JP2019214514A - Culture supernatant formulation for increasing antibody productivity - Google Patents

Abstract

To provide formulations for increasing antibody productivity.SOLUTION: Provided is a culture supernatant formulation containing a culture supernatant of a mesenchymal stem cell as an active ingredient to enhance antibody productivity. An example of the mesenchymal stem cell is a human mesenchymal stem cell, and more preferably, the mesenchymal stem cell is cultured using a serum-free medium. This preparation is used, for example, to enhance the IgA or IgG antibody productivity of an administered subject.SELECTED DRAWING: Figure 1

Description

  The present invention relates to a culture supernatant preparation for increasing the antibody productivity of a subject.

  Antibodies are extremely important for host defense. Production of secretory IgA antibodies in the intestinal tract, which is a tissue that comes into contact with foreign substances, and production of IgG antibodies that are frequently present in blood play a large role in controlling various diseases including infectious diseases. However, it is known that antibody production ability decreases with age. This is considered to be one reason, for example, that the number of deaths due to pneumonia is high in the elderly.

  As mentioned above, the loss of antibody production due to aging increases the risk of developing an infection. It is also a very serious problem because it prevents the antibody from being boosted by the vaccine. However, at this time, there is no effective means to increase the antibody production capacity that has declined due to aging. Therefore, development of agents that enhance systemic immunity and intestinal immunity has been desired.

  By the way, mesenchymal stem cells (MSCs) isolated from adipose tissue, umbilical cord, bone marrow, dental pulp, etc., adhere to plastic culture dishes, grow by culture, and grow into osteoblasts, chondrocytes, adipocytes, etc. It has the property of having the ability to differentiate. These cells are known to act directly on the site of the disease or by secreting various cytokines, thereby acting on wound healing, angiogenesis, anti-inflammatory, immunosuppression and the like.

  The present inventors have examined the possibility that a culture supernatant containing secretions of MSCs can be applied to enhance antibody production.

International Patent Application WO 2009102050 pamphlet

Corcione, A. et al., "Human mesenchymal stem cells modulate B-cell functions" Blood (The American Society of Hematology) 2006 Comoli, P. et al., "Human mesenchymal stem cells inhibit antibody production induced in vitro by allostimulation" Nephrology Dialysis Transplant (Oxford University Press) 2008 Saka, Y. et al. "Adipose-derived stromal cells cultured in a low-serum medium, but not bone marrow-derived stromal cells, impede xenoantibody production" Xenotransplantation (John Wiley & Sons A / S) 2011 Park, M.-J. et al., `` Adipose tissue-derived mesenchymal stem cells induce expansion of Interleukin-10 producing regulatory B cells and ameliorate autoimmunity in a murine model of systemic lupus erythematosus, '' Cell Transplantation (Cognizant Communication Corporation) 2014

  An object of the present invention is to provide an agent that enhances antibody production ability.

  An object of the present invention is to provide an agent for enhancing antibody production of a subject, comprising a culture supernatant of mesenchymal stem cells having a function of increasing the antibody production of a subject.

  As described above, mesenchymal stem cells secrete various factors and are effective in various diseases and wounds. So far, several academic reports have been published on the relationship between mesenchymal stem cells and immune-related cells. According to this, mesenchymal stem cells act to suppress the differentiation and proliferation of antibody-producing cells, suppress the maturation of antigen-presenting cells, and consequently suppress antibody production.

  However, the above results were obtained using mesenchymal stem cells cultured in a serum-containing medium. It is known that mesenchymal stem cells cultured in a serum-containing medium and a serum-free medium have different gene expression and secretory profiles.

  Therefore, the present inventors cultured mesenchymal stem cells using a serum-free medium instead of a serum-containing medium, and produced a culture supernatant preparation (culture supernatant) containing secretions of mesenchymal stem cells. Using this culture supernatant, the effect on antibody production ability was verified.

