Method for preparing mesenchymal stem cells by introducing peptides and pharmaceutical composition for treatment

Abstract

The present invention relates to a method for preparing a function-enhanced mesenchymal stem cell by introducing a peptide, and a pharmaceutical composition using the function-enhanced mesenchymal stem cell.

Description

Technical Field

[0001] This disclosure relates to a method for preparing functionally enhanced mesenchymal stem cells by introducing peptides, and a pharmaceutical composition using the functionally enhanced mesenchymal stem cells. Background Technology

[0002] Mesenchymal stem cells (MSCs) are stem cells found in cartilage, bone tissue, adipose tissue, bone marrow matrix, etc., and differentiate from the mesoderm formed from the division of a fertilized egg. MSCs are cells that maintain stemness and self-renewal, and possess the ability to differentiate into various mesenchymal tissues. MSCs can be extracted from bone marrow, adipose tissue, umbilical cord blood, synovium, bone tissue, infrapatellar fat pad, placenta, etc. MSCs possess immunomodulatory capabilities, inhibiting the activation and proliferation of T lymphocytes and B lymphocytes, suppressing natural killer (NK) cell activity, and regulating the function of dendritic cells and macrophages, making them suitable for allogeneic and xenograft transplantation. Given the potential of MSCs, extensive research is underway, including their application as cell-based therapies.

[0003] In research on mesenchymal stem cells, there have been studies aimed at overexpressing specific factors and using them as therapeutic compositions. Attempts have been made to treat diseases by overexpressing PEDF in mesenchymal stem cells (Patent Document 002), and attempts have been made to treat diseases by overexpressing PRL-1 in mesenchymal stem cells (Patent Documents 003 to 005).

[0004] Protein tyrosine phosphatase type IV A1 (PTP4A1), also known as regenerative liver phosphatase-1 (PRL-1), belongs to the group of bispecific protein tyrosine phosphatases capable of removing phosphorylation from both tyrosine and serine / threonine residues among 109 mammalian protein tyrosine phosphatases. PTP4A1 is known to influence cell growth and liver regeneration. One method exists that uses an electroporation technique called AMAXA to deliver a plasmid containing the PRL-1 gene into cells to induce PRL-1 overexpression in mesenchymal stem cells (Patent Documents 003 to 005). However, safety issues may arise when using the electroporation method due to the introduction of the plasmid. Summary of the Invention

[0005] Technical issues

[0006] In this disclosure, a method is provided for culturing mesenchymal stem cells in a culture medium containing a peptide that consists of or contains the core amino acid sequence of SEQ ID NO:1.

[0007] In this disclosure, mesenchymal stem cells cultured in a culture medium containing a peptide that consists of or contains the core amino acid sequence of SEQ ID NO:1 are provided.

[0008] Solution

[0009] To achieve the above objective, a method for preparing mesenchymal stem cells is provided, comprising: (a) providing mesenchymal stem cells; and (b) culturing the mesenchymal stem cells in a culture medium containing PRL-1 peptide.

[0010] In this disclosure, the method may further include: prior to step (b), culturing mesenchymal stem cells in a culture medium that does not contain PRL-1 peptide.

[0011] In this disclosure, the method may further include: after step (b), culturing mesenchymal stem cells in a culture medium that does not contain PRL-1 peptide.

[0012] In this disclosure, the method may further include: after step (b), culturing mesenchymal stem cells in a culture medium containing the PRL-1 peptide.

[0013] In this disclosure, mesenchymal stem cells may be placental-derived mesenchymal stem cells.

[0014] In this disclosure, mesenchymal stem cells can be induced to overexpress the PRL-1 gene via the PRL-1 peptide.

[0015] In this disclosure, the concentration of PRL-1 peptide can be from 5 to 500 pg / ml.

[0016] In this disclosure, mesenchymal stem cells can be induced to express at least one surface antigen selected from the group consisting of CD34, CD105, HLA-DR, and HLA-G via PRL-1 peptide.

[0017] In this disclosure, the expression of Col 1 can be reduced via the PRL-1 peptide.

[0018] In this disclosure, the expression of at least one gene selected from the group consisting of albumin, cyclin D1, and HNF1a can be increased by the PRL-1 peptide.

[0019] To achieve the above objectives, a functionally enhanced mesenchymal stem cell is provided, which is prepared by a method for preparing mesenchymal stem cells, the method comprising: (a) providing mesenchymal stem cells; and (b) culturing the mesenchymal stem cells in a culture medium containing PRL-1 peptide.

