WO2019240559A1 - Procédé d'amélioration de la viabilité cellulaire de cellules souches mésenchymateuses introduites par virus anti-cancéreux - Google Patents

Procédé d'amélioration de la viabilité cellulaire de cellules souches mésenchymateuses introduites par virus anti-cancéreux Download PDF

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WO2019240559A1
WO2019240559A1 PCT/KR2019/007276 KR2019007276W WO2019240559A1 WO 2019240559 A1 WO2019240559 A1 WO 2019240559A1 KR 2019007276 W KR2019007276 W KR 2019007276W WO 2019240559 A1 WO2019240559 A1 WO 2019240559A1
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stem cells
mesenchymal stem
cancer
virus
anticancer
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라정찬
김은영
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주식회사 알바이오
주식회사 네이처셀
라정찬
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    • C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
    • C12N5/06—Animal cells or tissues; Human cells or tissues
    • C12N5/0602—Vertebrate cells
    • C12N5/0652—Cells of skeletal and connective tissues; Mesenchyme
    • C12N5/0662—Stem cells
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
    • A61K35/12—Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells
    • A61K35/28—Bone marrow; Haematopoietic stem cells; Mesenchymal stem cells of any origin, e.g. adipose-derived stem cells
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
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    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K2039/51—Medicinal preparations containing antigens or antibodies comprising whole cells, viruses or DNA/RNA
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    • C12N2529/00—Culture process characterised by the use of electromagnetic stimulation
    • C12N2529/10—Stimulation by light
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    • C12N2760/00—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA ssRNA viruses negative-sense
    • C12N2760/00011—Details
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    • C12N2760/18411—Morbillivirus, e.g. Measles virus, canine distemper
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    • C12N2760/00—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA ssRNA viruses negative-sense
    • C12N2760/00011—Details
    • C12N2760/18011—Paramyxoviridae
    • C12N2760/18411—Morbillivirus, e.g. Measles virus, canine distemper
    • C12N2760/18451—Methods of production or purification of viral material

Definitions

  • the present invention relates to a method for producing mesenchymal stem cells containing an anticancer virus with improved cell viability and a cell therapy agent for treating cancer containing the stem cells prepared by the above method.
  • the present invention relates to the production of anticancer stem cell therapeutics having excellent activity and improved survival of mesenchymal stem cells by prolonging the replication time of anticancer viruses by introducing them into natural products and preventing the virus from lysing stem cells. .
  • Stem cells are cells that have the ability of self-replication and differentiate into two or more cells. Totipotent stem cells, pluripotent stem cells, and multipotent stem cells can be classified as a multipotent stem cell.
  • Pluripotent stem cells are pluripotent cells that can develop into a complete individual. Cells up to 8 cell stages after fertilization of eggs and sperm have these properties. If you transplant it into a single, complete entity.
  • Pluripotent stem cells are cells that can develop into a variety of cells and tissues derived from ectoderm, mesoderm, and endoderm.Inner cell mass located inside the blastocyst after 4-5 days of fertilization. They are called embryonic stem cells and differentiate into a variety of other tissue cells but do not form new life.
  • Multipotent stem cells are stem cells that can only differentiate into cells specific to the tissues and organs in which they are contained. In addition to growth and development, it is involved in maintaining homeostasis of adult tissues and inducing regeneration in tissue damage. Tissue-specific pluripotent cells are collectively called mesenchymal stem cells.
  • Mesenchymal stem cells (Rebecca SY Wong, et al., J Biomed Biotechnol 24: 2011, 2011) have been used for cell-based treatment in a variety of disease states, such as heart disease, osteoplastic insufficiency and spinal cord injury. It is becoming.
  • cancer is characterized by "uncontrolled cell growth,” and this abnormal cell growth forms a mass of cells called tumors that penetrate into surrounding tissues and, in severe cases, metastasize to other organs of the body. Sometimes. Academia is also called neoplasia.
  • Methods for treating cancer include surgery, radiation therapy, and chemotherapy for administering anticancer agents.
  • Bi-specific T-cell Engager (Bite), CAR-T (Chemeric antigen receptor T-cell or NK cells), anti-virus (Oncolytic Virus) from Immun checkpoint inhibitor
  • Numerous studies have been conducted for cancer conquest using various platforms such as.