  A culture supernatant prepared using adipose tissue, umbilical cord, and dental pulp-derived mesenchymal stem cells is administered to aged mice (18 to 22 months old), and their antibody-producing ability is compared with that of mice without culture supernatant. did. Ovalbumin (OVA) and cholera toxin (CT) were used as antigens and were orally ingested.

  In the aged mice to which the culture supernatant was not administered, production of intestinal secretory IgA antibodies and IgG antibodies against OVA and CT was not observed or was extremely low. However, in the aged mice to which the culture supernatant was administered, the above-mentioned remarkable increase in antibody production was confirmed. (FIGS. 1-10).

  As described above, it was shown that the culture supernatant of mesenchymal stem cells produced using the serum-free medium enhances the antibody-producing ability. This is contrary to the results of previous studies, namely, that mesenchymal stem cells cultured in serum-containing medium suppress the activity of immune-related cells such as B cells and T cells and reduce antibody production. Is the result of

  The present invention relates to a culture supernatant preparation for enhancing antibody-producing ability, comprising a culture supernatant of mesenchymal stem cells as an active ingredient. A preferred form of this agent uses human mesenchymal stem cells as the mesenchymal stem cells. In a preferred form of this agent, the culture supernatant of mesenchymal stem cells is a culture supernatant obtained when culturing mesenchymal stem cells using a serum-free medium. A preferred form of this formulation is one that is used to enhance the ability of the administered subject to produce IgA or IgG antibodies. The agent of the present invention is effective as an infectious disease preventive agent, a vaccine enhancer, or an anti-cancer active agent because it enhances the ability of an administered subject to produce an IgA antibody or IgG antibody.

  According to the present invention, by culturing mesenchymal stem cells separated from a body tissue, a culture supernatant having a function of enhancing antibody production ability can be obtained. Therefore, according to the present invention, it is possible to provide a method for producing a culture supernatant having a function of enhancing antibody production ability, and an agent containing a culture supernatant having a function of enhancing antibody production ability as a component.

FIG. 1 is a graph showing that a culture supernatant of adipose tissue-derived mesenchymal stem cells cultured in a serum-free medium (BMS medium) of Biomimetics Synpathies Inc. increases antibody titer against OVA in aged mice. . The left shows the antibody titer of secretory IgA antibody, and the right shows the antibody titer of serum IgG antibody. FIG. 2 is a graph showing that the culture supernatant of adipose tissue-derived mesenchymal stem cells cultured in a serum-free medium (Lonza medium) of Lonza raises the antibody titer against OVA in aged mice. The left shows the antibody titer of secretory IgA antibody, and the right shows the antibody titer of serum IgG antibody. FIG. 3 is a graph showing that the culture supernatant of adipose tissue-derived mesenchymal stem cells cultured in a serum-free medium (thermo medium) of Thermo Fisher Scientific Inc. increases the antibody titer against OVA in aged mice. is there. The left shows the antibody titer of secretory IgA antibody, and the right shows the antibody titer of serum IgG antibody. FIG. 4 is a graph showing that a culture supernatant of adipose tissue-derived mesenchymal stem cells cultured in a BMS medium increases antibody titer to CT in aged mice. The left shows the antibody titer of secretory IgA antibody, and the right shows the antibody titer of serum IgG antibody. FIG. 5 is a graph showing that a culture supernatant of adipose tissue-derived mesenchymal stem cells cultured in Lonza medium increases antibody titer to CT in aged mice. The left shows the antibody titer of secretory IgA antibody, and the right shows the antibody titer of serum IgG antibody. FIG. 6 is a graph showing that a culture supernatant of adipose tissue-derived mesenchymal stem cells cultured in a thermo medium increases an antibody titer to CT in aged mice. The left shows the antibody titer of secretory IgA antibody, and the right shows the antibody titer of serum IgG antibody. FIG. 7 is a graph showing that umbilical cord-derived mesenchymal stem cells cultured in BMS medium increase the antibody titer against OVA in aged mice. The left shows a secretory IgA antibody, and the right shows a serum IgG antibody. FIG. 8 is a graph showing that pulp-derived mesenchymal stem cells cultured in BMS medium increase the antibody titer against OVA in aged mice. The left shows a secretory IgA antibody, and the right shows a serum IgG antibody. FIG. 9 is a graph showing that umbilical cord-derived mesenchymal stem cells cultured in BMS medium increase the antibody titer against CT in aged mice. The left shows a secretory IgA antibody, and the right shows a serum IgG antibody. FIG. 10 is a graph showing that dental pulp-derived mesenchymal stem cells cultured in BMS medium increase the antibody titer against CT in aged mice. The left shows a secretory IgA antibody, and the right shows a serum IgG antibody.

