present invention, there is provided use of stem cells, stem cell extracts, stem cell culture medium, supernatant of culture medium during stem cell induction, supernatant of culture medium of stem cells, or secretion product of stem cells for rejuvenation therapy.

[0106] According to an aspect of the present invention, there is provided the use of stem cells, stem cell extracts, stem cell culture medium, supernatant of culture medium during stem cell induction, supernatant of culture medium of stem cells, or secretions of stem cells for amplifying the expression of rejuvenation factors or rejuvenation-related factors.

[0107] According to an aspect of the present invention, there is provided use of stem cells, stem cell extracts, stem cell culture medium, supernatant of culture medium during stem cell induction, supernatant of culture medium of stem cells, or secretions of stem cells for amplifying expression of at least any of GDF11, MANF, TIMP2, SPARCL1, THBS4, NAMPT, and NANOG.

[0108] According to an aspect of the present invention, there is provided use of stem cells, stem cell extracts, stem cell culture medium, supernatant of culture medium during stem cell induction, supernatant of culture medium of stem cells, or secretion products of stem cells for suppressing expression of senescence markers.

[0109] According to an aspect of the present invention, there is provided use of stem cells, stem cell extracts, stem cell culture medium, supernatant of culture medium during stem cell induction, supernatant of culture medium of stem cells, or secretion products of stem cells for suppressing expression of at least one of p53 and p16.

[0110] According to an aspect of the present invention, there is provided use of stem cells, a stem cell extract, a stem cell culture medium, a culture supernatant during stem cell induction, a stem cell culture supernatant, or a stem cell secretion product for the treatment or prevention of lifestyle-related diseases.

[0111] According to an aspect of the present invention, there is provided use of stem cells, a stem cell extract, a stem cell culture medium, a culture supernatant during stem cell induction, a stem cell culture supernatant, or a stem cell secretion product for the treatment or prevention of an adult disease.

[0112] According to an aspect of the present invention, there is provided use of stem cells, a stem cell extract, a stem cell culture medium, a supernatant of a culture medium during stem cell induction, a supernatant of a culture medium of stem cells, or a secretion product of stem cells for improving the survival rate of chondrocytes.

[0113] According to an aspect of the present invention, there is provided a use of stem cells, a stem cell extract, a stem cell culture medium, a supernatant of a culture medium during stem cell induction, a supernatant of a stem cell culture medium, or a supernatant of a stem cell secretion product for analgesia.

[0114] According to an aspect of the present invention, there is provided use of stem cells, a stem cell extract, a stem cell culture medium, a supernatant of a culture medium during stem cell induction, a supernatant of a culture medium of stem cells, or a secretion product of stem cells for cartilage regeneration.

[0115] According to an aspect of the present invention, there is provided use of stem cells, a stem cell extract, a stem cell culture medium, a supernatant of a culture medium during stem cell induction, a supernatant of a culture medium of stem cells, or a secretion product of stem cells for the treatment, amelioration, or prevention of a joint, a joint disease, or arthritis.

[0116] According to an aspect of the present invention, there is provided use of stem cells, a stem cell extract, a stem cell culture medium, a supernatant of a culture medium during stem cell induction, a supernatant of a culture medium of stem cells, or a secretion product of stem cells for suppressing or alleviating inflammation.

[0117] According to an aspect of the present invention, there is provided use of stem cells, a stem cell extract, a stem cell culture medium, a supernatant of a culture medium during stem cell induction, a supernatant of a culture medium of stem cells, or a secretion product of stem cells for promoting production of extracellular matrix.

[0118] According to an aspect of the present invention, there is provided use of stem cells, stem cell extracts, stem cell culture medium, supernatant of culture medium during stem cell induction, supernatant of culture medium of stem cells, or secretion products of stem cells for telomere elongation or inhibition of telomere shortening.

[0119] According to an aspect of the present invention, there is provided a use of a stem cell, a stem cell extract, a stem cell medium, a supernatant of a culture medium during stem cell induction, a supernatant of a culture medium of a stem cell, or a secretion product of a stem cell for reprogramming a cell, dedifferentiating a cell, or reprogramming a cell.

[0120] According to an aspect of the present invention, there is provided a method for lipolysis treatment, comprising administering a lipolysis agent comprising stem cells, a stem cell extract, a stem cell medium, a culture supernatant during stem cell induction, a culture supernatant of stem cells, or a secretion product of stem cells to a subject, the subject being a human or a non-human animal.

[0121] According to an aspect of the present invention, there is provided a method for treating obesity, comprising administering to a subject an anti-obesity drug comprising stem cells, a stem cell extract, stem cell culture medium, a culture medium supernatant during stem cell induction, a stem cell culture medium supernatant, or a stem cell secretion product.

[0122] According to an aspect of the present invention, there is provided a therapeutic method for improving obesity, which comprises administering to a subject an obesity-improving drug containing stem cells, a stem cell extract, stem cell culture medium, a culture medium supernatant during stem cell induction, a stem cell culture medium supernatant, or a stem cell secretion product.

[0123] According to an aspect of the present invention, there is provided a method for promoting lipolysis, comprising administering to a subject a lipolysis promoter comprising stem cells, a stem cell extract, a stem cell culture medium, a culture medium supernatant during stem cell induction, a stem cell culture medium supernatant, or a stem cell secretion product.

[0124] According to an aspect of the present invention, there is provided a diet treatment method comprising administering to a subject a diet agent containing stem cells, a stem cell extract, stem cell culture medium, a culture medium supernatant during stem cell induction, a stem cell culture medium supernatant, or a stem cell secretion product.

[0125] According to an aspect of the present invention, there is provided a method for treating or preventing diabetes, comprising administering to a subject a drug for treating or preventing diabetes, the drug comprising stem cells, a stem cell extract, a stem cell culture medium, a culture medium supernatant during stem cell induction, a stem cell culture medium supernatant, or a stem cell secretion product.

[0126] According to an aspect of the present invention, there is provided a therapeutic method for promoting differentiation of myoblasts into myotubes, comprising administering to a subject an agent for promoting the induction of differentiation of myoblasts into myotubes, the agent comprising stem cells, a stem cell extract, a stem cell medium, a culture medium supernatant during stem cell induction, a stem cell culture medium supernatant, or a stem cell secretion product.

[0127] According to an aspect of the present invention, there is provided a therapeutic method for promoting muscle formation, comprising administering to a subject a muscle formation promoter comprising stem cells, a stem cell extract, a stem cell culture medium, a culture medium supernatant during stem cell induction, a stem cell culture medium supernatant, or a stem cell secretion product.

[0128] According to an aspect of the present invention, there is provided a therapeutic method for maintaining muscle mass, comprising administering to a subject a muscle mass maintenance agent comprising stem cells, a stem cell extract, a stem cell culture medium, a culture medium supernatant during stem cell induction, a stem cell culture medium supernatant, or a stem cell secretion product.

[0129] According to an aspect of the present invention, there is provided a therapeutic method for increasing muscle mass, comprising administering to a subject an agent for increasing muscle mass, the agent comprising stem cells, a stem cell extract, a stem cell culture medium, a culture medium supernatant during stem cell induction, a stem cell culture medium supernatant, or a stem cell secretion product.

[0130] According to an aspect of the present invention, there is provided a therapeutic method for promoting prostacyclin production, comprising administering to a subject a prostacyclin production promoter comprising stem cells, a stem cell extract, a stem cell culture medium, a culture medium supernatant during stem cell induction, a stem cell culture medium supernatant, or a stem cell secretion product.

[0131] According to an aspect of the present invention, there is provided a therapeutic method for activating adenylate cyclase, comprising administering to a subject an adenylate cyclase activator comprising stem cells, a stem cell extract, stem cell culture medium, the supernatant of a culture medium during stem cell induction, the supernatant of a stem cell culture medium, or a secretion product of a stem cell.

[0132] According to an aspect of the present invention, there is provided a therapeutic method for promoting cyclic adenosine monophosphate production, comprising administering to a subject a cyclic adenosine monophosphate production promoter comprising stem cells, a stem cell extract, a stem cell culture medium, a culture medium supernatant during stem cell induction, a stem cell culture medium supernatant, or a stem cell secretion product.

[0133] According to an aspect of the present invention, there is provided a platelet aggregation inhibitor treatment method comprising administering to a subject a platelet aggregation inhibitor comprising stem cells, a stem cell extract, a stem cell culture medium, a culture medium supernatant during stem cell induction, a stem cell culture medium supernatant, or a stem cell secretion product.

[0134] According to an aspect of the present invention, there is provided a method for vasodilatory treatment, comprising administering to a subject a vasodilator comprising stem cells, a stem cell extract, a stem cell culture medium, a culture medium supernatant during stem cell induction, a stem cell culture medium supernatant, or a stem cell secretion product.

[0135] According to an aspect of the present invention, there is provided a method for treating or preventing hypertension, comprising administering to a subject a drug for treating or preventing hypertension, the drug comprising stem cells, a stem cell extract, a stem cell culture medium, a culture medium supernatant during stem cell induction, a stem cell culture medium supernatant, or a stem cell secretion product.

[0136] According to an aspect of the present invention, there is provided a method for treating or preventing arteriosclerosis, comprising administering to a subject an agent for treating or preventing arteriosclerosis, the agent comprising stem cells, a stem cell extract, stem cell culture medium, a culture medium supernatant during stem cell induction, a culture medium supernatant of stem cells, or a secretion product of stem cells.

[0137] According to an aspect of the present invention, there is provided a vascular endothelial cell proliferation inhibitor treatment, which comprises administering to a subject a vascular endothelial cell proliferation inhibitor comprising stem cells, a stem cell extract, a stem cell culture medium, a culture medium supernatant during stem cell induction, a stem cell culture medium supernatant, or a stem cell secretion product.

[0138] According to an aspect of the present invention, there is provided a therapeutic method for promoting hyaluronic acid production in chondrocytes, comprising administering to a subject a promoter for promoting hyaluronic acid production in chondrocytes, the promoter comprising stem cells, a stem cell extract, a stem cell culture medium, a culture medium supernatant during stem cell induction, a stem cell culture medium supernatant, or a stem cell secretion product.

[0139] According to an aspect of the present invention, there is provided a therapeutic method for protecting or promoting the proliferation of chondrocytes, which comprises administering to a subject a protective agent or proliferation promoter for chondrocytes, which comprises stem cells, a stem cell extract, a stem cell culture medium, a culture medium supernatant during stem cell induction, a culture medium supernatant for stem cells, or a secretion product of stem cells.

[0140] According to an aspect of the present invention, there is provided a method for treating or preventing wounds in cartilage tissue, comprising administering to a subject a therapeutic or preventive agent for wounds in cartilage tissue, the therapeutic or preventive agent comprising stem cells, a stem cell extract, a stem cell culture medium, a culture medium supernatant during stem cell induction, a stem cell culture medium supernatant, or a stem cell secretion product.

[0141] According to an aspect of the present invention, there is provided a method for treating or preventing joint pain, comprising administering to a subject a therapeutic or preventive agent for joint pain, the therapeutic or preventive agent comprising stem cells, a stem cell extract, a stem cell culture medium, a culture medium supernatant during stem cell induction, a stem cell culture medium supernatant, or a stem cell secretion product.

[0142] According to an aspect of the present invention, there is provided an anti-aging treatment method comprising administering to a subject an anti-aging agent comprising stem cells, a stem cell extract, a stem cell culture medium, a culture supernatant during stem cell induction, a stem cell culture supernatant, or a stem cell secretion product.