[0020] In this disclosure, a composition for the prevention or treatment of abnormal lipogenesis and metabolic disorders is provided, comprising functionally enhanced mesenchymal stem cells.

[0021] In this disclosure, a composition for the prevention or treatment of liver diseases is provided, comprising functionally enhanced mesenchymal stem cells.

[0022] Beneficial effects

[0023] According to this disclosure, overexpression of PRL-1 in mesenchymal stem cells can be induced without transduction.

[0024] The enhanced mesenchymal stem cells disclosed herein can be used to prevent or treat abnormal adipogenesis, metabolic disorders, liver diseases, etc. Attached Figure Description

[0025] Figure 1 The study showed that the expression level of the PRL-1 gene increased in mesenchymal stem cells after the introduction of the PRL-1 peptide.

[0026] Figure 2 The study showed that the level of PRL-1 protein increased in mesenchymal stem cells after the introduction of the PRL-1 peptide.

[0027] Figure 3 The results obtained by measuring the doubling time of mesenchymal stem cells after the introduction of the PRL-1 peptide are shown.

[0028] Figure 4 The results obtained by measuring PRL-1 protein levels in mesenchymal stem cells after the introduction of the PRL-1 peptide are shown.

[0029] Figure 5 The results show the comparison of PRL-1 gene expression levels in mesenchymal stem cells after the introduction of the PRL-1 peptide.

[0030] Figure 6 The results obtained by comparing PRL-1 protein levels in mesenchymal stem cells after the introduction of the PRL-1 peptide are shown.

[0031] Figure 7 The results show the comparison of OCT4 expression levels in mesenchymal stem cells after the introduction of the PRL-1 peptide.

[0032] Figure 8 The results obtained by flow cytometry of mesenchymal stem cells after the introduction of the PRL-1 peptide are shown.

[0033] Figure 9 The results obtained by flow cytometry of mesenchymal stem cells after the introduction of the PRL-1 peptide are shown.

[0034] Figure 10The results obtained in identifying the osteogenic differentiation potential of mesenchymal stem cells after the introduction of the PRL-1 peptide are shown.

[0035] Figure 11 The results obtained in identifying the adipogenic differentiation potential of mesenchymal stem cells after the introduction of the PRL-1 peptide are shown.

[0036] Figure 12 The results obtained in identifying the chondrogenic differentiation potential of mesenchymal stem cells after the introduction of the PRL-1 peptide are shown.

[0037] Figure 13 The results obtained in identifying the cytokine secretion potential of mesenchymal stem cells after the introduction of the PRL-1 peptide are shown.

[0038] Figure 14 The results show the identification of the antifibrotic effect of mesenchymal stem cells on hepatic stellate cell lines after the introduction of the PRL-1 peptide.

[0039] Figure 15 The results show the effects of introducing the PRL-1 peptide on hepatocyte regeneration of mesenchymal stem cells. Detailed Implementation

[0040] This disclosure can be fully implemented by the following description. The following description should be understood as illustrating specific embodiments of this disclosure, and this disclosure is not necessarily limited thereto. Furthermore, it should be understood that the accompanying drawings are provided for ease of understanding only, and this disclosure is not limited thereto.

[0041] The embodiments of this disclosure can be modified in various ways and can be implemented in various forms. Therefore, it should be understood that this disclosure includes all modifications, equivalents, and alternatives that fall within the technical concept and scope of this disclosure.

[0042] Mesenchymal stem cells (MSCs) can refer to cells that maintain self-renewal and stemness and are capable of differentiating into various mesenchymal tissues, and can include mesenchymal stem cells from animals including mammals (e.g., humans). Furthermore, mesenchymal stem cells can be derived from umbilical cord, umbilical cord blood, bone marrow, placental, or adipose tissue. Placental-derived mesenchymal stem cells can be derived from various sources constituting the placenta, such as amniotic epithelial cells, or tissues such as the amnion, trophoblast, or chorion. Preferably, placental-derived mesenchymal stem cells can be mesenchymal stem cells derived from the placental chorionic plate, and more preferably, they can be mesenchymal stem cells derived from the chorionic plate membrane. The isolation of mesenchymal stem cells can be performed by methods readily apparent to those skilled in the art.

[0043] Regenerative liver phosphatase-1 (PRL-1) may include PRL-1 derived from vertebrates, such as mammals, fish, amphibians, birds, or reptiles, including humans. Furthermore, it is understood that PRL-1 includes PRL-1 precursors.