  Hereinafter, embodiments for carrying out the present invention will be described with reference to the drawings. The present invention is not limited to the embodiments described below, but includes those appropriately modified by those skilled in the art from the following embodiments.

  A method for producing a culture supernatant of mesenchymal stem cells will be described. The culture supernatant preparation of the present invention relates to a culture supernatant preparation for enhancing antibody-producing ability, comprising a culture supernatant of mesenchymal stem cells as an active ingredient. A preferred form of this agent uses human mesenchymal stem cells as the mesenchymal stem cells. A preferred form of this agent was obtained when the culture supernatant of mesenchymal stem cells was cultured using an inanimate culture method using a serum-free and animal-free extract-free medium. Culture supernatant. A preferred form of this formulation is one that is used to enhance the ability of the administered subject to produce IgA or IgG antibodies.

  Mesenchymal stem cells refer to somatic stem cells that can be differentiated into cells constituting various types of mesenchymal tissues. Examples of mesenchymal stem cells are mesenchymal stem cells of mammals (eg, humans), more preferably primate mesenchymal stem cells, and even more preferably human mesenchymal stem cells. Mesenchymal stem cells can also be present, for example, in bone marrow, fat, dental pulp and umbilical cord. The mesenchymal stem cells may be a cell population containing mesenchymal stem cells. The mesenchymal stem cells may have been subcultured.

A serum-free medium containing no autologous serum or fetal bovine serum (FBS) is used as a culture solution. A medium free of human or animal-derived components is more preferred. If necessary, a growth factor such as fibroblast growth factor (bFGF) or adrenomedullin may be added. The culture can be carried out under any conditions suitable for culturing mammalian cells. In general, the cells are cultured at 37 ° C. and 5% CO ₂ for several days, and the medium is replaced as necessary. Since mesenchymal stem cells have the property of proliferating by adhering to the culture substrate, they can be easily separated from hematopoietic stem cells that proliferate in suspension.

  The culture supernatant of mesenchymal stem cells may be a supernatant obtained by centrifuging the medium used in cell culture. Further, the supernatant may be appropriately purified. In addition, after using the method for producing mesenchymal stem cells described above, the medium was replaced with a general basic medium such as Dulbecco's MEM (registered trademark) medium, a commercially available medium, or a culture medium or buffer having a unique composition. Later, the supernatant may contain components secreted from mesenchymal stem cells.

  Examples of culture supernatants of mesenchymal stem cells include a supernatant obtained by solid-liquid separation of the culture supernatant by centrifugation, a processed supernatant obtained by removing water by lyophilization, and an evaporator. The product obtained by concentrating the culture supernatant under reduced pressure using a filter, the product obtained by concentrating the culture supernatant using an ultrafiltration membrane, or the like, or by solid-liquid separation of the culture supernatant using a filter It is a processed product obtained or a stock solution of a culture supernatant before the above-described treatment. In addition, for example, the supernatant obtained by culturing the mesenchymal stem cells of the present invention is centrifuged (for example, 1,000 × g, 10 minutes), and then fractionated with ammonium sulfate (for example, 65% saturated ammonium sulfate). May be suspended in an appropriate buffer, dialyzed, and filtered through a syringe filter (eg, 0.2 μm) to obtain a sterile culture supernatant. The collected culture supernatant can be used as it is or can be stored frozen and thawed at the time of use. Alternatively, a pharmaceutically acceptable carrier may be added, and the solution may be dispensed into a sterile container so as to have an easily handled liquid volume, for example, 0.2 ml or 0.5 ml. Furthermore, as a measure against the risk of infectious pathogens, the culture supernatant may be treated with a virus clearance filter or gamma irradiation.