  • the present inventors have a small amount of side effects, because the specific reaction to cancer only, and the anti-cancer efficacy of the anti-cancer virus is introduced as a result of the efforts to develop cancer treatments using stem cells, the anti-cancer virus introduced into the mesenchymal stem cells
  • the treatment with the following natural products prolongs the replication time of the anticancer virus and prevents the virus from lysing the stem cells, thereby improving the survival rate and survival of the stem cells and confirming that an anticancer stem cell therapy with excellent activity can be prepared.
  • the present invention has been completed.
  • An object of the present invention is to extend the replication time of oncolytic viruses through the treatment of natural substances in the mesenchymal stem cells introduced with oncolytic viruses, preventing the virus from lysing the stem cells, stem cells
  • the present invention comprises the steps of (a) preparing mesenchymal stem cells in a pellet; (b) infecting an anticancer virus to the mesenchymal stem cells in the pellet state; (c) centrifuging the mesenchymal stem cells infected with the anticancer virus; And (d) provides a method for producing mesenchymal stem cells containing an anti-cancer virus improved cell viability comprising the step of obtaining the mesenchymal stem cells into which the anti-cancer virus is introduced.
  • the present invention also provides a cell therapy agent for treating cancer containing mesenchymal stem cells prepared by the above method as an active ingredient.
  • the present invention also provides a method for treating cancer comprising administering to the subject a mesenchymal stem cell prepared by the above method.
  • the present invention also provides the use of mesenchymal stem cells prepared by the above method for use in the treatment of cancer.
  • the present invention also provides a cell therapy comprising mesenchymal stem cells prepared by the above method for use in the treatment of cancer.
  • the present invention also provides the use of mesenchymal stem cells prepared by the above method for the manufacture of a medicament for the treatment of cancer.
  • 1 is a fluorescence micrograph confirming MVeGFP by FITC fluorescence after measles virus infection in adipose derived stem cells.
  • Figure 2 confirms CD46, CD150, nectin-4 expression by FACS in cancer cell lines.
  • Figure 3 confirms the ability of MVeGFP killing in cancer cell lines and stem cells by the CPE (cytopathic effect) assay.
  • Figure 4 confirms the ability of MVeGFP killing in 1000TCID50 / ml breast cancer cell line by CPE assay.
  • Figure 6 confirms the measles virus infection in the flask and pellets of the CPE degree.
  • Figure 7 shows the CPE after varying the time and concentration of measles virus infection in pelleted cells.
  • Figure 8a confirms the survival rate of the virostem not cultured after measles virus infection.
  • Figure 8b confirms the survival rate of virostem cultured for 4 days after measles virus infection.
  • the anti-cancer virus is introduced into the mesenchymal stem cells, and then treated with natural products to prolong the replication time of the anti-cancer virus and prevent the virus from lysing the stem cells.
  • the anticancer stem cell therapy with excellent activity was prepared by improving the survival rate and survival time.
  • stem cells infected with the measles virus prepared by the method of the present invention was confirmed that the period of 80% survival was significantly increased.
  • the present invention is a point of consistency, (a) preparing mesenchymal stem cells in a pellet; (b) infecting an anticancer virus to the mesenchymal stem cells in the pellet state; (c) centrifuging the mesenchymal stem cells infected with the anticancer virus; And (d) relates to a method for producing mesenchymal stem cells containing an anti-cancer virus improved cell viability comprising the step of obtaining the mesenchymal stem cells into which the anti-cancer virus is introduced.
  • the mesenchymal stem cells of the step (a) is preferably cultured in a medium containing aspirin, the concentration of the aspirin is preferably 0.1mM to 1mM, but is not limited thereto.
  • the medium containing the aspirin preferably further comprises vitamin C, but is not limited thereto.
  • the medium is preferably DMEM or K-SFM containing 5-10% FBS and NAC (N-acetyl Cystein), and more preferably calcium, rEGF, insulin and hydrocortisone.
  • FBS N-acetyl Cystein
  • the present invention is not limited thereto.
  • the mesenchymal stem cells of the step (a) is preferably pretreated with vitamin C, but is not limited thereto.
  • mesenchymal stem cells with improved cancer cell proliferation inhibitory ability was prepared. That is, it may be a stem cell having anticancer function prepared by culturing fat-derived mesenchymal stem cells cultured by pretreatment with vitamin C in a medium containing aspirin, and fat-derived mesenchyme in a medium containing vitamin C and aspirin.
  • Stem cells may be prepared by culturing stem cells, and may also be anti-cancer function prepared by culturing fat-derived mesenchymal stem cells pre-treated with vitamin C in a medium containing vitamin C and aspirin.