[0143] According to an aspect of the present invention, there is provided a rejuvenation method comprising administering to a subject a rejuvenation agent comprising stem cells, a stem cell extract, a stem cell culture medium, a culture medium supernatant during stem cell induction, a stem cell culture medium supernatant, or a stem cell secretion product.

[0144] According to an aspect of the present invention, there is provided a method for amplifying expression of a rejuvenation factor or a rejuvenation-associated factor, comprising administering to a subject a rejuvenation factor or a rejuvenation-associated factor, including stem cells, a stem cell extract, a stem cell culture medium, a culture medium supernatant during stem cell induction, a stem cell culture medium supernatant, or a stem cell secretion product.

[0145] According to an aspect of the present invention, there is provided a method for amplifying the expression of at least one of GDF11, MANF, TIMP2, SPARCL1, THBS4, NAMPT, and NANOG, comprising administering to a subject an expression amplifying agent for at least one of GDF11, MANF, TIMP2, SPARCL1, THBS4, NAMPT, and NANOG, which comprises stem cells, a stem cell extract, stem cell culture medium, the supernatant of a culture medium during stem cell induction, the supernatant of a stem cell culture medium, or a secretion product of a stem cell.

[0146] According to an aspect of the present invention, there is provided a method for inhibiting expression of a senescence marker, comprising administering to a subject an expression inhibitor of a senescence marker, the inhibitor comprising stem cells, a stem cell extract, a stem cell culture medium, a culture medium supernatant during stem cell induction, a stem cell culture medium supernatant, or a stem cell secretion product.

[0147] According to an aspect of the present invention, there is provided a method for inhibiting the expression of at least one of p53 and p16, comprising administering to a subject an expression inhibitor of at least one of p53 and p16, which comprises stem cells, a stem cell extract, a stem cell culture medium, a culture medium supernatant during stem cell induction, a stem cell culture medium supernatant, or a stem cell secretion product.

[0148] According to an aspect of the present invention, there is provided a method for treating lifestyle-related diseases, which comprises administering to a subject a therapeutic or preventive drug for lifestyle-related diseases, which comprises stem cells, a stem cell extract, stem cell culture medium, a culture medium supernatant during stem cell induction, a stem cell culture medium supernatant, or a stem cell secretion product.

[0149] According to an aspect of the present invention, there is provided a method for treating or preventing an adult disease, comprising administering to a subject a therapeutic or preventive drug for the adult disease, the therapeutic or preventive drug comprising stem cells, a stem cell extract, a stem cell medium, a culture supernatant during stem cell induction, a stem cell culture supernatant, or a stem cell secretion product.

[0150] According to an aspect of the present invention, there is provided a method for improving the viability of chondrocytes, comprising administering to a subject an agent for improving the viability of chondrocytes, which comprises stem cells, a stem cell extract, a stem cell culture medium, a culture medium supernatant during stem cell induction, a stem cell culture medium supernatant, or a stem cell secretion product.

[0151] According to an aspect of the present invention, there is provided a method for relieving pain, comprising administering to a subject a pain relieving agent comprising stem cells, a stem cell extract, a stem cell culture medium, a culture supernatant during stem cell induction, a culture supernatant of stem cells, or a secretion product of stem cells.

[0152] According to an aspect of the present invention, there is provided a method for regenerating chondrocytes, comprising administering to a subject a cartilage regenerating agent comprising stem cells, a stem cell extract, a stem cell culture medium, a culture medium supernatant during stem cell induction, a stem cell culture medium supernatant, or a stem cell secretion product.

[0153] According to an aspect of the present invention, there is provided a method for treating, ameliorating or preventing a joint, a joint disease or arthritis, the method comprising administering to a subject a drug for treating, ameliorating or preventing a joint, a joint disease or arthritis, the drug comprising stem cells, a stem cell extract, a stem cell medium, a culture medium supernatant during stem cell induction, a stem cell culture medium supernatant, or a stem cell secretion product.

[0154] According to an aspect of the present invention, there is provided a method for suppressing or alleviating inflammation, comprising administering to a subject an agent for suppressing or alleviating inflammation, the agent comprising stem cells, a stem cell extract, a stem cell culture medium, a culture medium supernatant during stem cell induction, a stem cell culture medium supernatant, or a stem cell secretion product.

[0155] According to an aspect of the present invention, there is provided a method for promoting the production of extracellular matrix, comprising administering to a subject an agent for promoting the production of extracellular matrix, the agent comprising stem cells, a stem cell extract, a stem cell culture medium, a culture medium supernatant during stem cell induction, a stem cell culture medium supernatant, or a stem cell secretion product.

[0156] According to an aspect of the present invention, there is provided a method for extending telomere or inhibiting telomere shortening, comprising administering to a subject a telomere extending agent or a telomere shortening inhibitor, the telomere extending agent or the telomere shortening inhibitor comprising stem cells, a stem cell extract, a stem cell culture medium, a culture medium supernatant during stem cell induction, a stem cell culture medium supernatant, or a stem cell secretion product.

[0157] According to an aspect of the present invention, there is provided a method for initializing a cell, a method for dedifferentiating a cell, or a method for reprogramming a cell, comprising administering to a subject a cell initializing agent, a cell dedifferentiating agent, or a cell reprogramming agent, the cell dedifferentiating agent comprising stem cells, a stem cell extract, a stem cell culture medium, a supernatant of a culture medium during stem cell induction, a supernatant of a culture medium of a stem cell, or a secretion product of a stem cell. Effect of the Invention

[0158] According to the present invention, it is possible to provide pharmaceutical compositions and cosmetic compositions that effectively utilize stem cells and stem cell culture media. [Brief description of the drawings]