  An agent containing a culture supernatant as an active ingredient is known as disclosed in, for example, JP-A-2013-18756, Japanese Patent No. 5139294, and Japanese Patent No. 5526320. Therefore, the agent containing the culture supernatant of the present invention can be produced by a known method. A preferred form of the agent of the present invention is used for enhancing the ability of an administered subject to produce an IgA antibody or IgG antibody. The agent of the present invention is effective as an infectious disease preventive agent, a vaccine enhancer, or an anti-cancer active agent because it enhances the ability of an administered subject to produce an IgA antibody or IgG antibody. An infectious disease preventive agent is an agent for preventing an administered subject from becoming infected. The infectious disease preventive agent may be a component of a health food for preventing infectious disease, or may be an approved drug. A vaccine enhancer is an agent that is administered together with or used in combination with a vaccine, and enhances the therapeutic effect of the vaccine. An anti-cancer active agent is an agent that is an anti-cancer agent, or that is administered with or used in combination with an anti-cancer agent to enhance the activity of the anti-cancer agent.

  As the dosage form of the culture supernatant according to the present invention, both a liquid preparation and a solid preparation can be selected. In the case of protein-based biopharmaceuticals, powdering with excellent storage stability is often selected due to stability problems. It is desirable that the culture supernatant of the present invention is also produced as a solid preparation in order to improve stability and shelf life.

  In the agent of the present invention, a culture supernatant as an active ingredient may be prepared together with a pharmaceutically acceptable carrier or medium. Pharmaceutically acceptable carriers or vehicles include, for example, excipients, stabilizers, solubilizers, emulsifiers, suspending agents, buffers, isotonic agents, antioxidants, or preservatives. Are acceptable substances. Further, a polymer material such as polyethylene glycol (PEG) or a conjugation inhibitor such as cyclodextrin can also be used. Examples of excipients are those that themselves have no pharmacological action, such as starch or lactose. Examples of stabilizers are albumin, gelatin, sorbitol, mannitol, lactose, sucrose, trehalose, maltose, and glucose. Of these, sucrose or trehalose is preferred. Examples of solubilizers are ethanol, glycerin, propylene glycol, and polyethylene glycol. Examples of emulsifiers are lecithin, aluminum stearate or sorbitan sesquioleate. Examples of suspending agents are macrogol, polyvinylpyrrolidone (PVP), or carmellose (CMC). Examples of tonicity agents are sodium chloride and glucose. Examples of buffers are citrate, acetate, boric acid, and phosphate. As the aqueous medium for diluting the culture supernatant preparation, for example, an aqueous solution for injection in which the osmotic pressure and pH are adjusted to near blood values and the salt concentration and the like are adjusted may be used as appropriate. Ringer's solution such as acetated Ringer's solution, other infusions, physiological saline, glucose solution, and the like can be used, but are not limited thereto. Examples of antioxidants are ascorbic acid, sodium bisulfite, and sodium pyrosulfite. Examples of preservatives are phenol, thimerosal, and benzalkonium chloride.

