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WO2020076015A1 - Composition destinée à induire la mort de cellules cancéreuses, comprenant un milieu conditionné préparé par culture de fibroblastes en condition hypoxique, et son procédé de préparation - Google Patents

Composition destinée à induire la mort de cellules cancéreuses, comprenant un milieu conditionné préparé par culture de fibroblastes en condition hypoxique, et son procédé de préparation Download PDF

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Publication number
WO2020076015A1
WO2020076015A1 PCT/KR2019/013063 KR2019013063W WO2020076015A1 WO 2020076015 A1 WO2020076015 A1 WO 2020076015A1 KR 2019013063 W KR2019013063 W KR 2019013063W WO 2020076015 A1 WO2020076015 A1 WO 2020076015A1
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cancer
medium
inducing
composition
cell death
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English (en)
Korean (ko)
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김동익
김애경
한규현
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Sungkyunkwan University
Samsung Life Public Welfare Foundation
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Sungkyunkwan University
Samsung Life Public Welfare Foundation
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    • 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
    • A61K35/33—Fibroblasts
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents

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  • the present invention is a composition for inducing cancer cell death, a medium composition for inducing cancer cell death or inducing inhibition of cancer cell proliferation, for treating or preventing cancer, comprising a conditioned medium prepared by culturing fibroblasts in a hypoxic environment It relates to a pharmaceutical composition and a method for preparing a composition for inducing cancer cell death.
  • Anticancer agents used as drug treatments are mainly synthesized by using monomolecular substances by organic or inorganic methods.
  • Traditional drug therapy in this way has many side effects, one of which is that the substance used as a drug is an artificially synthesized ex vivo derived substance, and the action point of an anti-cancer substance is to target an overexpressed protein.
  • Fibroblasts are cells, also called fibroblasts, and are cells unique to connective tissue that have ellipsoidal nuclei and spindle-like protoplasm characterized by good development of rough endoplasmic reticulum and Golgi. It also contains cells that fill the parenchymal cells in mesenchymal cells present in many internal organsviscera (collagen). It is known to have the ability to secrete the synthesis of interstitial substances such as proteoglycan, and to have a close relationship with myoplasts.
  • fibroblasts proliferate by serum or fibroblast growth factor (FGF), and when contact inhibition occurs, the shape deforms like a stone wall.
  • FGF fibroblast growth factor
  • fibroblasts when passage is repeated, it becomes 50s Almost known to die. It is used for screening of carcinoma by using the loss of function for contact inhibition of fibroblasts as described above, and studies of cell proliferation, especially growth factor. It is used a lot to study the signal transmission of.
  • the present inventors have shown the effect of inducing apoptosis specific to cancer cells, and while studying the naturally existing cells and proteins that have the potential to be used as anticancer agents, the fibroblast control medium prepared by culturing fibroblasts in a hypoxic culture environment While confirming that it is possible to induce apoptosis while inhibiting the proliferation of cancer cells, the present invention has been completed.
  • an object of the present invention is a composition for inducing cancer cell death, a medium composition for inducing cancer cell death or inducing cancer cell proliferation inhibition, comprising a conditioned medium prepared by culturing a fibroblast in a hypoxic environment. It is to provide a method for preparing a pharmaceutical composition for treatment or prevention and a composition for inducing cancer cell death.
  • the present invention provides a composition for inducing cancer cell death comprising a conditioned medium prepared by culturing fibroblasts in a hypoxic environment.
  • the present invention provides a method for inducing apoptosis of cancer cells, comprising treating the cancer cells with a conditioned medium prepared by culturing fibroblasts in a hypoxic environment.
  • the present invention provides a medium composition for inducing cancer cell death or inducing cancer cell proliferation inhibition, comprising a fibroblast culture medium cultured in a hypoxic environment.
  • the present invention provides a method for inducing cancer cell death or inhibiting cancer cell proliferation, comprising the step of treating a fibroblast culture medium cultured in a hypoxic environment with cancer cells.