[0159] [Figure 1] 3 is a photograph of fat cells according to Example 2. [Diagram 2] 3 is a photograph of fat cells according to Example 2. [Diagram 3] 3 is a photograph of fat cells according to Example 2. [Figure 4] 1 is a table showing the measurement results of free glycerol when 5% of a test substance according to Example 2 was added to a culture medium for adipocytes. [Diagram 5] 1 is a graph showing the results of measuring free glycerol when 5% of a test substance according to Example 2 was added to a culture medium for adipocytes. [Figure 6]1 is a table showing the results of measuring free glycerol when 20% of a test substance according to Example 2 was added to a culture medium for adipocytes. [Figure 7] 1 is a graph showing the results of measuring free glycerol when 20% of a test substance according to Example 2 was added to a culture medium for adipocytes. [Figure 8] 1 is a table showing the measurement results of free glycerol when isoproterenol according to Example 2 is added to a culture medium for adipocytes. [Figure 9] 1 is a graph showing the measurement results of free glycerol when isoproterenol according to Example 2 is added to a culture medium for adipocytes. [Figure 10] 1 is a photograph of cells stained with an anti-MHC antibody according to Example 3. [Figure 11] 1 is a photograph of cells stained with an anti-MHC antibody according to Example 3. [Figure 12] 1 is a photograph of cells stained with an anti-MHC antibody according to Example 3. [Figure 13] 1 is a table showing the results of measuring the differentiation index into myotubes when 5% of a test substance according to Example 3 was added to a culture medium for normal human skeletal myoblasts. [Figure 14] 1 is a graph showing the measurement results of the differentiation index into myotubes when 5% of the test substance in Example 3 was added to a culture medium of normal human skeletal myoblasts. [Figure 15] 1 is a table showing the results of measuring the differentiation index into myotubes when 20% of a test substance according to Example 3 was added to a culture medium for normal human skeletal myoblasts. [Figure 16] 1 is a graph showing the results of measuring the differentiation index into myotubes when 20% of the test substance in Example 3 was added to a culture medium for normal human skeletal myoblasts. [Figure 17] 13 is a table showing the measurement results of the differentiation index into myotubes when the insulin according to Example 3 was added to a culture medium for normal human skeletal myoblasts. [Figure 18]13 is a graph showing the measurement results of the differentiation index into myotubes when the insulin according to Example 3 was added to a culture medium of normal human skeletal myoblasts. [Figure 19] 1 is a table showing the results of measuring the amount of 6-keto prostaglandin F1α when 5% of a test substance according to Example 4 was added to a culture medium for normal human umbilical vein endothelial cells. [Figure 20] 1 is a graph showing the amount of 6-keto prostaglandin F1α when 5% of the test substance according to Example 4 was added to a culture medium for normal human umbilical vein endothelial cells. [Figure 21] 1 is a table showing the results of measuring the amount of 6-keto prostaglandin F1α when 20% of the test substance according to Example 4 was added to a culture medium for normal human umbilical vein endothelial cells. [Figure 22] 1 is a graph showing the amount of 6-keto prostaglandin F1α when 20% of the test substance according to Example 4 was added to a culture medium for normal human umbilical vein endothelial cells. [Figure 23] 1 is a table showing the measurement results of the amount of 6-ketoprostaglandin F1α when arachidonic acid according to Example 4 was added to a culture medium of normal human umbilical vein endothelial cells. [Figure 24] 1 is a graph showing the amount of 6-keto prostaglandin F1α when arachidonic acid according to Example 4 is added to a culture medium of normal human umbilical vein endothelial cells. [Diagram 25] 1 is a table showing the results of measuring the amount of hyaluronic acid produced when 5% of the test substance in Example 5 was added to a culture medium of human chondrocytes and the culture medium of human chondrocytes was cultured for 24 hours. [Figure 26] 13 is a graph showing the amount of hyaluronic acid produced when 5% of the test substance in Example 5 was added to a culture medium of human chondrocytes and the culture medium of human chondrocytes was cultured for 24 hours. [Figure 27] 1 is a table showing the results of measuring the amount of hyaluronic acid produced when 20% of the test substance in Example 5 was added to a culture medium of human chondrocytes and the culture medium of human chondrocytes was cultured for 24 hours. [Figure 28]13 is a graph showing the amount of hyaluronic acid produced when 20% of the test substance in Example 5 was added to a culture medium of human chondrocytes and the culture medium was cultured for 24 hours. [Figure 29] 13 is a table showing the results of measuring the amount of hyaluronic acid produced when N-acetylglucosamine according to Example 5 was added to a culture medium of human chondrocytes and the culture medium of human chondrocytes was cultured for 24 hours. [Diagram 30] 13 is a graph showing the amount of hyaluronic acid produced when N-acetylglucosamine according to Example 5 was added to a culture medium of human chondrocytes and the culture medium of human chondrocytes was cultured for 24 hours. [Diagram 31] 1 is a table showing the results of measuring the amount of hyaluronic acid produced when 5% of the test substance in Example 5 was added to a culture medium of human chondrocytes and the culture medium of human chondrocytes was cultured for 72 hours. [Diagram 32] 13 is a graph showing the amount of hyaluronic acid produced when 5% of the test substance in Example 5 was added to a culture medium of human chondrocytes and the culture medium of human chondrocytes was cultured for 72 hours. [Diagram 33] 1 is a table showing the results of measuring the amount of hyaluronic acid produced when 20% of the test substance in Example 5 was added to a culture medium of human chondrocytes and the culture medium of human chondrocytes was cultured for 72 hours. [Diagram 34] 13 is a graph showing the amount of hyaluronic acid produced when 20% of the test substance in Example 5 was added to a culture medium of human chondrocytes and the culture medium of human chondrocytes was cultured for 72 hours. [Diagram 35] 13 is a table showing the results of measuring the amount of hyaluronic acid produced when N-acetylglucosamine according to Example 5 was added to a culture medium of human chondrocytes and the culture medium of human chondrocytes was cultured for 72 hours. [Diagram 36] 13 is a graph showing the amount of hyaluronic acid produced when N-acetylglucosamine according to Example 5 was added to a culture medium of human chondrocytes and the culture medium of human chondrocytes was cultured for 72 hours. [Figure 37] 1 is a table showing the results of measuring the amount of hyaluronic acid produced when 5% of the test substance in Example 5 was added to a culture medium of human chondrocytes and the culture medium of human chondrocytes was cultured for 24 hours. [Figure 38] 13 is a graph showing the amount of hyaluronic acid produced when 5% of the test substance in Example 5 was added to a culture medium of human chondrocytes and the culture medium of human chondrocytes was cultured for 24 hours. [Figure 39] 1 is a table showing the results of measuring the amount of hyaluronic acid produced when 20% of the test substance in Example 5 was added to a culture medium of human chondrocytes and the culture medium of human chondrocytes was cultured for 24 hours. [Diagram 40] 1 is a graph showing the amount of hyaluronic acid produced when 20% of the test substance according to Example 5 is added to a culture solution of human chondrocytes and the culture solution of human chondrocytes is cultured for 24 hours. FIG. 2 is a graph showing the amount of hyaluronic acid produced when a culture solution of mouse chondrocytes is cultured for 72 hours. [Diagram 41] 1 is a table showing the results of measuring the amount of hyaluronic acid produced when 5% of the test substance in Example 5 was added to a culture medium of human chondrocytes and the culture medium of human chondrocytes was cultured for 72 hours. [Diagram 42] 13 is a graph showing the amount of hyaluronic acid produced when 5% of the test substance in Example 5 was added to a culture medium of human chondrocytes and the culture medium of human chondrocytes was cultured for 72 hours. [Diagram 43] 1 is a table showing the results of measuring the amount of hyaluronic acid produced when 20% of the test substance in Example 5 was added to a culture medium of human chondrocytes and the culture medium of human chondrocytes was cultured for 72 hours. [Diagram 44] 13 is a graph showing the amount of hyaluronic acid produced when 20% of the test substance in Example 5 was added to a culture medium of human chondrocytes and the culture medium of human chondrocytes was cultured for 72 hours. [Diagram 45] 13 is a table showing the results of measuring the survival rate of human chondrocytes when 5% of a test substance in Example 5 was added to a culture medium of human chondrocytes and the culture medium of human chondrocytes was cultured for 24 hours. [Figure 46] 13 is a graph showing the survival rate of human chondrocytes when 5% of a test substance according to Example 5 was added to a culture medium of human chondrocytes and the culture medium was cultured for 24 hours. [Figure 47]13 is a table showing the results of measuring the survival rate of human chondrocytes when 20% of a test substance in Example 5 was added to a culture medium of human chondrocytes and the culture medium of human chondrocytes was cultured for 24 hours. [Figure 48] 13 is a graph showing the survival rate of human chondrocytes when 20% of a test substance according to Example 5 was added to a culture medium of human chondrocytes and the culture medium was cultured for 24 hours. [Figure 49] 13 is a table showing the results of measuring the survival rate of human chondrocytes when N-acetylglucosamine according to Example 5 was added to a culture medium of human chondrocytes and the culture medium of human chondrocytes was cultured for 24 hours. [Figure 50] 13 is a graph showing the survival rate of human chondrocytes when N-acetylglucosamine according to Example 5 was added to a culture medium of human chondrocytes and the culture medium of human chondrocytes was cultured for 24 hours. [Figure 51] 13 is a table showing the results of measuring the survival rate of human chondrocytes when 5% of a test substance in Example 5 was added to a culture medium of human chondrocytes and the culture medium of human chondrocytes was cultured for 72 hours. [Figure 52] 13 is a graph showing the survival rate of human chondrocytes when 5% of a test substance according to Example 5 was added to a culture medium of human chondrocytes and the culture medium was cultured for 72 hours. [Figure 53] 13 is a table showing the results of measuring the survival rate of human chondrocytes when 20% of a test substance in Example 5 was added to a culture medium of human chondrocytes and the culture medium of human chondrocytes was cultured for 72 hours. [Figure 54] 13 is a graph showing the survival rate of human chondrocytes when 20% of a test substance according to Example 5 was added to a culture medium of human chondrocytes and the culture medium was cultured for 72 hours. [Figure 55] 13 is a table showing the results of measuring the survival rate of human chondrocytes when N-acetylglucosamine according to Example 5 was added to a culture medium of human chondrocytes and the culture medium of human chondrocytes was cultured for 72 hours. [Figure 56] 13 is a graph showing the survival rate of human chondrocytes when N-acetylglucosamine according to Example 5 was added to a culture medium of human chondrocytes and the culture medium of human chondrocytes was cultured for 72 hours. [Figure 57] 1 is a micrograph of cells according to a reference example. [Figure 58] 1 is a histogram showing the results of flow cytometry of cells according to a reference example. [Figure 59] 13 is a graph showing the left hind limb load ratio of arthropathy model rats according to Example 6. [Figure 60] 1 is a graph showing gene expression levels according to Example 8. [Figure 61] 1 is a graph showing gene expression levels according to Example 8. [Figure 62] 1 is a graph showing gene expression levels according to Example 8. [Figure 63] 1 is a graph showing gene expression levels according to Example 8. [Figure 64] 1 is a graph showing gene expression levels according to Example 8. [Figure 65] 1 is a graph showing gene expression levels according to Example 8. [Figure 66] 1 is a graph showing gene expression levels according to Example 8. [Figure 67] 1 is a graph showing gene expression levels according to Example 8. [Figure 68] 1 is a graph showing gene expression levels according to Example 8. [Figure 69] 1 is a graph showing gene expression levels according to Example 8. [Figure 70] 13 is a graph showing gene expression levels according to Example 9. [Figure 71] 13 is a graph showing gene expression levels according to Example 9. [Figure 72] 13 is a graph showing gene expression levels according to Example 9. [Figure 73] 1 is a graph showing gene expression levels according to Example 10. [Figure 74] 1 is a graph showing gene expression levels according to Example 10. [Figure 75] 1 is a graph showing gene expression levels according to Example 10. [Figure 76] 1 is a graph showing gene expression levels according to Example 10. [Figure 77] 1 is a graph showing gene expression levels according to Example 10. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0160] The following is a detailed description of the embodiments of the present invention. Note that the embodiments shown below are merely examples of devices and methods for embodying the technical idea of ​​the present invention, and the technical idea of ​​the present invention is not limited to the combination of components described below. The technical idea of ​​the present invention can be modified in various ways within the scope of the claims.

[0161] The pharmaceutical composition and / or cosmetic composition according to the embodiment includes stem cells, a stem cell extract, a stem cell culture medium, a supernatant of a culture medium during stem cell induction, a supernatant of a stem cell culture medium, or a secretion product of a stem cell. The pharmaceutical composition and / or cosmetic composition according to the embodiment may include a combination of at least two or more selected from a stem cell extract, a stem cell culture medium, a supernatant of a culture medium during stem cell induction, and a supernatant of a stem cell culture medium.

[0162] Examples of the pharmaceutical composition and / or cosmetic composition include lipolysis agents, anti-obesity agents, obesity improving agents, lipolysis promoters, diet agents, therapeutic or preventive agents for diabetes, agents for promoting differentiation of myoblasts into myotubes, muscle formation promoters, agents for maintaining muscle mass, agents for increasing muscle mass, prostacyclin production promoters, adenylate cyclase activators, cyclic adenosine monophosphate (cAMP) production promoters, platelet aggregation inhibitors, vasodilators, therapeutic or preventive agents for hypertension, therapeutic or preventive agents for arteriosclerosis, vascular endothelial cell proliferation inhibitors, agents for promoting hyaluronic acid production in chondrocytes, agents for promoting the proliferation of chondrocytes, agents for protecting chondrocytes, therapeutic or preventive agents for wounds in cartilage tissue, agents for promoting the production of extracellular matrix, therapeutic or preventive agents for joint pain, anti-aging agents, rejuvenating agents, rejuvenation agents, etc. the agent for amplifying expression of a factor, an agent for amplifying expression of a rejuvenation-related factor, an agent for amplifying expression of at least one of GDF11, MANF, TIMP2, SPARCL1, THBS4, NAMPT, NANOG, and TERT, an agent for suppressing expression of an aging marker, an agent for suppressing expression of a cellular aging marker, an agent for suppressing expression of p53, an agent for suppressing expression of p16, an inhibitor of the p53 pathway, an inhibitor of the p16 pathway, an inhibitor of the p19-p52 pathway, an inhibitor of the p16-Rb pathway, an agent for extending telomere length, an inhibitor of telomere shortening, an agent for initializing a cell, an agent for de-differentiating a cell, an agent for reprogramming a cell, a therapeutic or preventive agent for a lifestyle-related disease, a therapeutic or preventive agent for an adult disease, an agent for improving the survival rate of chondrocytes, an analgesic, a cartilage regenerating agent, and / or an agent for treating, improving, or preventing a joint, a joint disease, or arthritis.

[0163] The stem cells are, for example, pluripotent stem cells. The stem cells may be derived from humans or non-human animals. The pluripotent stem cells are, for example, iPS cells and embryonic stem cells (ES cells). The stem cell medium is, for example, a medium for stem cells that has not been used for culturing stem cells. The culture medium during stem cell induction is, for example, a culture medium used when somatic cells are reprogrammed into pluripotent stem cells such as iPS cells. The culture medium for stem cells is, for example, a culture medium used when maintaining and culturing stem cells. The stem cells, stem cell extracts, stem cell medium, supernatant of the culture medium during stem cell induction, supernatant of the culture medium for stem cells, or secretions of stem cells may be lyophilized. The stem cells, stem cell extracts, stem cell medium, supernatant of the culture medium during stem cell induction, supernatant of the culture medium for stem cells, or secretions of stem cells may be contained in a capsule such as a nanocapsule or an emulsion such as a nanoemulsion.

[0164] iPS cells are induced by introducing reprogramming factors such as OCT3 / 4, KLF4, c-MYC, and SOX2 into somatic cells, such as differentiated cells, such as blood cells, fibroblasts, and urine cells. Induction into iPS cells is sometimes called reprogramming, reprogramming, transformation, transdifferentiation or lineage reprogramming, and cell fate reprogramming. Pluripotent stem cells may be induced while being three-dimensionally cultured, such as in suspension culture. In the three-dimensional culture, a gel medium or a liquid medium may be used. In the three-dimensional culture, feeder cells may not be used. For example, the positivity rate of any one of TRA1-60, OCT3 / 4, SSEA3, SSEA4, TRA1-81, LIN28, and NANOG is 30% or more, 50% or more, and preferably 80% or more.

[0165] As a medium for induction culture, for example, a human ES / iPS medium such as TeSR2 (STEMCELL Technologies) can be used. However, the medium is not limited thereto, and various stem cell media can be used. For example, Primate ES Cell Medium, Reprostem, ReproFF, ReproFF2, ReproXF (Reprocell), mTeSR1, TeSRE8, ReproTeSR (STEMCELL Technologies), PluriSTEM (registered trademark) Human ES / iPS Medium (Merck), NutriStem (registered trademark) XF / FF Culture Medium for Human iPS and ES Cells, Pluriton reprogramming medium (Stemgent), PluriSTEM (registered trademark), Stemfit AK02N, Stemfit AK03 (Ajinomoto), ESC-Sure (registered trademark) serum and feeder free medium for hESC / iPS (Applied StemCell), L7 (registered trademark) hPSC Culture System (LONZA), Primate ES Cell Medium (ReproCELL), and Puel (I Peace, Inc.) may be used. The medium does not contain activin A, for example.