  Agents containing the culture supernatant of the present invention include intravenous administration, intraarterial administration, intramuscular administration, subcutaneous administration, intraperitoneal administration, intranasal administration, spinal cord intraluminal transplantation, intraarticular transplantation, intragingival injection, application, etc. Can be administered using a known administration method. The agent of the present invention may be directly injected into an affected part or a target site, or the agent of the present invention can be administered by opening the affected part by surgical operation. Any optimal administration method is possible depending on the disease to be treated. When intravenous injection is selected as a transplantation method, it is preferable to administer the culture supernatant in a dose of 1 mL or more and 1,000 mL or less, more preferably 30 mL or more and 300 mL or less, as one administration unit. The dose may be appropriately adjusted by measuring the antibody value against an IgA antibody or an IgG antibody as shown in Examples described later.

  The present invention also provides a method for enhancing the antibody-producing ability, which comprises a step of administering a culture supernatant of mesenchymal stem cells to the subject in order to enhance the antibody-producing ability of the subject (human or non-human mammal). I do.

  The present invention also provides use of a culture supernatant of a mesenchymal stem cell for producing an antibody-producing ability enhancer.

[Production Example 1]
As described below, a culture supernatant of adipose tissue-derived mesenchymal stem cells was obtained using a serum-free medium (BMS medium) of Biomimetics Sympathies Co., Ltd.

  At a clinic or hospital with appropriate facilities, a doctor gave an explanation and subcutaneous fat was provided by a person who obtained informed consent and agreed to donate the tissue, or who obtained consent from a parent or guardian. This tissue was dispersed by enzymatic treatment and centrifuged to obtain fractions containing various cells.

  The obtained fraction was seeded on a plastic culture dish containing a BMS medium to obtain cells showing adhesiveness to the plastic. The obtained cells were examined for 1) plastic adhesion, 2) proliferation by culture, and 3) expression pattern of cell surface antigen marker, and confirmed that they were mesenchymal stem cells.

  The culture solution used for culturing the cells was collected, and the supernatant from which impurities were removed by centrifugation was used as the culture supernatant of the present invention.

[Production Example 2]
As described below, a culture supernatant of adipose tissue-derived mesenchymal stem cells was obtained using a serum-free medium (Lonza medium) from Lonza.

  Fractions containing various cells derived from adipose tissue obtained in the same manner as in Production Example 1 were seeded on a plastic culture dish containing Lonza medium, and cells showing adhesiveness to plastic were obtained. The obtained cells were examined for 1) plastic adhesion, 2) proliferation by culture, and 3) expression pattern of cell surface antigen marker, and confirmed that they were mesenchymal stem cells.

  The culture solution used for culturing the cells was collected, and the supernatant from which impurities were removed by centrifugation was used as the culture supernatant of the present invention.

[Production Example 3]
As described below, a culture supernatant of adipose tissue-derived mesenchymal stem cells was obtained using a serum-free medium (thermo medium) manufactured by Thermo Fisher Scientific Co., Ltd.

  Fractions containing various cells derived from adipose tissue obtained in the same manner as in Production Example 1 were seeded on a plastic culture dish containing a thermo-medium to obtain cells showing adhesiveness to plastic. The obtained cells were examined for 1) plastic adhesion, 2) proliferation by culture, and 3) expression pattern of cell surface antigen marker, and confirmed that they were mesenchymal stem cells.

  The culture solution used for culturing the cells was collected, and the supernatant from which impurities were removed by centrifugation was used as the culture supernatant of the present invention.

[Production Example 4]
A culture supernatant of umbilical cord-derived mesenchymal stem cells using a BMS medium was obtained as follows.

  At a clinic or hospital with appropriate equipment, a doctor gave an explanation, and a cord was provided by someone who obtained informed consent and agreed to donate the tissue. The umbilical cord was cut into pieces with sterilized scissors or the like, dispersed by enzyme treatment, and centrifuged to obtain fractions containing various cells.

  The obtained fraction was seeded on a plastic culture dish containing a BMS medium to obtain cells showing adhesiveness to the plastic. The obtained cells were examined for 1) plastic adhesion, 2) proliferation by culture, and 3) expression pattern of cell surface antigen marker, and confirmed that they were mesenchymal stem cells.

  The culture solution used for culturing the cells was collected, and the supernatant from which impurities were removed by centrifugation was used as the culture supernatant of the present invention.