  • the present invention provides a pharmaceutical composition for the treatment or prevention of cancer comprising the composition.
  • the present invention provides a method for preventing or treating cancer comprising administering the composition to an individual.
  • the present invention provides a method for preparing a composition for inducing cancer cell death, including; culturing a fibroblast in a hypoxic environment to prepare a conditioned medium.
  • the conditioned medium prepared by culturing fibroblasts in the hypoxic environment of the present invention When used, it is possible to induce apoptosis while suppressing the proliferation of cancer cells, and thus can be used as an anticancer agent.
  • the use of the conditioned medium prepared by the method of the present invention can specifically kill only cancer cells without inducing the death of surrounding normal cells, and thus can be effectively used as a clinically applicable cancer treatment or prevention pharmaceutical composition. You can.
  • FIG. 1 is a diagram showing the experimental results confirming the ability to inhibit Hela cell proliferation that occurs when fibroblast control media cultured in hypoxic conditions for 48 hours (2 days) and 72 hours (3 days) (* P ⁇ 0.05 , ** P ⁇ 0.01).
  • Figure 2A is a diagram showing the results of the degree of apoptosis (apoptosis) of Hela cells through an annexin V analysis by medium culture medium, serum-free medium conditions, 21% normal oxygen conditions, 1% hypoxic conditions medium.
  • Figure 2B is a graph showing the degree of cell death (apoptosis) of Hela cells through Annexin V analysis by medium culture medium, serum-free medium condition, 21% normal oxygen condition, 1% hypoxic condition medium (Figure 2B) Is (* P ⁇ 0.05, ** P ⁇ 0.01).
  • Figure 3 is a diagram showing the results of measuring the Caspase3 / 7 activity of cancer cells by medium culture medium, serum-free medium condition, 21% normal oxygen condition, 1% hypoxic condition medium (* P ⁇ 0.05, ** P ⁇ 0.01) .
  • Figure 4 is a diagram showing the experimental results of measuring the mitochondrial membrane potential change of cancer cells by medium culture medium, serum-free medium condition, 21% normal oxygen condition, 1% hypoxic condition medium (* P ⁇ 0.05, ** P ⁇ 0.01 ).
  • Figure 5A is a diagram showing the results of counting cancer cell cell cycles per medium culture medium, serum-free medium condition, 21% normal oxygen condition, and 1% hypoxic condition medium.
  • 5B is a diagram showing the results of displaying cancer cells corresponding to each cell cycle of cancer cells according to the basic culture medium, serum-free medium conditions, 21% normal oxygen conditions, and 1% hypoxic conditions medium (* P ⁇ 0.05, ** P ⁇ 0.01).
  • FIG. 6 is a diagram showing the experimental results confirming the cancer cell killing effect confirmed in the control medium prepared by culturing fibroblasts with hypoxic concentration conditions of 1% and 5% (* P ⁇ 0.05, ** P ⁇ 0.01).
  • the present invention provides a composition for inducing cancer cell death, comprising a conditioned medium prepared by culturing fibroblasts in a hypoxic environment.
  • the present invention provides a method for inducing apoptosis of cancer cells, comprising treating the cancer cells with a conditioned medium prepared by culturing fibroblasts in a hypoxic environment.
  • fibroblast used in the present invention is a cell called fibroblast, which is also called a fibroblast, and is an indigenous cell of connective tissue, which is an elliptical nucleus and spindle characterized by good development of the rough endoplasmic reticulum and the Golgi apparatus. Refers to the cell with the mercury protoplasm.
  • the term 'culture media' refers to a culture medium capable of supporting the growth and survival of cells under in vitro culture conditions, and includes all mediums used in the art suitable for culturing fibroblasts. Includes.
  • medium and culture conditions can be selected according to the type of cells.
  • the medium used for the culture is preferably a cell culture minimum medium (CCMM), and generally contains a carbon source, a nitrogen source, and a trace element component.