  • Branches may be stem cells. We named all of these cells "Angel-Stem Cells" (WO / 2018/021879).
  • the degree of anti-cancer virus infection in the pellet state and the flask state of the stem cells was analyzed, and it was confirmed that the measles virus infection degree was excellent in the stem cell in the pellet state.
  • the mesenchymal stem cells of the step (b) is preferably 1 x 10 5 ⁇ 1 x 10 6 cells, more preferably 1 x 10 5 ⁇ 3 x 10 5 cells, most preferably Preferably 2 x 10 5 cells, but is not limited thereto.
  • the mesenchymal stem cells may be derived from a tissue selected from the group consisting of fat, uterus, bone marrow, muscle, placenta, umbilical cord blood, urine, hair follicles and skin.
  • stem cell used in the present invention refers to a cell having the ability of self-replicating and differentiating into two or more cells, and "adult stem cell” refers to each organ of the embryo during development. Refers to stem cells appearing in the stage of formation or adulthood.
  • meenchymal stem cell used in the present invention is an undifferentiated stem cell isolated from human or mammalian tissue, and may be derived from various tissues.
  • umbilical cord-derived mesenchymal stem cells umbilical cord blood-derived mesenchymal stem cells, bone marrow-derived mesenchymal stem cells, adipose-derived mesenchymal stem cells, muscle-derived mesenchymal stem cells, nerve-derived mesenchymal stem cells, skin-derived mesenchymal stem cells , Amnion derived mesenchymal stem cells and placental derived mesenchymal stem cells, and techniques for isolating stem cells from each tissue are already known in the art.
  • fat-derived stem cell used in the present invention is an undifferentiated stem cell isolated from adipose tissue, and the separation method may be as follows.
  • the suspension containing fat suspended in physiological saline obtained from liposuction and then treated with trypsin of the stem cell layer attached to the culture vessel such as a flask and recovered, or scraped with a scraper to directly suspended in a small amount of physiological saline
  • Adipose-derived mesenchymal stem cells can be separated by a method such as recovery.
  • adult tissue-derived adult stem cells or "fatty tissue-derived mesenchymal stem cells” are undifferentiated adult stem cells isolated from adipose tissue, and may be abbreviated herein as “fat stem cells”. This can be obtained through conventional methods known in the art.
  • a conventional medium known in the art to be suitable for culturing stem cells may be used.
  • DMEM Denssion Medium
  • Keratinocyte-SFM Keratinocyte serum free medium
  • IMSC Iscove's Modified Dulbecco's Medium
  • F12 Nutrient Mixture F-12
  • DMEM / F12 Dulbecco's Modified Eagle Medium: Nutrient Mixture F-12
  • Adipose stem cell culture medium may be supplemented with an additive that promotes proliferation of the undifferentiated phenotype of adipose stem cells while inhibiting differentiation.
  • the medium generally contains neutral buffers (such as phosphates and / or high concentrations of bicarbonate) and protein nutrients (such as serum, such as FBS, serum substitutes, albumin, or essential and non-essential amino acids, such as glutamine) in isotonic solutions. can do.
  • lipids fatty acids, cholesterol, HDL or LDL extracts of serum
  • other components found in most preservative media of this kind such as insulin or transferrin, nucleosides or nucleotides, pyruvate salts, any ionized form or salt
  • Sugar sources such as glucose, selenium, glucocorticoids such as hydrocortisone and / or reducing agents such as ⁇ -mercaptoethanol.
  • the medium also contains anti-clumping agents, such as those sold by Invitrogen (Cat # 0010057AE), with the aim of preventing the cells from adhering to each other, adhering to the vessel wall, or forming too large a bundle. It can be beneficial to do so.
  • anti-clumping agents such as those sold by Invitrogen (Cat # 0010057AE)
  • the medium for obtaining or culturing the adipose stem cells used in one embodiment of the present invention is a basal medium selected from the group consisting of DMEM, Defined Keratinocyte-SFM, ⁇ -MEM, IMDM, F12 and DMEM / F12, L It is preferable to contain aspirin in the medium composition for mesenchymal stem cell culture containing ascorbic acid 2-phosphate (vitamin C), fetal bovine serum and N-acetyl-L-cysteine.