[Production Example 5]
A culture supernatant of mesenchymal stem cells derived from dental pulp using a BMS medium was obtained as follows.

  At a clinic or hospital with appropriate facilities, a doctor gave an explanation, and a person with informed consent and the consent of a guardian was provided with missing milk teeth. The milk teeth were crushed with sterilized pliers or the like, and the pulp was taken out. It was dispersed by enzyme treatment and centrifuged to obtain fractions containing various cells.

  The obtained fraction was seeded on a plastic culture dish containing a BMS medium to obtain cells showing adhesiveness to the plastic. The obtained cells were examined for 1) plastic adhesion, 2) proliferation by culture, and 3) expression pattern of cell surface antigen marker, and confirmed that they were mesenchymal stem cells.

  The culture solution used for culturing the cells was collected, and the supernatant from which impurities were removed by centrifugation was used as the culture supernatant of the present invention.

  Example 1 shows the effect of the culture supernatant preparation of the present invention on anti-OVA antibody production.

  0.5 ml of the adipose tissue-derived mesenchymal stem cell culture supernatant preparation of the present invention was administered to the abdominal cavity of aged mice (18 to 22 months old). Three weeks after the administration, 1 mg of OVA was orally administered (oral immunization) once a week for three weeks together with 10 μg of CT as an adjuvant. One week after the final immunization, feces and serum were collected, and the antibody titers of secretory IgA antibodies and serum IgG antibodies contained in feces against OVA contained in each sample were measured by ELISA.

  FIG. 1 shows the results obtained by administering a culture supernatant obtained by culturing adipose tissue-derived mesenchymal stem cells in a BMS medium to aged mice and measuring the antibody titer against OVA. Group A shows aged mice (18-22 months old) without administration of the culture supernatant preparation of the present invention, and Group B shows aged mice (18-22 months old), Shows the culture supernatant formulation administered. The left graph shows the amount of IgA antibody secreted into the intestine, and the right graph shows the amount of serum IgG antibody by antibody titer. The aged mice had reduced antibody-producing ability, and no antibodies against OVA were detected. On the other hand, in the aged mice to which the culture supernatant preparation of the present invention was administered, a remarkable increase in antibody production was observed.

  FIG. 2 shows the results obtained by administering the culture supernatant obtained by culturing adipose tissue-derived mesenchymal stem cells in Lonza medium to aged mice and measuring the antibody titer against OVA. As in FIG. 1, no antibody was detected in aged mice to which the culture supernatant preparation was not administered, but a marked increase in antibody production was observed in the aged mice to which the culture supernatant preparation of the present invention was administered.

  FIG. 3 shows the results obtained by administering a culture supernatant obtained by culturing adipose tissue-derived mesenchymal stem cells in a thermo medium to an aged mouse and measuring the antibody titer against OVA. As in FIGS. 1 and 2, no antibody was detected in aged mice to which the culture supernatant was not administered, but a marked increase in antibody production was observed in aged mice to which the culture supernatant of the present invention was administered.

  As described above, regardless of the type of culture medium, the culture supernatant obtained from adipose tissue-derived mesenchymal stem cells cultured in a serum-free medium shows that intestinal secretory antibody (secretory IgA) and serum antibody ( It can be seen that the productivity of serum IgG) is significantly increased.

  Example 2 shows the effect of the culture supernatant preparation of the present invention on anti-CT antibody production.

  0.5 ml of the adipose tissue-derived mesenchymal stem cell culture supernatant preparation of the present invention was administered to the abdominal cavity of aged mice (18 to 22 months old). Three weeks after the administration, 10 μg of CT was orally administered (oral immunization) once a week for three weeks. One week after the final immunization, feces and serum were collected, and the antibody titers of secretory IgA antibody and serum IgG antibody to CT contained in each sample were measured by ELISA.