[0044] PRL-1 can be prepared as a fusion protein. In the preparation of this fusion protein, the polynucleotide encoding PRL-1 can be in-frame linked to a polynucleotide encoding another protein or peptide. For this purpose, techniques known in the art can be utilized. For the peptide fused with PRL-1, well-known peptides can be used, such as FLAG (Hopp, TP et al., BioTechnology (1988) 6, 1204-1210), 6xHis residues consisting of six histidines (His), 10xHis, influenza hemagglutinin (HA), human c-myc fragment, VSV-GP fragment, p18HIV fragment, T7 tag, HSV tag, E tag, SV40T antigen fragment, lck tag, α-tubulin fragment, B tag, and protein C fragment. In addition, to generate the fusion protein, PRL-1 can be linked to glutathione S-transferase (GST), influenza hemagglutinin (HA), immunoglobulin constant region, β-galactosidase, maltose-binding protein (MBP), etc.

[0045] As used in this article, the term "via PRL-1 peptide" refers to the effect of the presence of PRL-1 peptide on cells. The process by which cells are affected can involve the PRL-1 peptide exerting its function within the cell.

[0046] Overexpression of the PRL-1 gene in mesenchymal stem cells can be induced by the PRL-1 peptide. Overexpression refers to an increase in the expression level of the PRL-1 gene compared to the absence of the PRL-1 peptide.

[0047] PRL-1 peptide can induce the expression of at least one surface antigen selected from the group consisting of CD34, CD105, HLA-DR, and HLA-G in mesenchymal stem cells. PRL-1 peptide can also induce the reduction of Col 1 expression in mesenchymal stem cells. Furthermore, PRL-1 peptide can induce the increase of at least one gene selected from the group consisting of albumin, cyclin D1, and HNF1a in mesenchymal stem cells.

[0048] The culture medium may include α-MEM or DMEM / F12 medium. The medium may also contain other substances. These other substances may include penicillin-streptomycin, heparin, and fibroblast growth factor.

[0049] Disorders of dyslipidemia and metabolism can refer to diseases caused by abnormal lipid metabolism. Examples of disorders of dyslipidemia and metabolism may include obesity, diabetes, dyslipidemia, metabolic diseases, hypertension, thyroid eye disease, or degenerative diseases associated with disorders of dyslipidemia and metabolism.

[0050] In addition, metabolic disorders can include menopausal disorders based on metabolic diseases, ovarian dysfunction (insufficient ovarian function and polycystic ovary syndrome), and metabolic osteoporosis.

[0051] Liver diseases can include liver damage, liver dysfunction, hepatitis, hepatotoxicity, cholestasis, fatty liver, cirrhosis, hepatic ischemia, alcoholic liver disease, liver abscess, hepatic coma, liver atrophy, or liver cancer. A state in which liver disease persists and the liver is unable to perform normal liver functions is termed "liver dysfunction." Patients with liver dysfunction may exhibit significantly increased expression of fibrosis-related genes compared to healthy individuals; and / or significantly decreased levels of cyclin D1, HNF1a, and albumin (ALB) compared to healthy individuals.

[0052] This disclosure will be described in more detail through the following embodiments. However, these embodiments are only used to illustrate specific implementations of this disclosure, and this disclosure is not limited thereto.

[0053] 1. Preparation examples, implementation examples, and comparative examples

[0054] Preparation Example 1. Isolation and Culture of Placental Derived Mesenchymal Stem Cells

[0055] After obtaining informed consent from a healthy mother who has received a fully explained explanation, the umbilical cord was isolated from placental tissue collected during normal delivery. The isolated tissue (chorionic amnion) was placed in a 50 ml tube, and DPBS was added to remove excess blood. Then, using a sterile glass slide, the upper part of the chorionic amnion was scraped in 20 ml of enzyme solution I (1 mg / ml collagenase type I, 2 mg / ml trypsin, 20 mg / ml DNase I, 1.2 U / ml dispersant, x1 PS of HBSS solution), and the recovered suspension was collected to one side. 10 ml of enzyme solution I was added to this and mixed thoroughly. The enzymatic reaction was then carried out in two rounds at 37°C for 15 minutes each, thereby isolating stem cells from the tissue. The separated cell suspension was centrifuged, and the separated cells were cultured in a 37°C incubator under 20% O2 and 5% CO2 conditions in DMEM / F12 medium supplemented with 10% fetal bovine serum, 1% penicillin-streptomycin, 1 μg / ml heparin and 25 ng / ml fibroblast growth factor-4 (FGF-4).