  • vitamin C ascorbic acid 2-phosphate
  • fetal bovine serum fetal bovine serum
  • N-acetyl-L-cysteine N-acetyl-L-cysteine
  • the medium is 0.05-1 mM ascorbic acid 2-phosphate, 2-20% fetal bovine serum, 0.2-20 mM N-acetyl-L-cysteine and 0.1 to It may be characterized by containing 1mM aspirin, but is not limited thereto.
  • the anticancer virus is preferably a measles virus, more preferably MV (Edmonston strain), but is not limited thereto.
  • Measles virus is a malignant tumor (Msaouel P et al., Curr Pharm Biotechnol . 13 (9): 1732-41, 2012), cancer stem cell (CSC) (Fang Huang et al., World J Gastroenterol . 22 (35): 7999-8009, 2016), lung cancer (Zhao D et al., Oncol Rep . 29 (1): 199-204, 2013; Ong HT et al., J Hepatol . 59 (5): 999-1006, 2013), Blood tumors (D Grote et al., Blood . 97 (12): 3746-54, 2001), ovarian cancer (Zhou S et al., Cancer Lett .
  • CSC cancer stem cell
  • the anticancer virus of step (b) may be characterized in that the attenuated measles vaccine virus.
  • the attenuated virus may be a commercially available one, or may be used by attenuating and further attenuating a wild type virus or a commercially available attenuated virus.
  • MVeGFP a measles virus tagged with GFP
  • the anticancer virus of step (b) is preferably 1 x 10 5 ⁇ 1 x 10 7 TCID50, more preferably 1 x 10 5 ⁇ 5 x 10 6 TCID50, most preferably 1 x 10 6 TCID50, but is not limited thereto.
  • step (b) the infection of step (b) is preferably performed for 30 minutes to 2 hours at 36 ⁇ 37 °C, more preferably 30 minutes, but is not limited thereto.
  • the natural product is preferably an anticancer agent or an antioxidant, but is not limited thereto.
  • the anticancer agent is preferably aspirin or paclitaxel
  • the antioxidant is preferably alder extract or vitamin C, but is not limited thereto.
  • Treatment of these natural products with stem cells infected with measles virus or measles virus can increase the replication time of the virus, thereby prolonging the time for the virus to lyse the cells.
  • Stem cells infected with the measles virus of the present invention can be infected with stem cells by pretreatment of the measles vaccine virus itself with natural products to prevent the infected virus from lysing the cells, or the natural products can be treated to the stem cells infected with measles virus.
  • UV light irradiation may be further performed.
  • the time when the virus-infected mesenchymal stem cells survive more than 80% is only 48 hours (Mader EK et al., Clin Cancer Res . 1; 15; (23): 7246-55, 2009).
  • the time when the survival rate of mesenchymal stem cells is 80% or more is increased. Increased over 7 days.
  • the mesenchymal stem cells of the step (d) is preferably not further cultured after the anticancer virus is introduced, but is not limited thereto.
  • the present invention relates to a cell therapy agent for treating cancer containing mesenchymal stem cells prepared by the above method as an active ingredient.
  • the present invention relates to a method for treating cancer comprising administering to the subject a mesenchymal stem cell prepared by the above method.
  • the present invention relates to mesenchymal stem cells prepared by the above method for use in the treatment of cancer.
  • the present invention relates to a cell therapeutic agent comprising mesenchymal stem cells prepared by the above method for use in the treatment of cancer.
  • the present invention relates to mesenchymal stem cells prepared by the above method for the manufacture of a medicament for the treatment of cancer.
  • the anticancer stem cell therapy prepared by the method of the present invention can be administered in the form of a substantial product.
  • the anticancer stem cell therapeutic agent of the present invention can be produced without infecting the anticancer virus to the stem cells in the pellet state and then centrifuged, and further undergoing a culture step.
  • the cancer is lung cancer, blood tumor. Ovarian cancer, myeloma, breast cancer, brain cancer, rectal cancer, colon cancer, colorectal adenocarcinoma, osteosarcoma or cancer stem cells, but is not limited thereto.
  • the anticancer effect of the anticancer virus was excellent in breast cancer cells, and the effects were the same for both the estrogen-dependent breast cancer cell line (MCF-7) and the estrogen-independent breast cancer cell line (MDA-MB-231).
  • the mesenchymal stem cells may be derived from a tissue selected from the group consisting of fat, uterus, bone marrow, muscle, placenta, umbilical cord blood, urine, hair follicles and skin.