[0166] The gel medium is prepared, for example, by adding deacylated gellan gum to the above medium to a final concentration of 0.5% to 0.001% by weight, 0.1% to 0.005% by weight, or 0.05% to 0.01% by weight.

[0167] When maintaining and culturing stem cells in an undifferentiated state, the medium contains b-FGF at 10 ng / ml or more, preferably 40 ng / ml or more. Whether or not stem cells maintain an undifferentiated state can be confirmed by, for example, whether the positive rate of any of TRA1-60, OCT3 / 4, SSEA3, SSEA4, TRA1-81, LIN28, and NANOG is 30% or more, 50% or more, preferably 80% or more.

[0168] The stem cell extract may be a liquid. That is, the extract may be an extract solution. The stem cell extract may be a stem cell paste. The stem cell paste is obtained by grinding the stem cells. The stem cell extract may be a lyophilized product. The stem cell extract may also be a powder. Alternatively, the stem cell extract may be a stem cell lysate.

[0169] The stem cell culture medium or the supernatant of the stem cell culture medium contains secretions from the stem cells. Examples of secretions from the stem cells include extracellular vesicles, cytokines, chemokines, and growth factors. Extracellular vesicles are vesicles having a lipid bilayer structure. Extracellular vesicles are also called membrane vesicles. Examples of extracellular vesicles include exosomes and microvesicles. Exosomes are granular substances with a diameter of 30 nm to 150 nm. Exosomes are formed by inward budding of the late endosomal membrane, and then fuse with the cell membrane to form complete particles, which are secreted outside the cell by exocytosis. Exosomes contain lipids and proteins derived from the cell membrane on their surface. Exosomes also contain nucleic acids and proteins derived from the inside of the cell.

[0170] The secretion process of exosomes will be further explained. When endocytosis occurs in cells, early endosomes are formed, followed by late endosomes. At this time, vesicles formed by budding the endosomal membrane inward are called intraluminal membrane vesicles (ILVs). Endosomes in which intraluminal membrane vesicles are gathered are called multivesicular endosomes (MVBs). When multivesicular endosomes fuse with the cell membrane, the intraluminal membrane vesicles in the multivesicular endosomes are exocytized to the outside of the cell. The intraluminal membrane vesicles released to the outside of the cell are exosomes. Involvement of ceramide, which is important for vesicle budding into the endosomal membrane, Hrs, a subunit of the endosomal sorting complex for transport (ESCRT), and RAB family members such as RAB11 and RAB35, which are low molecular weight G proteins involved in the intracellular transport of vesicles, has been reported in this series of secretion processes. In addition, involvement of membrane-associated proteins, such as SNARE, has been reported when multivesicular endosomes fuse with the cell membrane to cause exocytosis.

[0171] The method for recovering exosomes from the culture medium is not particularly limited. For example, the culture medium may be centrifuged by ultracentrifugation or density gradient centrifugation to recover exosomes from the culture medium. In ultracentrifugation, the culture medium may be centrifuged at 10,000-100,000 g for 30 minutes or more to isolate exosomes from the culture medium. Exosomes may be recovered by size exclusion chromatography. Exosomes may be co-precipitated with polyethylene glycol (PEG) to recover exosomes. Exosomes may be isolated from serum or culture supernatant by immunoprecipitation using antibodies that specifically recognize CD9, CD63, and CD81.

[0172] Exosomes may be collected by affinity purification using a protein specifically expressed in exosomes. For example, the phospholipid phosphatidylserine is expressed on the membrane surface of exosomes. Tim4, an exosome receptor expressed in macrophages, binds to the phospholipid phosphatidylserine in a calcium-dependent manner. Therefore, it is possible to collect and isolate exosomes from the culture medium by preparing magnetic beads to which the extracellular domain of Tim4 is bound, immobilizing exosomes on the surface of the magnetic beads via the extracellular domain of Tim4, and then eluting the exosomes from the magnetic beads using a chelating agent.

[0173] The microvesicles have a diameter of 100 nm to 1000 nm and are generated by the outward budding and separation of the cell membrane. The secretion products of stem cells may be, for example, ectosomes, secretory microvesicles, oncosomes, and prostasomes. The secretion products of stem cells may be, for example, peptides, proteins, growth factors, cytokines, and hormones. The proteins have a molecular weight of, for example, 3 kDa or more, 10 kDa or more, or 20 kDa or more.

[0174] The secretion product of the stem cells may be contained in the supernatant of the culture solution of the stem cells. The pharmaceutical composition or cosmetic composition according to the embodiment may contain an isolated secretion product. The pharmaceutical composition or cosmetic composition according to the embodiment may contain a concentrated secretion product. Examples of the method for isolating and concentrating the secretion product include a freeze-drying method, an ultrafiltration method, a precipitation and resuspension method, and a gel filtration method. Examples of the method for precipitating proteins include an ammonium sulfate precipitation method, a trichloroacetic acid method, an acetone precipitation method, and an ammonium acetate-containing methanol precipitation method.

[0175] Examples of marker proteins for extracellular vesicles include the tetraspanin family (CD9, CD63, CD81), the integrin family (ITGA, ITGB), ESCRT complex binding proteins (TSG101, ALIX), lipid raft markers (Flotillin-1 / 2), and heat shock proteins (HSP70). Therefore, extracellular vesicles may be isolated using antibodies that specifically recognize these marker proteins for extracellular vesicles.

[0176] The secretory product may be collected from the supernatant of the culture medium of the stem cells.The secretory product may be collected from the supernatant of the culture medium during the maintenance culture of the stem cells.

[0177] The present inventors have found for the first time that stem cells, stem cell extracts, stem cell culture media, supernatants of culture fluids during stem cell induction, and supernatants of culture fluids of stem cells decompose fat. Here, fat decomposition includes fat burning and fat consumption. Therefore, stem cells, stem cell extracts, stem cell culture media, supernatants of culture fluids during stem cell induction, and supernatants of culture fluids of stem cells can be used as fat decomposition agents, anti-obesity drugs, obesity improvement drugs, fat decomposition promoters, and diet agents. It is also known that blood fat increases in diabetes. Therefore, stem cells, stem cell extracts, stem cell culture media, supernatants of culture fluids during stem cell induction, and supernatants of culture fluids of stem cells can be used as therapeutic or preventive drugs for diabetes. The fat is, for example, neutral fat. Diabetes may be type 1 diabetes, type 2 diabetes, diabetes due to other specific mechanisms, diseases, or gestational diabetes.

[0178] The present inventors have found for the first time that stem cells, stem cell extracts, stem cell culture media, supernatants of culture fluids during stem cell induction, and supernatants of culture fluids of stem cells promote differentiation induction from myoblasts to myotubes. Therefore, stem cells, stem cell extracts, stem cell culture media, supernatants of culture fluids during stem cell induction, and supernatants of culture fluids of stem cells can be used as agents for promoting differentiation induction from myoblasts to myotubes. In addition, myotubes form muscle fibers and become muscles. Therefore, stem cells, stem cell extracts, stem cell culture media, supernatants of culture fluids during stem cell induction, and supernatants of culture fluids of stem cells can be used as agents for promoting muscle formation, maintaining muscle mass, and increasing muscle mass.

[0179] The present inventors have found for the first time that stem cells, stem cell extracts, stem cell culture media, supernatants of culture media during stem cell induction, and supernatants of culture media during stem cell induction promote endothelial cells to produce prostacyclin. Therefore, stem cells, stem cell extracts, stem cell culture media, supernatants of culture media during stem cell induction, and supernatants of culture media during stem cell induction can be used as prostacyclin production promoters. Prostacyclin is also called prostaglandin I2 (PGI2). Prostacyclin activates adenylate cyclase and promotes the production of cyclic adenosine monophosphate (cAMP). Therefore, stem cells, stem cell extracts, stem cell culture media, supernatants of culture media during stem cell induction, and supernatants of culture media during stem cell induction can be used as adenylate cyclase activators and cyclic adenosine monophosphate (cAMP) production promoters.

[0180] In addition, prostacyclin has a platelet aggregation inhibitory effect and a vascular smooth muscle dilating effect. Therefore, the stem cells, stem cell extracts, stem cell culture medium, the supernatant of the culture medium during stem cell induction, and the supernatant of the culture medium of stem cells can be used as a platelet aggregation inhibitor, a vasodilator, a therapeutic or preventive drug for hypertension, and a therapeutic or preventive drug for arteriosclerosis. Furthermore, prostacyclin inhibits the proliferation of vascular endothelial cells. Therefore, the stem cells, stem cell extracts, stem cell culture medium, the supernatant of the culture medium during stem cell induction, and the supernatant of the culture medium of stem cells can be used as a vascular endothelial cell proliferation inhibitor.

[0181] The present inventor has found for the first time that stem cells, stem cell extracts, stem cell culture medium, supernatant of culture medium during stem cell induction, and supernatant of culture medium during stem cell induction promote chondrocytes to produce hyaluronic acid.Therefore, stem cells, stem cell extracts, stem cell culture medium, supernatant of culture medium during stem cell induction, and supernatant of culture medium during stem cell induction can be used as chondrocytes hyaluronic acid production promoters.In addition, hyaluronic acid protects cartilage and has lubricating, buffering, and anti-inflammatory effects on joints.Therefore, stem cells, stem cell extracts, stem cell culture medium, supernatant of culture medium during stem cell induction, and supernatant of culture medium during stem cell induction can be used as chondrocyte proliferation promoters, treatment or prevention drugs for cartilage tissue wounds, treatment or prevention drugs for joint pain, analgesics, treatment, improvement, or prevention drugs for joints, joint diseases, or arthritis, inflammation suppressants, and inflammation alleviators. Examples of arthropathy include osteoarthritis, osteoarthritis, temporomandibular joint disorder, femoral head necrosis, rheumatoid arthritis, chondritis, tennis elbow, golf elbow, baseball elbow, hernia, cartilage injury, and meniscus injury.Arthropathy is, for example, osteoarthritis.The hyaluronic acid production promoter of chondrocytes, the proliferation promoter of chondrocytes, the treatment or prevention drug for cartilage tissue injury, the treatment or prevention drug for joint pain, the analgesic, and the treatment, improvement, or prevention drug for joints, joint diseases, or arthritis may further contain hyaluronic acid.

[0182] The present inventor also found for the first time that stem cells, stem cell extracts, stem cell culture medium, supernatant of culture medium during stem cell induction, and supernatant of culture medium of stem cells activate chondrocytes, increase the survival rate of chondrocytes, promote proliferation, and rejuvenate chondrocytes.Therefore, stem cells, stem cell extracts, stem cell culture medium, supernatant of culture medium during stem cell induction, and supernatant of culture medium of stem cells can be used as a chondrocyte activator, a chondrocyte survival rate improver, a chondrocyte proliferation promoter, a chondrocyte protection agent, a chondrocyte rejuvenation agent, a cartilage regenerator, and a joint, joint disease, or arthritis treatment, improvement, or prevention drug.The chondrocyte activator, chondrocyte survival rate improver, chondrocyte proliferation promoter, chondrocyte rejuvenation agent, cartilage regenerator, and joint, joint disease, or arthritis treatment, improvement, or prevention drug may further contain hyaluronic acid.

[0183] In addition, the anti-obesity effect, muscle mass increasing effect, vasodilatory effect, cartilage protection effect, etc. suppress aging and lead to the treatment and prevention of lifestyle-related diseases and adult diseases. Therefore, stem cells, stem cell extracts, stem cell culture media, culture supernatants during stem cell induction, and stem cell culture supernatants are anti-aging agents, therapeutic or preventive agents for lifestyle-related diseases, and / or therapeutic or preventive agents for adult diseases.