[0056] Preparation Example 2. Preparation of PRL-1 peptide solution

[0057] A peptide solution was prepared by adding recombinant human PTP4A1 peptide PRL-1 (NOVUS, NBP1-50946-0.1 mg) to α-MEM medium to achieve a PRL-1 peptide concentration of 1 mg / ml. The amino acid sequence of the human PRL-1 peptide is shown in Table 1.

[0058] [Table 1]

[0059]

[0060] Example 1. Culture of mesenchymal stem cells using peptide solution (single cycle)

[0061] (1) 5th generation single time

[0062] The mesenchymal stem cells cultured according to Preparation Example 1 were passaged four times, with the culture medium changed at 4-5 day intervals. Then, for the fifth passage, the cells were cultured for 120 hours in a medium supplemented with the peptide solution from Preparation Example 2 to achieve a PRL-1 peptide concentration of 500 pg / ml.

[0063] (2) 6th generation single time

[0064] The mesenchymal stem cells cultured according to Preparation Example 1 were passaged four times, with the culture medium changed at 4-5 day intervals. Then, for the fifth passage, the cells were cultured in a medium supplemented with the peptide solution from Preparation Example 2, so that the concentration of PRL-1 peptide was 500 pg / ml. For the sixth passage, the cells were cultured in a medium without the added peptide solution.

[0065] (3) 7th generation single time

[0066] The mesenchymal stem cells cultured according to Preparation Example 1 were passaged four times, with the culture medium changed every 4 to 5 days. Then, for the fifth passage, the cells were cultured in a medium supplemented with the peptide solution from Preparation Example 2, so that the concentration of PRL-1 peptide was 500 pg / ml. For the sixth and seventh passages, the cells were cultured in a medium without the added peptide solution.

[0067] Example 2. Culture of mesenchymal stem cells using peptide solution (multiple times)

[0068] (1) 5th generation multiple times

[0069] The mesenchymal stem cells cultured according to Preparation Example 1 were passaged four times, with the culture medium changed at 4-5 day intervals. Then, for the fifth passage, the cells were cultured in a medium supplemented with the peptide solution from Preparation Example 2, so that the concentration of PRL-1 peptide was 500 pg / ml.

[0070] (2) 6th generation multiple times

[0071] The mesenchymal stem cells cultured according to Preparation Example 1 were passaged four times, with the culture medium changed at 4-5 day intervals. Then, for the fifth and sixth passages, the cells were cultured in a medium supplemented with the peptide solution from Preparation Example 2, so that the concentration of PRL-1 peptide was 500 pg / ml.

[0072] (3) 7th generation multiple times

[0073] The mesenchymal stem cells cultured according to Preparation Example 1 were passaged four times, with the culture medium changed at 4-5 day intervals. Then, for the fifth to seventh passages, the cells were cultured in a medium supplemented with the peptide solution from Preparation Example 2, so that the concentration of PRL-1 peptide was 500 pg / ml.

[0074] Comparative Example 1: Negative Control (Initial)

[0075] The mesenchymal stem cells cultured according to Preparation Example 1 were passaged 5 to 7 times, with the culture medium changed at 4 to 5-day intervals, and used as a negative control.

[0076] Comparative Example 2. Preparation of a positive control using electroporation (AMAXA)

[0077] The mesenchymal stem cells cultured according to Preparation Example 1 were passaged 4 times, with the culture medium being changed at intervals of 4 to 5 days.

[0078] Subsequently, placental chorionic lamina-derived mesenchymal stem cells (CP-MSCs) were cultured in α-MEM medium supplemented with 10% fetal bovine serum (FBS), 1% penicillin-streptomycin, 25 ng / ml fibroblast growth factor-4 (FGF-4), and 1 μg / ml heparin in a CO2 incubator at 37°C. Cells were washed with phosphate-buffered saline (PBS) and then treated with trypsin at 37°C for 2 minutes to detach the cells. Dedetached cells were collected using PBS and centrifuged at 1200 rpm for 5 minutes. The resulting pellet was resuspended in 1 ml of medium, and cells were counted using a hemocytometer. The recovered cells were centrifuged at 200 g for 10 minutes and resuspended in Nucleofector solution to a concentration of 5 × 10⁻⁶. 5Cells were added at 100 μl, followed by the addition of 2 μg of a DNA plasmid containing PRL-1. Cells were transferred to cuvettes and placed in a Nucleofector device, and program U-23 was run. After the program was complete, cells were rapidly transferred to culture dishes containing fresh medium and stabilized in a CO2 incubator at 37°C. After 4 hours, the medium was removed from the culture dishes using a pipette. The adherent cells were then cultured in α-MEM medium supplemented with 1.5 mg / ml neomycin, 10% fetal bovine serum (FBS), 1% penicillin-streptomycin, 25 ng / ml fibroblast growth factor-4 (FGF-4), and 1 μg / ml heparin to obtain placental chorionic lamina-derived mesenchymal stem cells transduced with the PRL-1 gene.