  • CCMM cell culture minimum medium
  • Such cell culture media include, for example, Dulbecco's Modified Eagle's Medium (DMEM), Minimal Essential Medium (MEM), Basal Medium Eagle (BME), RPMI1640, F-10, F-12, ⁇ MEM ( ⁇ Minimal essential Medium), GMEM ( Glasgow's Minimal essential Medium), Iscove's Modified Dulbecco's Medium, etc., but are not limited to these.
  • DMEM Dulbecco's Modified Eagle's Medium
  • MEM Minimal Essential Medium
  • BME Basal Medium Eagle
  • RPMI1640 F-10, F-12
  • ⁇ MEM ⁇ Minimal essential Medium
  • GMEM Glasgow's Minimal essential Medium
  • Iscove's Modified Dulbecco's Medium etc., but are not limited to these.
  • conditioned medium used in the present invention means a culture supernatant obtained by culturing fibroblasts, or a fibroblast from which fibroblasts are removed, and "fibroblast culture supernatant", It can be used interchangeably with “fibroblast culture medium” or "fibroblast culture medium”.
  • the medium of the present invention may further include a nutrient mixture (Nutrient Mixture).
  • the nutritional mixture is a mixture containing various amino acids, vitamins, and inorganic salts commonly used in cell culture, and may be prepared by mixing the amino acid, vitamins, and inorganic salts, or a commercially produced nutritional mixture.
  • Commercially prepared nutrient mixtures include, for example, M199, MCDBllO, MCDB202, MCDB302, but are not limited thereto.
  • fibroblasts were cultured in a hypoxic environment to prepare a conditioned medium, and it was confirmed that the prepared conditioned medium exhibited anticancer activity.
  • the anti-cancer activity identified in one embodiment of the present invention is one or more activities selected from the group consisting of inhibiting cancer cell proliferation, inducing cancer cell death (apoptosis), and arresting cancer cell cycles.
  • apoptosis-inducing activity is due to an increase in caspase 3/7 activity or a decrease in mitochondrial membrane potential.
  • the “hypoxic” environmental condition refers to an oxygen concentration lower than the normal atmospheric oxygen concentration of 21%, and may be 0.5% to 7% oxygen concentration, preferably 0.7% to 5% oxygen concentration, , More preferably, it may be a culture condition containing an oxygen concentration of 1%.
  • the present invention provides a medium composition for inducing cancer cell death or inhibiting cancer cell proliferation, comprising a fibroblast culture medium cultured in a hypoxic environment.
  • the present invention provides a method for inducing cancer cell death or inhibiting cancer cell proliferation, comprising the step of treating a fibroblast culture medium cultured in a hypoxic environment with cancer cells.
  • the medium composition comprising the fibroblast culture medium cultured in the hypoxic environment of the present invention can induce the death of cancer cells by increasing the caspase 3/7 activity of the cancer cells treated with the composition or reducing mitochondrial membrane potential, It can be effectively used to induce cancer cell death.
  • the medium composition comprising the fibroblast culture medium cultured in the hypoxic environment of the present invention can detain the cell cycle of the treated cancer cells, and thus can be effectively used to suppress cancer cell proliferation.
  • the present invention provides a pharmaceutical composition for preventing or treating cancer comprising the composition.
  • the present invention provides a method for preventing or treating cancer comprising administering the composition to an individual.
  • the term 'cancer' in the present invention refers to a group of diseases having a characteristic that cells can overproliferate and infiltrate into surrounding tissues when the normal apoptotic balance is broken.