[0184] The present inventor also found for the first time that stem cells, stem cell extracts, stem cell culture media, supernatants of culture fluids during stem cell induction, and supernatants of culture fluids of stem cells amplify the expression of GDF11, MANF, TIMP2, SPARCL1, THBS4, NAMPT, NANOG, and TERT, and suppress the expression of p53 and p16. GDF11, MANF, TIMP2, SPARCL1, THBS4, NAMPT, NANOG, p53, and p16 are expressed, for example, in cells of the entire individual, but are not limited thereto. Examples of cells include skin cells. Examples of skin cells include fibroblasts, cells of the epidermis layer, and cells of the dermis layer. It is known that the expression of GDF11, MANF, TIMP2, SPARCL1, THBS4, NAMPT, and NANOG is high in humans and non-human animals when they are young, and decreases as they age. GDF11, MANF, TIMP2, SPARCL1, THBS4, NAMPT, and NANOG are also called rejuvenation factors or rejuvenation-related factors. Therefore, by increasing the expression of at least one of GDF11, MANF, TIMP2, SPARCL1, THBS4, NAMPT, and NANOG, it is possible to rejuvenate humans and non-human animals and extend the life span of humans and non-human animals. TERT promotes telomere elongation and inhibits telomere shortening. Therefore, stem cells, stem cell extracts, stem cell culture media, supernatants of culture fluids during stem cell induction, and supernatants of stem cell culture fluids can be used as anti-aging agents, rejuvenation agents, expression amplifiers of rejuvenation factors, expression amplifiers of rejuvenation-related factors, expression amplifiers of at least one of GDF11, MANF, TIMP2, SPARCL1, THBS4, NAMPT, and NANOG, expression amplifiers of TERT, telomere elongation agents, and telomere shortening inhibitors. In addition, p53 and p16 are known to be cellular senescence markers, and therefore, by increasing the expression of at least one of p53 and p16, it is possible to suppress cellular senescence.Therefore, stem cells, stem cell extracts, stem cell culture media, supernatants of culture media during stem cell induction, and supernatants of stem cell culture media can be used as anti-aging agents, rejuvenating agents, inhibitors of the expression of aging-related factors, inhibitors of the expression of cellular aging-related factors, inhibitors of the expression of individual aging-related factors, inhibitors of the expression of aging markers, inhibitors of the expression of cellular aging markers, inhibitors of p53 expression, inhibitors of p16 expression, inhibitors of the p53 pathway, inhibitors of the p16 pathway, inhibitors of the p19-p52 pathway, and inhibitors of the p16-Rb pathway.

[0185] The pharmaceutical composition and cosmetic composition according to the embodiment include an effective amount of stem cells, stem cell extracts, stem cell culture medium, supernatant of culture fluid during stem cell induction, supernatant of culture fluid of stem cells, or secretion of stem cells. Here, the effective amount refers to an amount that can exert efficacy as a pharmaceutical composition or cosmetic composition. The effective amount is appropriately set according to the age of the patient, the target disease, the presence or absence of other effective ingredients, and the amount of other compounds. The pharmaceutical composition and cosmetic composition according to the embodiment may be repeatedly administered to the affected area and the target site. Examples of the administration route of the pharmaceutical composition and cosmetic composition according to the embodiment include oral administration, subcutaneous administration by injection, intramuscular administration by injection, intravenous administration by injection, intrathecal administration by injection, sublingual administration, oral mucosal administration, rectal administration, vaginal administration, administration to the eye, administration to the ear, nasal administration, inhalation administration, spray administration, and transdermal administration.

[0186] The pharmaceutical composition and / or cosmetic composition according to the embodiment may include a pharma- ceutically acceptable carrier, an excipient, a disintegrant, a buffer, an emulsifier, a suspending agent, a soothing agent, a stabilizer, a preservative, a preservative, and physiological saline. Examples of excipients include lactose, starch, sorbitol, D-mannitol, and sucrose. Examples of disintegrants include carboxymethylcellulose and calcium carbonate. Examples of buffers include phosphates, citrates, and acetates. Examples of emulsifiers include gum arabic, sodium alginate, and tragacanth. The pharmaceutical composition according to the embodiment may be included in an injection solution and an infusion solution. The pharmaceutical composition according to the embodiment may be injected into an affected area. The pharmaceutical composition according to the embodiment may be a research reagent.

[0187] Examples of suspending agents include glyceryl monostearate, aluminum monostearate, methylcellulose, carboxymethylcellulose, hydroxymethylcellulose, and sodium lauryl sulfate. Examples of soothing agents include benzyl alcohol, chlorobutanol, and sorbitol. Examples of stabilizers include propylene glycol and ascorbic acid. Examples of preservatives include phenol, benzalkonium chloride, benzyl alcohol, chlorobutanol, and methylparaben. Examples of preservatives include benzalkonium chloride, paraoxybenzoic acid, and chlorobutanol.

[0188] In addition, the pharmaceutical composition and / or cosmetic composition according to the embodiment may contain water, alcohol, surfactants (cationic, anionic, nonionic, amphoteric surfactants, etc.), moisturizers (glycerin, 1,3-butylene glycol, propylene glycol, propanediol, pentanediol, polyquaternium, amino acids, urea, pyrrolidone carboxylates, nucleic acids, monosaccharides, oligosaccharides, etc., and derivatives thereof, etc.), thickeners (polysaccharides, polyacrylates, carboxyvinyl polymers, polyvinylpyrrolidone, polyvinyl alcohol, chitin, chitosan, alginate, etc.), and the like. Acids, carrageenan, xanthan gum, methylcellulose, etc., and their derivatives, wax, petrolatum, hydrocarbon saturated fatty acids, unsaturated fatty acids, silicone oils, etc., and their derivatives, triglycerides such as tri(caprylic / capric acid)glyceryl and trioctanoic acid glyceryl, ester oils such as isopropyl stearate, natural oils and fats (olive oil, camellia oil, avocado oil, almond oil, cacao butter, evening primrose oil, grape seed oil, macadamia nut oil, eucalyptus oil, rosehip oil, squalane, orange roughy oil, lanolin , and ceramides, etc.), preservatives (oxybenzoic acid derivatives, dehydroacetate, photosensitizers, sorbic acid, phenoxyethanol, etc., and their derivatives, etc.), disinfectants (sulfur, triclocarbanilide, salicylic acid, zinc pyrithione, hinokitiol, etc., and their derivatives, etc.), ultraviolet absorbers (para-aminobenzoic acid, methoxycinnamic acid, etc., and their derivatives, etc.), anti-inflammatory agents (allantoin, bisabolol, ε-aminocaproic acid, acetylphanesylcysteine, glycyrrhizic acid, etc., and their derivatives, etc.), antioxidants (Tocopherol, BHA, BHT, astaxanthin, etc. and their derivatives, etc.), chelating agents (EDTA, hydroxyethanediphosphonic acid, etc. and their derivatives, etc.), animal and plant extracts (Angelica angelica, aloe, star anise, Scutellaria baicalensis, Phellodendron bark, seaweed, Chinese quince, chamomile, licorice, kiwi, cucumber, mulberry, birch, angelica tree, garlic, peony, hops, horse chestnut, lavender, rosemary, eucalyptus, milk, various peptides, placenta, royal jelly, Euglena extract, hydrolyzed Euglena extract, and Euglena oil, etc.),and refined or fermented products of these ingredients), pH adjusters (inorganic acids, inorganic acid salts, organic acids, organic acid salts, etc., and their derivatives, etc.), vitamins (vitamin A, vitamin B, vitamin C, vitamin D, ubiquinone, nicotinamide, etc., and their derivatives, etc.), fermentation liquid of yeast, koji mold, and lactic acid bacteria, Galactomyces culture liquid, skin whitening agents (tranexamic acid, cetyl tranexamate hydrochloride, 4-n-butylresorcinol, arbutin, kojic acid, ellagic acid, licorice flavonoids, niacinamide , and vitamin C derivatives, etc.), ceramides and ceramide derivatives, anti-wrinkle agents (retinol, retinal, and derivatives thereof, nicotinamide, oligopeptides, etc., and derivatives thereof, natural and synthetic ingredients having neutrophil elastase inhibitory, and MMP-1 and MMP-2 inhibitory effects, etc.), titanium oxide, talc, mica, silica, zinc oxide, iron oxide, silicon, and powders processed from these, etc., can be blended within a range that achieves the objectives of the pharmaceutical composition and cosmetic composition according to the embodiment.

[0189] In addition, the components that can be added to the pharmaceutical composition and / or cosmetic composition according to the embodiment are not limited to the above, and can be freely selected as long as they are components that can be used in pharmaceutical compositions and / or cosmetic compositions. When the pharmaceutical composition and / or cosmetic composition according to the embodiment is used as a poultice, in addition to the above components, a base (kaolin, bentonite, etc.) and a gelling agent (polyacrylate, polyvinyl alcohol, etc.) can be blended within a range that achieves the purpose. When the pharmaceutical composition and / or cosmetic composition according to the embodiment is used as a bath additive, a sulfate, a bicarbonate, a borate, a colorant, and a moisturizing agent can be appropriately blended within a range that achieves the purpose, and the composition may be prepared into a powder type or a liquid type. The pharmaceutical composition and / or cosmetic composition according to the embodiment may be a skin application composition. EXAMPLES

[0190] Example 1: Preparation of test substance Test substances 1 to 7 shown in Table 1 were prepared. Test substance 1 contained only Dulbecco's modified Eagle's medium (DMEM). Test substance 2 contained only stem cell medium (Puel, I Peace, Inc.). [Table 1]

[0191] Test substance 3 was prepared by the following method. A serum-free and animal-derived component-free hematopoietic cell culture medium (Stemspan ACF, STEMCELL TECHNOLOGIES) was supplemented with recombinant protein FLT3 ligand (PEPROTECH), recombinant human TPO (PEPROTECH), recombinant human IL-6 (PEPROTECH), recombinant human G-CSF (PEPROTECH), and recombinant human SCF (PEPROTECH) to prepare a hematopoietic cell culture medium. 2 × 10 5 Human blood cells (peripheral blood mononuclear cells) were seeded on the wells of the 12-well dish. The wells were then placed in a CO2 incubator at 37°C to culture the cells in suspension.

[0192] Three days after the start of cell culture, hematopoietic cell medium was added to each well as needed. Six days after the start of cell culture, Oct3 / 4, SOX2, KLF4, and c-Myc were transfected into the cells in each well, after which hematopoietic cell medium was added to each well and the cells were cultured on a laminin 511-coated dish, which is a stem cell culture substrate.

[0193] Two days after infection, half of the culture medium in each well was replaced with stem cell medium every two days. 14 days after infection, it was confirmed that 10 or more iPS cell clusters had been induced, and the culture supernatant was collected. The collected culture supernatant was centrifuged at 400g to remove the cells, and the culture supernatant filtered through a filter was used as test substance 3.

[0194] Test substance 4 was prepared by the following method. Human iPS cells were cultured in a stem cell medium (Puel, I Peace, Inc.) on a petri dish for adhesion culture coated with laminin 511. The human iPS cells were passaged every week. During passage, the human iPS cells were treated with an ES cell dissociation solution (TrypLE Select, registered trademark, Thermo Fisher Scientific).

[0195] The human iPS cells maintained as described above were detached from the petri dish for adhesion culture using ES cell dissociation solution (TrypLE Select, registered trademark, Thermo Fisher Scientific). The detached human iPS cells were seeded on a dish coated with laminin (Nippi). The human iPS cells were then cultured for one week using stem cell medium (Puel, I Peace, Inc.) supplemented with 10 μmol / L of ROCK inhibitor. The medium was changed every day.