  • Cancer is characterized by uncontrolled cell growth, which results in the formation of cell masses called tumors that infiltrate surrounding tissues and, in severe cases, metastasize to other organs of the body.
  • anticancer includes not only the treatment of cancer diseases, ie, inhibiting the proliferation of cancer cells or cancer stem cells, or killing cancer cells or cancer stem cells, but also the prevention of cancer diseases, that is, increasing resistance to cancer before the onset of cancer. It is interpreted as. Therefore, the terms “cancer prevention or treatment” or “inhibition of cancer proliferation” and the term “anticancer” are used interchangeably herein.
  • cancer cells include cells with abnormal cell growth due to genetic modification during normal cell proliferation and growth, and cells with aggressive other organ mobility, which may be referred to as an ex.
  • cancer stem cells are known to exist in tumors and are thought to be caused by abnormal metastasis of genetic information of normal stem cells. Cancer stem cells are maintained and proliferated due to the presence of a microenvironment, a niche for their survival, and it is known that normal cells, immune-related cells, or differentiated cancer cells existing around them affect these properties.
  • cell therapeutic injection product or “cell therapeutic agent” refers to a parenteral administration containing stem cells for injection of a defect of a tissue, that is, injected into or near a defect in the form of an injection to correct a defect. It means a pharmaceutical composition that can be.
  • treating reverses, alleviates, inhibits, or prevents the disease or condition to which the term applies, or one or more symptoms of the disease or condition, Means that.
  • treatment refers to the act of treating when “treating” is defined as above.
  • treatment or “therapy” of cancer includes one or more of the following:
  • Cancer is an intractable chronic disease that, even if treated with surgery, radiation, and chemotherapy, in many cases does not heal fundamentally, suffers the patient, and ultimately leads to death. Surgery, chemotherapy and radiation therapy over the last few decades have not been the ultimate solution to cancer despite many advances.
  • the stem cells infected with the measles virus prepared by the method of the present invention is significantly increased to 7 days or more, showing a 80% survival rate, and can be administered in the form of a substantial stem cell therapeutic product.
  • the present inventors named the anticancer stem cell therapy combining the anticancer virus and the stem cells as "ViroSTEM.”
  • Human adipose tissue obtained from abdominal fat by liposuction was isolated and washed with PBS.
  • the tissue was chopped and digested for 2 hours at 37 ° C using DMEM medium containing collagenase type1 (1 mg / ml). After washing with PBS and centrifuged for 5 minutes at 1000rpm. The supernatant was suctioned and the pellet remaining on the bottom was washed with PBS, and then centrifuged at 1000 rpm for 5 minutes. Debris was removed by filtering on a 100 ⁇ m mesh, washed with PBS, and then cultured in DMEM medium containing 10% FBS, 2 mM NAC, and 0.2 mM ascorbic acid.
  • Adipose tissue-derived multipotent mesenchymal stem cells were isolated by subculture with alternating Keratinocyte-SFM medium every two days.
  • Adipose tissue-derived mesenchymal stem cells isolated as described above are stem cells cultured in a medium containing vitamin C, that is, vitamin C pre-treated mesenchymal stem cells.
  • Example 2 Cultivation of Mesenchymal Stem Cells in Aspirin-Containing Medium
  • Stem cells cultured in a medium containing vitamin C of Example 1 were inoculated in a 96-well cell culture plate at a concentration of 1 ⁇ 10 4 cells / plate, and then cultured overnight to attach and stabilize.
  • 0.5 mM of aspirin was added to Keratinocyte-SFM containing RKCM-N medium containing 5% FBS, 2 mM NAC, 0.2 mM ascorbic acid, 0.09 mM calcium, 5 ng / ml rEGF, 5 ug / ml insulin and 74 ng / ml Hydrocortisone. After exchange with a medium added at the concentration of and incubated for 24 hours.
  • Adipose tissue-derived mesenchymal stem cells obtained by culturing as described above were named Angel-Stem Cells (WO / 2018/021879) and were found to have excellent anticancer effects through direct and indirect co-culture with cancer cells. .
  • Example 1 Infection of adipose derived stem cells isolated in Example 1 or Example 2 with MVeGFP (GFP tagged measles virus) was confirmed. After purchasing Measles-GFP sold by Limanis, MVeGFP was amplified and titer was calculated to quantify MVeGFP, and the amplification method and titer identification method were established for the culture field of the present invention.