[0079] 2. Analysis of preparation examples, implementation examples, and comparative examples

[0080] (1) Expression of PRL-1 gene and synthesis of PRL-1 protein after introduction of PRL-1 peptide.

[0081] Mesenchymal stem cells from Example 1 were prepared by treating the peptide solution from Preparation Example 2 at concentrations of 5 pg / ml, 50 pg / ml, and 500 pg / ml. Figure 1 As shown, treatment with peptide solution for 24 and 72 hours resulted in increased expression levels of PRL-1 mRNA. Furthermore, as... Figure 2 As shown, it displays the results of protein blotting and protein level measurements, revealing that PRL-1 protein levels increased after treatment with peptide solution for 24 and 72 hours.

[0082] (2) Comparison between the subcultured examples and the comparative examples

[0083] The cell doubling time was compared in Example 1 (single passage), Example 2 (multiple passages), and Comparative Example 1 (initial) (all passaged 5 to 7 times). Figure 3 As shown, Examples 1 and 2 did not show a significant difference in doubling time compared to the initial generation without any treatment.

[0084] PRL-1 levels were analyzed by ELISA for Example 1 (single), Example 2 (multiple), and Comparative Example 1 (initial) (all passaged 5 to 7 times), and for Comparative Example 2 (AMAXA) using electroporation. Figure 4 As shown, the PRL-1 protein was synthesized.

[0085] The PRL-1 mRNA expression levels and PRL-1 protein levels of Example 1 (single), Example 2 (multiple), Comparative Example 1 (initial) (all passaged 5 to 7 times), and Comparative Example 2 (AMAXA) were compared. Figure 5As shown, low PRL-1 mRNA expression levels were observed in Examples 1 and 2 after 6 to 7 passages; however, as Figure 6 As shown, there was no significant difference in PRL-1 protein levels.

[0086] The expression levels of the undifferentiated marker OCT4 in Example 1 (single passage), Example 2 (multiple passages), and Comparative Example 1 (initial passage) (all passaged 5 to 7 times) were compared with those in Comparative Example 2 (AMAXA) passaged 9 to 11 times. The results are as follows: Figure 7 As shown.

[0087] Figure 8 and Figure 9 The results obtained by flow cytometry (FACS) of Example 1 (single passage), Example 2 (multiple passages), and Comparative Example 1 (initial) (all passaged 5 to 7 times) are shown. Figure 8 and Figure 9 As shown, the cells were found to be negative for hematopoietic markers (CD34 and HLA-DR) and positive for non-hematopoietic markers (CD13, CD90, CD105, HLA-ABC, and HLA-G). Furthermore, Examples 1 and 2 showed no significant differences compared to Comparative Example 1.

[0088] (3) Identification of differentiation potential of the examples and comparative examples

[0089] Differentiation was induced in Examples 1 and 2 to identify their differentiation potential as mesenchymal stem cells.

[0090] To identify the osteogenic differentiation potential of Examples 1 and 2, von Kossa staining was performed to detect mineralization in the tissues based on calcium, potassium, etc. Figure 10 As can be seen, staining was observed in both the examples and comparative examples. Furthermore, as... Figure 10 As shown, compared with undifferentiated cells (Undiff), the examples and comparative examples showed increased expression of osteoblast-specific genes (osteocalcin), indicating that they have osteogenic differentiation potential.

[0091] To identify the adipogenic differentiation potential of Examples 1 and 2, Oil Red O staining was performed to stain the lipids. Figure 11 As shown, staining was observed in both the examples and comparative examples, and there was no significant difference in staining levels between them. Furthermore, as... Figure 11 As shown, compared with undifferentiated cells (Undiff), the examples and comparative examples exhibited increased expression of adipocyte-specific genes (lipoproteins), indicating that they have adipogenic differentiation potential.