[0196] After confirming that the cells were 40% or more confluent, the medium was replaced with fresh culture medium, and the supernatant of the stem cell medium was collected two days later. The collected supernatant of the stem cell medium was centrifuged at 400g for 30 minutes, and the supernatant of the stem cell medium after centrifugation was filtered through a 0.22 μm filter. The filtered supernatant of the stem cell medium was used as test substance 4. In addition, it was confirmed that the iPS cells maintained in culture were positive for the undifferentiated markers NANOG, OCT3 / 4, and TRA 1-60.

[0197] Test substance 5 was prepared by the following method. iPS cells derived from human blood cells were established by two-dimensional culture. The iPS cells were passaged five times or more, and it was confirmed that no reprogramming factors remained in the iPS cells and that the iPS cells did not contain viruses. The iPS cells were then cryopreserved using a freezing solution (STEM-CELLBANKER).

[0198] A 30 mL bioreactor was prepared. 20 mL of stem cell medium (Puel, I Peace, Inc.) was placed in the bioreactor. 4 × 10 cells frozen in freezing solution (STEM-CELLBANKER) were placed in the bioreactor. 6 Undifferentiated iPS cells including single cells were seeded in a stem cell medium in a bioreactor, and three-dimensional stirring culture of iPS cells was started. After the cells formed clumps, the medium was stirred at a speed at which 20% to 60% of the clumps sank.

[0199] After 2 days, 10 mL of stem cell medium was added to the bioreactor. Stem cell medium was then added to the bioreactor every 2 days until cell clusters were formed. Seven days after seeding the iPS cells into the bioreactor, the cells were analyzed by flow cytometry and were positive for TRA1-60, a marker of pluripotent stem cells.

[0200] Next, the stem cell medium in which the iPS cells were suspended was filtered through a 0.22 μm mesh filter to remove cell aggregates. The filtered stem cell medium was then centrifuged at 1500 rpm for 5 minutes to precipitate the cells, and the supernatant of the stem cell medium after centrifugation was collected again and centrifuged at 3000 rpm for 3 minutes. The supernatant of the culture medium after centrifugation was filtered through a 0.22 μm filter. The supernatant of the culture medium after filtration was designated as test substance 5.

[0201] Test substance 6 was prepared by the following method. Human iPS cells used when Test substance 4 was prepared were peeled off from the dish using Triple Select, and the cells were centrifuged to obtain a pellet, which was then ground with a pestle (Pestle in G-Tube, Thermo Fisher Scientific). The paste of the ground human iPS cells was dissolved in DMEM / F12 to obtain a human iPS cell extract. The obtained human iPS cell extract was stored at 4°C for 1 to 24 hours, and then the human iPS cell extract was filtered using a filter. The filtered human iPS cell extract was frozen and stored. Before being used in the following examples, the human iPS cell extract was thawed and used as Test substance 6.

[0202] Test substance 7 was prepared by the following method. An extract of porcine iPS cells was prepared from porcine iPS cells induced under the same conditions as those for preparing test substance 4, except that pig-derived blood cells were used, by the same method as those for preparing test substance 6, and used as test substance 7.

[0203] In addition, isoproterenol (Cayman), known to have a lipolytic effect, was prepared as a positive control substance in Example 2. Insulin (Thermo Fisher Scientific), known to induce differentiation from myoblasts to myotubes, was prepared as a positive control substance in Example 3. Arachidonic acid (Sigma-Aldrich, A3411), known to promote the production of prostacyclin, was prepared as a positive control substance in Example 4. N-acetylglucosamine (Sigma-Aldrich, A8625), known to promote the production of hyaluronic acid, was prepared as a positive control substance in Example 5.

[0204] Example 2: Lipolysis Test Normal human subcutaneous preadipocytes (Lonza, PT-5020) were induced to sleep in a T-75 flask using growth medium (PGM-2, 10% FBS, 2 mM L-glutamine, 50 μg / mL gentamicin, 37 ng / mL amphotericin-B-added PBM, Lonza, PT-8002), and cultured in a CO2 incubator (37°C, 5% CO2 humidified, same below) until the cells were 60 to 80% confluent. Next, the normal human subcutaneous preadipocytes were detached from the T-75 flask using 0.25% trypsin-EDTA, and the normal human subcutaneous preadipocytes were centrifuged. Then, the normal human subcutaneous preadipocytes were seeded in a 96-well plate at 10,000 cells / 100 μL / well using growth medium, and cultured in a CO2 incubator.

[0205] The next day, the medium was replaced with 100 μL of differentiation medium (PGM-2, Lonza, PT-8002 supplemented with insulin, dexamethasone, indomethacin, and isobutyl-methylxanthine), and the cells were cultured in a CO2 incubator for 7 days to induce differentiation of normal human subcutaneous preadipocytes into adipocytes. Next, the medium was replaced with 100 μL of differentiation medium supplemented with test substances 1 to 7 at final concentrations of 5% and 20% and positive control substance at final concentration of 10 μmol / L, or with 100 μL of differentiation medium without test substance or positive control substance, and the cells were cultured in a CO2 incubator for 24 hours to treat the adipocytes with either test substances 1 to 7 or positive control substance. Photographs of the cells after treatment are shown in Figures 1, 2, and 3.

[0206] The culture supernatant was then collected, and the amount of free glycerol in the supernatant was measured using a lipolysis analysis kit (Cayman, 10009381). The amount of free glycerol corresponds to the amount of lipolysis. The measurement method followed the protocol attached to the kit. Specifically, the absorbance of the supernatant was measured at a wavelength of 540 nm using a multispectro microplate reader (Varioskan Flash, Thermo Fisher Scientific), and the relative free glycerol concentration was calculated. The control group was a group to which the test substance was not added, and the control group was compared with the group to which the test substance and positive control substance were added using a two-tailed Dunnett's test, and a significant difference was determined when P<0.05.

[0207] When the final concentration of test substances 1 to 7 was 5%, as shown in Figures 4 and 5, the groups to which test substances 2 to 7 were added had a significantly higher amount of lipolysis compared to the control. The group to which test substance 1, which is DMEM with a final concentration of 5%, was added did not have a significantly higher amount of lipolysis compared to the control. When the final concentration of test substances 1 to 6 was 20%, as shown in Figures 6 and 7, the groups to which test substances 2 to 6 were added had a significantly higher amount of lipolysis compared to the control. The group to which test substance 1, which is DMEM with a final concentration of 20%, was added did not have a significantly higher amount of lipolysis compared to the control. As shown in Figures 8 and 9, the group to which isoproterenol, which is known to have a lipolytic effect, was added had a significantly higher amount of lipolysis compared to the control, indicating that the experimental system was correct. These results indicate that the stem cell medium, the medium used for stem cell induction culture, the medium used for stem cell maintenance culture, and the stem cell extract decompose fat.

[0208] (Example 3: Myotube differentiation promotion test) Normal human skeletal muscle myoblasts (Lonza, CC-2580) were put to sleep in a T-75 flask using growth medium (SkGM-2, Lonza, CC-3245), and cultured in a CO2 incubator until the normal human skeletal muscle myoblasts were about 60% confluent. Next, the normal human skeletal muscle myoblasts were detached from the T-75 flask using 0.05% trypsin-EDTA and phenol red. Then, the normal human skeletal muscle myoblasts were seeded in a 96-well plate at 7,500 cells / 100 μL / well using growth medium, and cultured in a CO2 incubator.

[0209] The next day, the medium was replaced with 100 μL of differentiation medium (DMEM:F12 supplemented with 2% horse serum) containing test substances 1 to 7 at final concentrations of 5% and 20% and the positive control substance at a final concentration of 50 nmol / L, or with 100 μL of differentiation medium without test substances or positive control substances, and the cells were cultured in a CO2 incubator for 3 days to induce differentiation of normal human skeletal muscle myoblasts into myotubes.

[0210] Thereafter, the medium was removed from the wells, and 100 μL of 4% paraformaldehyde was added to the wells at 4° C. and incubated for 15 minutes at 4° C. to fix the cells. Next, the wells were washed three times with 100 μL of Dulbecco's PBS, and 100 μL of blocking / membrane permeabilization solution (Dulbecco's PBS supplemented with 3% bovine serum albumin and 0.3% Triton-X 100) was added to the wells and incubated at room temperature for 30 minutes.

[0211] Next, the solution in the wells was replaced with a solution containing anti-myosin heavy chain (MHC) antibody (Affymetrix) and incubated overnight at 4°C. Myosin heavy chain is a differentiation marker of myotube cells. After washing the wells three times with buffer, secondary antibody (goat anti-mouse IgG2b antibody, Alexa Fluor 555, Thermo Fisher Scientific) and nuclear stain (Hoechst 33342, Thermo Fisher Scientific) were added to the wells and incubated at room temperature for 2 hours. After washing the wells three times with buffer, buffer was added to the wells and the central nine fields of each well were photographed using a confocal imaging system (Operetta CLS, Perkin Elmer) with a 10x objective lens.

[0212] The photographed images are shown in Figures 10, 11, and 12. From the photographed images, the number of cell nuclei and the number of myosin heavy chain-positive nuclei were counted, and the differentiation index I (%) given by the following formula (1) was calculated. A higher differentiation index I indicates that myotube differentiation was promoted. I=N M / N T ×100 (1) In equation (1), N M is the number of myosin heavy chain-positive nuclei, N T represents the total number of nuclei.

[0213] When the final concentration of test substances 1 to 7 was 5%, as shown in Figures 13 and 14, the groups to which test substances 3 to 7 were added had a significantly higher differentiation index than the group to which test substance 2 was added, which was only a stem cell medium. When the final concentration of test substances 1 to 7 was 20%, as shown in Figures 15 and 16, the groups to which test substances 3 to 7 were added had a significantly higher differentiation index than the group to which test substance 2 was added, which was only a stem cell medium. As shown in Figures 17 and 18, the group to which insulin, which is known to induce differentiation from myoblasts to myotubes, was added had a significantly higher differentiation index than the control, indicating that the experimental system was correct. These results indicate that the medium used for stem cell induction culture, the medium used for stem cell maintenance culture, and the stem cell extract promote the induction of differentiation from normal human skeletal muscle myoblasts to myotubes.

[0214] Example 4: Prostacyclin production test Normal human umbilical vein endothelial cells (Lonza, CC2519AS) were induced to sleep in a T-75 flask using medium (EGM-2, Lonza, CC-3162), and cultured in a CO2 incubator until the cells were 60 to 80% confluent. Next, the normal human umbilical vein endothelial cells were detached from the T-75 flask using 0.05% trypsin-EDTA and phenol red. Then, the normal human umbilical vein endothelial cells were seeded in a 96-well plate at 10,000 cells / 100 μL / well using medium, and cultured in a CO2 incubator.

[0215] The next day, the medium was replaced with 100 μL of medium containing test substances 1 to 7 at final concentrations of 5% and 20% and the positive control substance at a final concentration of 10 μmol / L, or 100 μL of medium without test substances or positive control substance, and the cells were cultured in a CO2 incubator for 24 hours.

[0216] The culture supernatant was collected, and the amount of 6-ketoprostaglandin F1α, a metabolic product of prostacyclin, was measured using an ELISA kit (Abcam, ab133023) and a multispectro microplate reader (Varioskan Flash, Thermo Fisher Scientific).