  FIG. 4 shows the results obtained by administering a culture supernatant obtained by culturing adipose tissue-derived mesenchymal stem cells in a BMS medium to an aged mouse and measuring the antibody titer against CT. Similar to the antibody production against OVA (Fig. 1), no antibody was detected in aged mice without administration of the culture supernatant, but a marked increase in antibody production was observed in aged mice administered with the culture supernatant of the present invention. Was seen.

  FIG. 5 shows the results of measuring the antibody titer to CT by administering a culture supernatant obtained by culturing adipose tissue-derived mesenchymal stem cells in Lonza medium to aged mice. As in FIG. 2, no antibody was detected in aged mice to which the culture supernatant preparation was not administered, but a marked increase in antibody production was observed in the aged mice to which the culture supernatant preparation of the present invention was administered.

  FIG. 6 shows the results obtained by administering a culture supernatant obtained by culturing adipose tissue-derived mesenchymal stem cells in a thermo medium to an aged mouse and measuring the antibody titer against CT. As in FIG. 3, no antibody was detected in aged mice without administration of the culture supernatant preparation, but a marked increase in antibody production was observed in aged mice administered with the culture supernatant preparation of the present invention.

  From the above, it can be seen that culture supernatants obtained from adipose tissue-derived mesenchymal stem cells cultured in various serum-free media significantly increase the ability to produce antibodies to various antigens.

  Example 3 shows the effect on antibody production of a culture supernatant preparation derived from another tissue.

  FIG. 7 shows the results obtained by administering a culture supernatant preparation obtained by culturing umbilical cord-derived mesenchymal stem cells in a BMS medium to aged mice in the same manner as in Example 1, and measuring the antibody titer against OVA. No antibody was detected in aged mice to which the culture supernatant preparation was not administered, but a marked increase in antibody production was observed in the aged mice to which the culture supernatant preparation of the present invention was administered.

  FIG. 8 shows the results obtained by administering a culture supernatant preparation obtained by culturing dental pulp-derived mesenchymal stem cells in a BMS medium to aged mice in the same manner as in Example 1 and measuring the antibody titer against OVA. As in the case of adipose tissue-derived (Fig. 1), no antibody was detected in aged mice without administration of the culture supernatant, but a marked increase in antibody production was observed in aged mice administered with the culture supernatant of the present invention. Was done.

  FIG. 9 shows the results obtained by administering a culture supernatant preparation obtained by culturing umbilical cord-derived mesenchymal stem cells in a BMS medium to aged mice in the same manner as in Example 2 and measuring the antibody titer against CT. No antibody was detected in aged mice to which the culture supernatant preparation was not administered, but a marked increase in antibody production was observed in the aged mice to which the culture supernatant preparation of the present invention was administered.

  FIG. 10 shows the results obtained by administering a culture supernatant obtained by culturing dental pulp-derived MSCs in a BMS medium to aged mice in the same manner as in Example 2 and measuring the antibody titer against CT. No antibody was detected in aged mice to which the culture supernatant preparation was not administered, but a marked increase in antibody production was observed in the aged mice to which the culture supernatant preparation of the present invention was administered.

  From the above, it is understood that culture supernatants obtained from mesenchymal stem cells derived from various tissues significantly increase the ability to produce antibodies against various antigens.

  As described above, antibody production ability was evaluated using culture supernatants of mesenchymal stem cells derived from various tissues, and it was shown that each of them had high antibody production enhancement ability.

The invention can be used in the pharmaceutical industry.

Claims (4)

  A culture supernatant preparation for enhancing antibody-producing ability, comprising a culture supernatant of mesenchymal stem cells as an active ingredient. The agent according to claim 1, wherein
An agent, wherein the mesenchymal stem cell is a human mesenchymal stem cell. The agent according to claim 1, wherein
The agent, wherein the antibody is an IgA antibody or an IgG antibody.   The agent according to claim 1, which is an infectious disease preventive agent, a vaccine enhancer, or an anticancer active agent.

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