  • the cancer of the present invention is brain tumor, benign astrocytoma, malignant astrocytoma, pituitary adenoma, meningioma, cerebral lymphoma, oligodendroma, intracranial endothelial, epithelial cell tumor, brain stem tumor, head and neck tumor, larynx cancer, oropharyngeal cancer, nasal / sinus cancer, nasopharyngeal cancer , Salivary gland cancer, hypopharyngeal cancer, thyroid cancer, thoracic tumor, small cell lung cancer, non-small cell lung cancer, thymic cancer, mediastinal tumor, esophageal cancer, breast cancer, male breast cancer, abdominal tumor, stomach cancer, liver cancer, gallbladder cancer, biliary cancer, pancreatic cancer
  • the pharmaceutical composition of the present invention in addition to a composition for inducing cancer cell death, including a culture medium prepared by culturing fibroblasts in a hypoxic environment, is a pharmaceutically acceptable carrier, that is, saline, sterile water, Ringer's solution, buffered saline, cyclodextrin, and dext. Rose solution, maltodextrin solution, glycerol, ethanol, liposomes and one or more of these components may be used in combination, and if necessary, may further include other conventional additives such as antioxidants and buffers.
  • a pharmaceutically acceptable carrier that is, saline, sterile water, Ringer's solution, buffered saline, cyclodextrin, and dext.
  • Rose solution, maltodextrin solution, glycerol, ethanol, liposomes and one or more of these components may be used in combination, and if necessary, may further include other conventional additives such as antioxidants and buffer
  • diluents, dispersants, surfactants, binders and / or lubricants can be additionally added to formulate into injectable formulations such as aqueous solutions, suspensions, emulsions, pills, capsules, granules or tablets.
  • injectable formulations such as aqueous solutions, suspensions, emulsions, pills, capsules, granules or tablets.
  • it can be preferably formulated according to each component using an appropriate method in the art.
  • the pharmaceutical composition of the present invention is not particularly limited in the formulation, but is preferably formulated as an injection or inhalation.
  • the method of administration of the pharmaceutical composition of the present invention is not particularly limited, but may be administered parenterally or orally, such as intravenous, subcutaneous, intraperitoneal, inhalation or topical application depending on the desired method.
  • the dosage range varies according to the patient's weight, age, sex, health status, diet, administration time, administration method, excretion rate, and disease severity.
  • the daily dosage refers to the amount of the therapeutic substance of the present invention sufficient for treatment for a reduced disease state by being administered to an individual in need of treatment.
  • the effective amount of a therapeutic agent depends on the particular compound, disease condition and severity thereof, and the individual in need of treatment, which can be routinely determined by those skilled in the art.
  • the dosage of the composition according to the present invention to the human body may vary depending on the patient's age, body weight, sex, dosage form, health condition, and degree of disease, and may be based on an adult patient having a body weight of 70 kg. In this case, it is generally 0.0001 to 1000 mg / day, preferably 1 to 500 mg / day, and may be dividedly administered once to several times a day at regular time intervals.
  • 'individual' refers to a subject in need of treatment of cancer, and more specifically, a human or non-human primate, a mouse, a rat, a dog, a cat, a horse, and a cow.
  • a human or non-human primate a mouse, a rat, a dog, a cat, a horse, and a cow.
  • the present invention provides a method for preparing a composition for inducing cancer cell death, including; culturing a fibroblast in a hypoxic environment to prepare a conditioned medium.
  • the step of culturing the fibroblasts of the present invention in a hypoxic environment to prepare a conditioned medium includes culturing the fibroblasts at an oxygen concentration lower than the normal atmospheric oxygen concentration of 21%, wherein the hypoxic environment is 0.5% to 7% oxygen It may be a concentration environment, but preferably may be a 0.7% to 5% oxygen concentration environment, and more preferably refers to an oxygen concentration environment of 1%.
  • the medium used in the step of preparing the conditioned medium of the present invention is, for example, DMEM (Dulbecco's Modified Eagle's Medium), MEM (Minimal Essential Medium), BME (Basal Medium Eagle), RPMI1640, F-10, F-12, ⁇ MEM ( ⁇ Minimal essential Medium), GMEM (Glasgow's Minimal essential Medium), Iscove's Modified Dulbecco's Medium, etc. can be used, but are not limited to these.