[0217] When the final concentration of test substances 1 to 7 was 5%, as shown in Figures 19 and 20, the groups to which test substances 3 to 7 were added had significantly higher levels of 6-ketoprostaglandin F1α than the control group to which no test substances were added. When the final concentration of test substances 1 to 7 was 20%, as shown in Figures 21 and 22, the groups to which test substances 3 to 7 were added had significantly higher levels of 6-ketoprostaglandin F1α than the control group to which no test substances were added. As shown in Figures 23 and 24, the groups to which arachidonic acid, which is known to promote the production of prostacyclin, were added had significantly higher levels of 6-ketoprostaglandin F1α than the control group, indicating that the experimental system was correct. These results indicate that the medium used for the induction culture of stem cells, the medium used for the maintenance culture of stem cells, and the stem cell extract promote the production of prostacyclin by normal human umbilical vein endothelial cells.

[0218] (Example 5: Hyaluronic acid production test) Human chondrocyte cell line (SW1353, ATCC, HTB-94) was put to sleep in a T-75 flask using growth medium (DMEM / F-12 supplemented with 10% FBS and 1% penicillin-streptomycin mixed solution) and cultured in a CO2 incubator until the cells were 60 to 80% confluent. Next, the human chondrocytes were detached from the T-75 flask using 0.25% trypsin-EDTA and phenol red. Then, the human chondrocytes were seeded in a 96-well plate at 10,000 cells / 100 μL / well using growth medium and cultured in a CO2 incubator.

[0219] The next day, the medium was replaced with 100 μL of assay medium (DMEM / F-12 supplemented with 1% FBS, 1% penicillin-streptomycin mixed solution) supplemented with test substances 1 to 7 at final concentrations of 5% and 20% and positive control substance at a final concentration of 10 mmol / L, or with 100 μL of assay medium without test substances or positive control substance, and the human chondrocytes were cultured in a CO2 incubator for 24 hours or 72 hours.

[0220] The culture supernatant was collected, and the amount of hyaluronic acid was measured using an ELISA kit (Hyaluronan Quantikine ELISA Kit, R&D Systems) and a multispectro microplate reader (Varioskan Flash, Thermo Fisher Scientific).

[0221] When human chondrocytes were cultured for 24 hours and the final concentration of test substances 1 and 3 to 7 was 5%, the group to which test substances 3 to 6 were added had a significantly higher amount of hyaluronic acid produced compared to the control to which no test substance was added, as shown in Figures 25 and 26. When human chondrocytes were cultured for 24 hours and the final concentration of test substances 1 and 3 to 7 was 20%, the group to which test substances 3 to 6 were added had a significantly higher amount of hyaluronic acid produced compared to the control to which no test substance was added, as shown in Figures 27 and 28. As shown in Figures 29 and 30, the group to which N-acetylglucosamine, which is known to promote the production of hyaluronic acid, was added had a significantly higher amount of hyaluronic acid produced compared to the control, indicating that the experimental system was correct. These results indicate that the medium used for induction culture of stem cells, the medium used for maintenance culture of stem cells, and the stem cell extract promote the production of hyaluronic acid by human chondrocytes.

[0222] When human chondrocytes were cultured for 72 hours and the final concentration of test substances 1 and 3 to 7 was 5%, the group to which test substances 3 to 5 were added had a significantly higher amount of hyaluronic acid produced compared to the control to which no test substance was added, as shown in Figures 31 and 32. When human chondrocytes were cultured for 72 hours and the final concentration of test substances 1 and 3 to 7 was 20%, the group to which test substances 3 to 7 were added had a significantly higher amount of hyaluronic acid produced compared to the control to which no test substance was added, as shown in Figures 33 and 34. As shown in Figures 35 and 36, the group to which N-acetylglucosamine, which is known to promote the production of hyaluronic acid, was added had a significantly higher amount of hyaluronic acid produced compared to the control, indicating that the experimental system was correct. These results indicate that the medium used for induction culture of stem cells, the medium used for maintenance culture of stem cells, and the stem cell extract promote the production of hyaluronic acid by human chondrocytes.

[0223] When human chondrocytes were cultured for 24 hours and the final concentration of test substance 2 was 5%, the group to which test substance 2 was added had a significantly higher amount of hyaluronic acid produced compared to the control to which no test substance was added, as shown in Figures 37 and 38. When human chondrocytes were cultured for 24 hours and the final concentration of test substance 2 was 20%, the group to which test substance 2 was added had a significantly higher amount of hyaluronic acid produced compared to the control to which no test substance was added, as shown in Figures 39 and 40. These results indicate that the stem cell medium promotes the production of hyaluronic acid by human chondrocytes.

[0224] When human chondrocytes were cultured for 72 hours and the final concentration of test substance 2 was 5%, the group to which test substance 2 was added had a significantly higher amount of hyaluronic acid produced compared to the control to which no test substance was added, as shown in Figures 41 and 42. When human chondrocytes were cultured for 72 hours and the final concentration of test substance 2 was 20%, the group to which test substance 2 was added had a significantly higher amount of hyaluronic acid produced compared to the control to which no test substance was added, as shown in Figures 43 and 44.

[0225] In addition, the cell viability was confirmed by the WST-8 method using the well plate after collecting the culture supernatant. 100 μL of differentiation medium containing 10% viable cell measurement reagent SF was added to the well, and the cells were incubated in a CO2 incubator. The increase rate of absorbance at a wavelength of 450 nm per 60 minutes was measured using a multispectro microplate reader (Varioskan Flash, Thermo Fisher Scientific), and the relative absorbance was taken as the relative cell viability. When human chondrocytes were cultured for 24 hours, as shown in Figures 45 to 50, the survival rate of human chondrocytes added with test substances 2 to 7 was higher than that of test substance 1, and it was shown that the medium used for induction culture of stem cells, the medium used for maintenance culture of stem cells, and the stem cell extract increased the survival rate and promoted the proliferation of human chondrocytes. Even when human chondrocytes were cultured for 72 hours, as shown in Figures 51 to 56, the survival rate of human chondrocytes added with test substances 2 to 7 was higher than that of test substance 1, demonstrating that the medium used for stem cell induction culture, the medium used for stem cell maintenance culture, and the stem cell extract activate human chondrocytes, increase the survival rate of human chondrocytes, promote their proliferation, and rejuvenate human chondrocytes.

[0226] (Reference example) Human iPS cells were cultured according to the example described in JP 2016-128396 A. That is, human iPS cells were cultured on feeder cells on a petri dish for adhesion culture using the same stem cell medium as in Example 1 to maintain adhesion. Human iPS cells were passaged every week. During passage, human iPS cells were treated with a detachment solution containing 0.25% trypsin, 0.1 mg / mL collagenase IV, 1 mmol / L CaCl2, and 20% KSR.

[0227] The human iPS cells cultured as described above were detached from the petri dish for adhesion culture using an ES cell dissociation solution (TrypLE Select, registered trademark, Thermo Fisher Scientific). The detached human iPS cells were cultured in suspension for one week in non-gelled human iPS cells placed in a petri dish for non-adherent culture. As a result, embryoid bodies (EBs) were formed. The formed embryoid bodies were seeded on a petri dish for adhesion culture and outgrown for one week in DMEM containing 10% FBS and 1% Anti-Anti (registered trademark, antimycotic agent).

[0228] Next, the cells were detached from the petri dish for adhesion culture using 0.05% trypsin-EDTA solution, and the cells divided into single cells were seeded on a new petri dish for adhesion culture. After that, the cells were cultured for one week using DMEM containing 10% FBS as the medium.

[0229] After confirming that the cells were 70% to 80% or more confluent, the cells were observed. A photograph of the cells cultured in this reference example is shown in FIG. 57(a). Usually, the morphology of undifferentiated iPS cells is as shown in the photograph in FIG. 57(b). Therefore, it was morphologically observed that the cells cultured in this reference example did not maintain an undifferentiated state. In addition, after culturing the cells according to this reference example for 21 days, the cells were treated with an anti-OCT3 / 4 antibody labeled with a fluorescent reagent and an anti-NANOG antibody labeled with a fluorescent reagent, and the result of observing the cells under a microscope is shown in FIG. 58. FIG. 58(a) shows a photograph of the cells observed without using excitation light. FIG. 58(b) shows a photograph of the cells observed with excitation light corresponding to the fluorescent reagent bound to the anti-OCT3 / 4 antibody. FIG. 58(c) shows a photograph of the cells observed with excitation light corresponding to the fluorescent reagent bound to the anti-NANOG antibody. No fluorescence was observed in FIG. 58(b) and FIG. 58(c), and it was confirmed that the cells were OCT3 / 4 negative and NANOG negative. Furthermore, when the cells were examined using a flow cytometer, it was confirmed that the cultured cells were negative for the undifferentiated markers NANOG, OCT3 / 4, and TRA 1-60, confirming that the cells did not maintain an undifferentiated state but were differentiated.

[0230] (Example 6: Treatment test of rat model of osteoarthritis) Five-week-old male rats were prepared. Each rat was anesthetized by isoflurane inhalation, and 50 μL of 20 mg / mL monoiodoacetate (MIA) was administered into the left joint cavity using a 30G needle and a microsyringe to prepare a rat model of left hindlimb arthropathy. Three days after administration of MIA, the rats were divided into a test group and a control group, each consisting of six rats. The rats were divided so that there was no difference in the average left hindlimb load ratio between the test group and the control group.

[0231] The left hindlimb load ratio was measured by the following method. Each rat was placed in an incapacitance test device (Linton Instrumentation), and the left hindlimb load and the right hindlimb load were measured three times consecutively while the rat was in a resting state and in a stable position. The left hindlimb load ratio R (%) was then calculated using the following formula. R = L ÷ (L + R) × 100 In the above formula, L represents the average load of the left hind leg, and R represents the average load of the right hind leg.

[0232] On the 3rd, 7th, 11th, 15th, and 19th days after administration of MIA, each rat in the test group was anesthetized by isoflurane inhalation, and 50 μL of test substance 4 was administered into the left joint cavity using a 30G injection needle and a microsyringe. Also, on the 3rd day after administration of MIA, each rat in the control group was anesthetized by isoflurane inhalation, and 50 μL of saline was administered into the left joint cavity using a 30G injection needle and a microsyringe.

[0233] On the fifth day after administration of MIA, the left hind leg load ratio of the rats in the test group and the left hind leg load ratio of the rats in the control group were measured by the same method as above. As a result, as shown in FIG. 59, the left hind leg load ratio was increased in the rats in the test group administered with test substance 4 compared to the rats in the control group. This result indicates that the administration of test substance 4 has treated the left hind leg arthropathy and alleviated the pain in the left hind leg joint. In addition, when the rat joints were dissected, the number of chondrocytes was large in the joints administered with test substance 4, and the chondrocyte tissue remained predominantly in large amounts, indicating the improvement effect of the administered test substance 4 on the joint disease. This result indicates that test substance 4 improved the survival rate and proliferation rate of chondrocytes, and further improved the production amount of hyaluronic acid by chondrocytes.

[0234] (Example 7: Preparation of culture supernatant used in maintenance culture of iPS cells) 5×10 5 Human iPS cells from 100 cells were seeded on a laminin-coated 10 cm dish, and the iPS cells were cultured for 5 days using Puel medium until they became 80% confluent. The medium was replaced once every two days. On the fifth day after the cells were seeded, the medium was replaced with 10 mL of fresh Puel medium, and on the seventh day, the culture supernatant was collected in a 15 mL tube. The 15 mL tube containing the culture supernatant was centrifuged at 200 g for 5 minutes, and the supernatant was filtered using a 0.22 μm syringe filter to obtain the supernatant of the culture solution according to Example 7.