[0092] To identify the chondrogenic differentiation potential of Examples 1 and 2, alcian blue staining was performed on the cartilage. Figure 12As can be seen, compared with undifferentiated cells (Undiff), Examples 1 and 2 and Comparative Example 2 showed low expression levels of Sox9 and ColII genes associated with chondrogenic differentiation potential; this result indicates that their chondrogenic differentiation potential is limited. However, Example 2 was found to show higher expression levels of Sox9 and ColII genes than Example 1 and Comparative Example 2.

[0093] (4) Identification of cytokine secretion potential in the examples and comparative examples

[0094] To identify the cytokine secretion potential of placental mesenchymal stem cells after the introduction of PRL-1 peptide, the cytokine secretion levels of Comparative Example 1 (initial), Example 1 (single), and Example 2 (multiple) were compared.

[0095] Comparative Example 1, Example 1, and Example 2, all passaged seven times, were cultured for 24 hours. Then, the culture medium was replaced with FBS-free medium, and the cells were cultured for another 24 hours. The cytokine secretion potential was then assessed using this medium. Figure 13 As shown, the introduction of the PRL-1 peptide resulted in increased expression of regenerative potential-related factors (growth factors), immune-related factors, metabolic-related factors, and angiogenesis-related factors in placental-derived mesenchymal stem cells compared to the initial sample. Example 1 was found to exhibit higher cytokine secretion levels than Example 2.

[0096] (5) Identification of the antifibrotic effect of hepatic stellate cell line

[0097] Hepatic stellate cell lines were cultured in T-HSC medium for 24 hours, and then treated with 2 ng / ml TGF-β for 24 hours to induce fibrosis. The hepatic stellate cell lines were then treated with the methods described in Example 1, Example 2, Comparative Example 1, and Comparative Example 2, and cultured for 24 hours to identify their anti-fibrotic effects. Figure 14 As shown, the anti-fibrotic effect was identified by the expression level and protein amount of Col 1.

[0098] (6) Identification of the effect on hepatocyte regeneration

[0099] Hepatocyte lines were cultured in WB-F344 medium for 24 hours, followed by chemical treatment (LCA) for 48 hours to induce cell damage. Then, the hepatocyte lines were treated with the methods described in Example 1, Example 2, Comparative Example 1, and Comparative Example 2, and cultured for 24 hours to assess their cell regeneration effects. Figure 15 As shown, the increased expression levels of albumin (ALB), cyclin D1, and HNF1a secreted by normal hepatocytes were used to identify the hepatocyte regeneration effect.

Claims

1. A method for preparing mesenchymal stem cells, comprising: (a) Providing mesenchymal stem cells; as well as (b) The mesenchymal stem cells were cultured in a medium containing regenerating liver phosphatase-1 (PRL-1) peptide.

2. The method according to claim 1, further comprising: Prior to step (b), the mesenchymal stem cells were cultured in a medium that did not contain the PRL-1 peptide.

3. The method according to claim 1 or 2, further comprising: After step (b), the mesenchymal stem cells are cultured in a medium that does not contain PRL-1 peptide.

4. The method according to claim 1 or 2, further comprising: Following step (b), the mesenchymal stem cells are cultured in a medium containing the PRL-1 peptide.

5. The method according to claim 1, wherein the mesenchymal stem cells are placental-derived mesenchymal stem cells.

6. The method of claim 1, wherein the mesenchymal stem cells are induced to overexpress the PRL-1 gene by a peptide comprising the core amino acid sequence of PRL-1.

7. The method according to claim 1, wherein the concentration of the PRL-1 peptide in the culture medium is from 1 pg / ml to 1000 pg / ml.

8. The method of claim 1, wherein the mesenchymal stem cells are induced to express at least one surface antigen selected from the group consisting of CD34, CD105, HLA-DR, and HLA-G by the PRL-1 peptide.

9. A functionally enhanced mesenchymal stem cell, prepared by a method for preparing mesenchymal stem cells, the method comprising: (a) Providing mesenchymal stem cells; as well as (b) The mesenchymal stem cells are cultured in a culture medium containing peptides, the peptides containing the core amino acid sequence of PRL-1.

10. A composition for the prevention or treatment of abnormal lipogenesis, metabolic disorders and degenerative diseases, comprising the functionally enhanced mesenchymal stem cells of claim 9.

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