  • composition for inducing cancer cell death through the production method of the present invention When the composition for inducing cancer cell death through the production method of the present invention is used, inhibition of cancer cell proliferation and cell cycle arrest of cancer cells can be effectively induced to kill the cancer cells treated with the composition.
  • the fibroblasts used in the experiment were cultured for 2 weeks or more under normal atmospheric oxygen concentration conditions (21% O 2 ) and hypoxic concentration conditions (1% and 5% O 2 ), and then fibroblasts cultured in each oxygen condition environment 2x10 5 Dog cells in DMEM medium (Dulbeco's Modified Eagle's Media) (Gibco, Grand NY, USA) with 10% FBS (Fetal Bovine Serum) (Gibco, Grand NY, USA) and 1% antibiotic (penicillin / streptomycin) (Gibco, Grand NY, USA) was added to a 100 mm culture plate to which 10 ml of the culture medium solution was added, and cultured for 5 days.
  • DMEM medium Dulbeco's Modified Eagle's Media
  • FBS Fetal Bovine Serum
  • antibiotic penicillin / streptomycin
  • the cells were washed with 1 X PBS. Thereafter, 6 ml of DMEM without serum was added to the fibroblasts. After 24 hours, a medium was obtained, and the obtained medium was centrifuged at 300 g for 5 minutes. The supernatant was obtained and stored frozen at -80 ° C. Thereafter, in the following examples, the conditioned medium obtained in this example was used.
  • fibroblast control medium cultured under hypoxic conditions were added to each well of a 96-well white plate and cultured for 1 day. Thereafter, 100 ⁇ l of the fibroblast control medium obtained in Example 1 cultured under normal atmospheric oxygen concentration conditions (21% O 2 ) and low oxygen concentration conditions (1% O 2 ) was added to the medium. Thereafter, after incubation for 48 hours (2 days) and 72 hours (3 days), 100 ⁇ l of CellTiter-Glo assay 2.0 reagent (Promega, USA) was added and reacted for 10 minutes.
  • the basic culture medium means a basic culture medium to which 1% antibiotic, DMEM and 10% FBS are added
  • the serum-free medium condition adjustment medium refers to a culture medium of serum-free condition to which only 1% antibiotic and DMEM are added
  • the normal oxygen condition adjusting medium refers to a control medium obtained by culturing fibroblasts under normal oxygen conditions obtained in Example 1
  • the 1% hypoxic condition adjusting medium is used to control fibroblasts under 1% hypoxic conditions obtained in Example 1. It means the conditioned medium obtained by culturing.
  • fibroblast control medium prepared by culturing under hypoxic conditions can kill cancer cells.
  • Annexin V expression analysis was performed. 1.5x10 5 Hela cells were added to each well of a 6-well culture plate, and cultured for 1 day, obtained in Example 1 cultured under normal atmospheric oxygen concentration conditions (21% O 2 ) and low oxygen concentration conditions (1% O 2 ). 2 ml of fibroblast conditioned medium was added to the plate.
  • the Hela cells were cultured for 48 hours, and stained with FITC Annexin V apoptosis Detection Kit I (BD Pharmingen, SanDiego, CA, USA), followed by a FACS Caliber flow cytometer ( Results were analyzed using Becton-Dickinson, San Jose, CA, USA), and the results are shown in 2A and 2B, respectively.
  • the basic culture medium means a basic culture medium to which 1% antibiotic, DMEM and 10% FBS are added
  • the serum-free medium condition adjustment medium refers to a culture medium of serum-free condition to which only 1% antibiotic and DMEM are added
  • the normal oxygen condition adjusting medium refers to a control medium obtained by culturing fibroblasts under normal oxygen conditions obtained in Example 1
  • the 1% hypoxic condition adjusting medium is used to control fibroblasts under 1% hypoxic conditions obtained in Example 1. It means the conditioned medium obtained by culturing.