[0235] (Example 8: Expression of rejuvenation genes in vitro) A 6-well plate was prepared. 10 5 Human skin fibroblast cells were seeded and the skin cells were cultured in a skin cell medium. The next day, the skin cell medium was completely removed from the two wells, and 2 mL of the culture medium according to Example 7 was added to the first well. As a control, a skin cell medium was added to the second well. After two days, the skin cells were peeled off from the wells using trypsin, and the cell-containing liquid was collected in a 1.5 mL tube and centrifuged, after which the supernatant was removed.

[0236] RNA was extracted from skin cells using a total RNA purification kit (NucleoSpin, registered trademark, RNA Plus, MACHEREY-NAGEL). cDNA was synthesized from RNA using a cDNA synthesis reagent for real-time PCR (ReverTra Ace, registered trademark, qPCR RT Master Mix, TOYOBO), and qPCR was performed using a master mix for real-time PCR (THUNDERBIRD Next SYBR, registered trademark, qPCR Mix, TOYOBO). This allowed the RNA expression levels of GDF11, MANF, TIMP2, SPARCL1, THBS4, GAPDH, NAMPT2, NANOG, TERT, p53, and p16 in skin cells to be analyzed. The expression levels of each RNA were corrected using the expression level of GAPDH.

[0237] As a result, as shown in Figures 60, 61, 62, 63, and 64, it was shown that the expression levels of GDF11, MANF, TIMP2, SPARCL1, and THBS4 increased by adding the supernatant of the culture solution according to Example 7 to skin cells. It is known that these genes decrease throughout an individual as the individual ages. The increase in the expression levels of GDF11, MANF, TIMP2, SPARCL1, and THBS4 indicates that the addition of the supernatant of the culture solution to skin cells rejuvenates the skin cells. It also indicates that the supernatant of the culture solution has an anti-individual aging effect.

[0238] In addition, as shown in FIG. 65 and FIG. 66, it was shown that the expression levels of NAMPT2 and NANOG increased by adding the supernatant of the culture solution according to Example 7 to skin cells. It is known that these genes decrease throughout the entire individual as the individual ages. The increase in the expression levels of NAMPT2 and NANOG indicates that the addition of the supernatant of the culture solution to the skin cells rejuvenated the skin cells. In addition, NANOG is known to be a reprogramming factor. The increase in the expression level of NANOG indicates that the addition of the supernatant of the culture solution to the skin cells caused reprogramming and reprogramming of the cells, resulting in rejuvenation of the cells. In addition, it is shown that the supernatant of the culture solution has an anti-individual aging effect.

[0239] In addition, as shown in FIG. 67, it was shown that the expression level of TERT increased by adding the culture supernatant of Example 7 to skin cells. TERT enhances the expression of telomere elongation enzyme. Therefore, this result indicates that the culture supernatant can be used as a telomere elongation agent and a telomere shortening inhibitor.

[0240] Furthermore, as shown in Figures 68 and 69, it was shown that the expression levels of p53 and p16 decreased by adding the supernatant of the culture solution according to Example 7 to skin cells. p53 and p16 are senescence markers, and it is known that the expression levels of p53 and p16 increase when cells divide. The decrease in the expression levels of p53 and p16 indicates that the addition of the supernatant of the culture solution to the skin cells rejuvenated the skin cells. It also indicates that the supernatant of the culture solution has an anti-senescence effect and an anti-cellular aging effect.

[0241] (Example 9: Expression of rejuvenation genes in vivo) Eleven male and female rats aged 6 weeks were prepared. 5 mL of the supernatant of the culture solution according to Example 7 per kg of body weight was injected into the tail veins of six male rats and five female rats once a day for 14 days. In addition, as a control, 5 mL of saline per kg of body weight was injected into the tail veins of six male rats and five female rats once a day for 14 days. Two weeks after the first injection, fibroblasts were collected from each rat, and the expression levels of Manf, Gdf11, and Has2 in the fibroblasts were analyzed. As a result, as shown in Figures 70, 71, and 72, the expression levels of individual aging inhibitors such as Manf and Gdf11, and Has2 were increased in the skin tissue of rats that had been intravenously injected with the supernatant of the culture solution according to Example 7, compared to the control. This indicates that the aging of the individual rats was inhibited and the rats were rejuvenated.

[0242] Example 10: Expression of hyaluronic acid-related genes Fibroblasts were cultured in the same manner as in Example 8, and the supernatant of the culture solution was added. The RNA expression levels of hyaluronic acid producing enzymes HAS1, HAS2, and HAS3, and hyaluronic acid decomposition enzymes HYAL1 and HYAL3 were analyzed in the same manner as in Example 8. As a result, as shown in Figures 73, 74, and 75, the expression levels of hyaluronic acid producing enzymes HAS1, HAS2, and HAS3 were increased in fibroblasts to which the supernatant of the culture solution of Example 7 was added, compared to the control. In addition, as shown in Figures 76 and 77, the expression levels of hyaluronic acid decomposition enzymes HYAL1 and HYAL3 were decreased in fibroblasts to which the supernatant of the culture solution of Example 7 was added, compared to the control. This indicates that the supernatant of the culture solution can be used as an extracellular matrix production promoter, an extracellular matrix decomposition inhibitor, a rejuvenating agent, and an inhibitor of aging changes.

Claims

1. A lipolytic agent comprising stem cells, a stem cell extract, a stem cell culture medium, a culture supernatant during stem cell induction, a culture supernatant of stem cells, or a secretion product of stem cells.

2. An anti-obesity drug comprising stem cells, a stem cell extract, a stem cell culture medium, a culture medium supernatant during stem cell induction, a culture medium supernatant of stem cells, or a secretion product of stem cells.

3. An obesity improving drug comprising stem cells, a stem cell extract, a stem cell culture medium, a culture medium supernatant during stem cell induction, a culture medium supernatant of stem cells, or a secretion product of stem cells.

4. A dietary supplement comprising stem cells, a stem cell extract, a stem cell culture medium, a supernatant of a culture medium during stem cell induction, a supernatant of a culture medium of stem cells, or a secretion product of stem cells.

5. A drug for treating or preventing diabetes, comprising stem cells, a stem cell extract, a stem cell culture medium, a supernatant of a culture medium during stem cell induction, a supernatant of a culture medium of stem cells, or a secretion product of stem cells.

6. A muscle formation promoter comprising stem cells, a stem cell extract, a stem cell medium, a culture supernatant during stem cell induction, a culture supernatant of a stem cell, or a secretion product of a stem cell.

7. A muscle mass maintenance agent comprising stem cells, a stem cell extract, a stem cell medium, a culture supernatant during stem cell induction, a culture supernatant of stem cells, or a secretion product of stem cells.

8. A muscle mass enhancer comprising stem cells, a stem cell extract, a stem cell culture medium, a culture supernatant during stem cell induction, a culture supernatant of stem cells, or a secretion product of stem cells.

9. A prostacyclin production promoter comprising stem cells, a stem cell extract, a stem cell culture medium, a culture supernatant during stem cell induction, a culture supernatant of a stem cell, or a secretion product of a stem cell.

10. A platelet aggregation inhibitor comprising stem cells, a stem cell extract, a stem cell culture medium, a supernatant of a culture medium during induction of stem cells, a supernatant of a culture medium of stem cells, or a secretion product of stem cells.

11. A vasodilator comprising stem cells, a stem cell extract, a stem cell culture medium, a supernatant of a culture medium during stem cell induction, a supernatant of a culture medium of stem cells, or a secretion product of stem cells.

12. A drug for treating or preventing hypertension, comprising stem cells, a stem cell extract, a stem cell culture medium, a culture medium supernatant during stem cell induction, a culture medium supernatant of stem cells, or a secretion product of stem cells.

13. 1. A therapeutic or preventive agent for arteriosclerosis, comprising stem cells, a stem cell extract, a stem cell culture medium, a culture medium supernatant during stem cell induction, a culture medium supernatant of stem cells, or a secretion product of stem cells.

14. An agent for promoting extracellular matrix production by chondrocytes, comprising stem cells, a stem cell extract, a stem cell medium, a culture medium supernatant during stem cell induction, a culture medium supernatant of stem cells, or a secretion product of stem cells.

15. A chondrocyte protective or proliferation promoting agent comprising stem cells, a stem cell extract, a stem cell medium, a culture medium supernatant during stem cell induction, a culture medium supernatant of stem cells, or a secretion product of stem cells.

16. A drug for treating or preventing wounds in cartilage tissue, comprising stem cells, a stem cell extract, a stem cell culture medium, a supernatant of a culture medium during induction of stem cells, a supernatant of a culture medium of stem cells, or a secretion product of stem cells.

17. A drug for treating or preventing joint pain, comprising stem cells, a stem cell extract, a stem cell culture medium, a culture medium supernatant during stem cell induction, a culture medium supernatant of stem cells, or a secretion product of stem cells.

18. An agent for improving the survival rate of chondrocytes, comprising stem cells, a stem cell extract, a stem cell medium, a culture medium supernatant during stem cell induction, a culture medium supernatant of stem cells, or a secretion product of stem cells.

19. A painkiller comprising stem cells, a stem cell extract, a stem cell culture medium, a supernatant of a culture medium during stem cell induction, a supernatant of a culture medium of stem cells, or a secretion product of stem cells.

20. A cartilage regeneration agent comprising stem cells, a stem cell extract, a stem cell medium, a culture medium supernatant during stem cell induction, a culture medium supernatant of stem cells, or a secretion product of stem cells.

21. 1. A drug for treating, improving or preventing a joint, a joint disease or arthritis, comprising stem cells, a stem cell extract, a stem cell culture medium, a culture medium supernatant during stem cell induction, a culture medium supernatant of a stem cell, or a secretion product of a stem cell.

22. An agent for suppressing or alleviating inflammation, comprising stem cells, a stem cell extract, a stem cell culture medium, a culture medium supernatant during stem cell induction, a culture medium supernatant of stem cells, or a secretion product of stem cells.

23. An anti-aging agent comprising stem cells, a stem cell extract, a stem cell culture medium, a culture supernatant during stem cell induction, a culture supernatant of stem cells, or a secretion product of stem cells.

24. A rejuvenation agent comprising stem cells, a stem cell extract, a stem cell culture medium, a supernatant of a culture medium during stem cell induction, a supernatant of a culture medium of stem cells, or a secretion product of stem cells.

25. An agent for amplifying expression of a rejuvenation factor or a rejuvenation-related factor, comprising stem cells, a stem cell extract, a stem cell culture medium, a culture medium supernatant during stem cell induction, a culture medium supernatant of a stem cell, or a secretion product of a stem cell.

26. An agent for suppressing the expression of an aging marker, comprising stem cells, a stem cell extract, a stem cell medium, a culture supernatant during stem cell induction, a culture supernatant of a stem cell medium, or a secretion product of a stem cell.

27. An agent for promoting the production of extracellular matrix, comprising stem cells, a stem cell extract, a stem cell medium, a culture medium supernatant during stem cell induction, a culture medium supernatant of stem cells, or a secretion product of stem cells.

28. A telomere elongation agent or a telomere shortening inhibitor, comprising a stem cell, a stem cell extract, a stem cell medium, a culture supernatant during stem cell induction, a culture supernatant of a stem cell, or a secretion product of a stem cell.

29. A cell initialization agent, a cell de-differentiation agent, or a cell reprogramming agent, comprising stem cells, a stem cell extract, a stem cell medium, a culture medium supernatant during stem cell induction, a stem cell culture medium supernatant, or a stem cell secretion product.

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