  • MVeGFP GFP tagged measles virus
  • the measles vaccine virus (10 6 TCID50) was infected by gently shaking every 30 minutes at 37 ° C in adipose derived stem cells (2X10 5 cells) in pellet or flask state. Stem cell infection of the measles vaccine virus obtained by culturing at 37 °C was confirmed by fluorescence microscopy (Fig. 1).
  • the killing ability of measles virus in various cancer cell lines can be measured by CD46, CD150 and nectin-4. Since the expression of three surface markers can be indirectly confirmed that the ability to kill due to measles virus is increased, the surface markers of each cancer cell line were confirmed by FACS assay (FIG. 2).
  • CPE cytopathic effect
  • Breast cancer cell lines were selected as cancer cell lines with high anti-cancer effect by infecting cancer cell lines with MVeGFP, and breast cancer cell lines (MCF-7: estrogen-dependent cell line and MDA-MB-231: estrogen-independent cell line) at 1000TCID50 / ml. Breast cancer killing ability of MVeGFP was confirmed (FIG. 4). As a result, when progressed to 1000TCID50 / ml, the positive control group showed CPE from day 2 and nearly 80% of CPE was formed on day 3, and both MCF-7 and MDA-MB-231 started to form CPE from day 3 It was.
  • MVeGFP infection in breast cancer cells was established by FACS assay to quantitatively determine the extent of infection (FIG. 5).
  • Measles virus was used after measles vaccine virus or after attenuating the measles virus virus once again.
  • the cells were infected in a flask or pellet state, and then the degree of CPE was confirmed.
  • the degree of infection was high in the cellular environment of the pellet state (FIG. 6), and the degree of infection was increased by giving a physical change by centrifugation after infection.
  • Example 6 Extending the survival time of stem cells infected with measles virus by natural product treatment
  • Virostem a mesenchymal stem cell incorporating measles virus, increased the survival by treating vitamin C or aspirin alone or in combination with antioxidants.
  • Example 7 Improvement of anticancer effect of measles virus infected stem cells by natural product treatment
  • the method for producing mesenchymal stem cells containing the anticancer virus according to the present invention extends the replication time of the anticancer virus through the treatment of natural substances and prevents the virus from dissolving the stem cells, thereby improving the viability and survival of the stem cells.
  • Stem cells containing the anticancer virus with improved activity can be prepared, and the anticancer stem cell therapy thus prepared has a markedly improved shelf life, which is very useful in medicine and industry.

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Abstract

La présente invention concerne un produit de thérapie cellulaire comprenant des cellules souches introduites par virus pour le traitement du cancer. Plus particulièrement, un virus anti-cancéreux est introduit dans des cellules souches mésenchymateuses, et est suivi d'un traitement avec une substance naturelle pour prolonger le temps de réplication du virus anti-cancéreux et empêcher le virus de lyser les cellules souches, ce qui permet de créer des cellules souches qui contiennent le virus anti-cancéreux présentant une excellente activité et d'améliorer la viabilité et la durée de vie des cellules souches mésenchymateuses. Le produit de cellule souche anti-cancéreux ainsi préparé présente une date d'expiration remarquablement améliorée et est très intéressant sur le plan industriel.
PCT/KR2019/007276 2018-06-15 2019-06-17 Procédé d'amélioration de la viabilité cellulaire de cellules souches mésenchymateuses introduites par virus anti-cancéreux WO2019240559A1 (fr)

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KR20140125943A (ko) * 2013-04-19 2014-10-30 창원대학교 산학협력단 지방유래줄기세포에 t항원을 도입한 adsc-t 세포의 배양액을 유효성분으로 포함하는, 염증질환의 예방 또는 치료용 조성물
KR101648915B1 (ko) * 2016-05-16 2016-08-17 주식회사 디지레이 줄기세포의 응집방지 방법 및 그 조성물
WO2018021879A1 (fr) * 2016-07-29 2018-02-01 라정찬 Procédé de production de cellules souches mésenchymateuses qui inhibent la prolifération de cellules cancéreuses

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KR20140125943A (ko) * 2013-04-19 2014-10-30 창원대학교 산학협력단 지방유래줄기세포에 t항원을 도입한 adsc-t 세포의 배양액을 유효성분으로 포함하는, 염증질환의 예방 또는 치료용 조성물
KR101648915B1 (ko) * 2016-05-16 2016-08-17 주식회사 디지레이 줄기세포의 응집방지 방법 및 그 조성물
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