  • the basic culture medium means a basic culture medium to which 1% antibiotic, DMEM and 10% FBS are added
  • the serum-free medium condition adjustment medium refers to a culture medium of serum-free condition to which only 1% antibiotic and DMEM are added
  • the normal oxygen condition adjusting medium refers to a control medium obtained by culturing fibroblasts under normal oxygen conditions obtained in Example 1
  • the 1% hypoxic condition adjusting medium is used to control fibroblasts under 1% hypoxic conditions obtained in Example 1. It means the conditioned medium obtained by culturing.
  • the apoptosis marker caspase 3/7 activity increased in 1% hypoxic condition conditioned medium compared to the control serum-free medium conditioned medium. After 24 hours, it was confirmed that the activity was significantly increased. In particular, when 48 hours have elapsed, it was confirmed that the caspase 3/7 activity was increased by 2 times or more in the 1% hypoxic condition-adjusted medium as compared to the control-free serum-free medium condition.
  • the basic culture medium means a basic culture medium to which 1% antibiotic, DMEM and 10% FBS are added
  • the serum-free medium condition adjustment medium refers to a culture medium of serum-free condition to which only 1% antibiotic and DMEM are added
  • the normal oxygen condition adjusting medium refers to a control medium obtained by culturing fibroblasts under normal oxygen conditions obtained in Example 1
  • the 1% hypoxic condition adjusting medium is used to control fibroblasts under 1% hypoxic conditions obtained in Example 1. It means the conditioned medium obtained by culturing.
  • cancer cells were cultured in a control medium containing 21% normal oxygen condition and a culture medium containing 1% hypoxic condition compared to the control-free medium. It was confirmed that the mitochondrial membrane potential was significantly decreased. In particular, after 48 hours, it was confirmed that the membrane potential of cancer cells was significantly reduced when cultured in a 1% hypoxic condition-adjusted medium than a 21% normal-oxygen condition-adjusted medium.
  • the conditioned medium cultured in a hypoxic environment induced the death of cancer cells, thereby confirming a marked decrease in mitochondrial membrane potential.
  • the basic culture medium means a basic culture medium to which 1% antibiotic, DMEM and 10% FBS are added
  • the serum-free medium condition adjustment medium refers to a culture medium of serum-free condition to which only 1% antibiotic and DMEM are added
  • the normal oxygen condition adjusting medium refers to a control medium obtained by culturing fibroblasts under normal oxygen conditions obtained in Example 1
  • the 1% hypoxic condition adjusting medium is used to control fibroblasts under 1% hypoxic conditions obtained in Example 1. It means the conditioned medium obtained by culturing.
  • the M phase in which the cells divide during the cell cycle of the cultured cancer cells decreases and G0 / G1 phase without cell division And as the S phase is increased, it was confirmed that the effect of suppressing the overall proliferation of cancer cells due to the cell cycle deterrence effect can be exhibited comprehensively.
  • Example 7 Confirmation of hypoxic conditions under which an optimized cancer cell proliferation inhibitory effect of the fibroblast control medium is exhibited
  • fibroblast control medium cultured in the hypoxic conditions of Examples 2 to 6 an experiment was performed to confirm whether the cancer cell killing effect is the best when the hypoxic condition is to a certain extent.
  • Hela cells 1x10 4 cervical cancer cells, were added to each well of a 96-well white plate and cultured for 1 day. Thereafter, 100 ⁇ l of the fibroblast control medium obtained in Example 1 cultured under normal atmospheric oxygen concentration conditions (21% O 2 ) and low oxygen concentration conditions (1% O 2 ) was added to the medium. After incubation for 48 hours, hela cells were reacted for 10 minutes by adding 100 ⁇ l of CellTiter-Glo assay 2.0 reagent (Promega, USA).
  • the basic culture medium means a basic culture medium to which 1% antibiotic, DMEM and 10% FBS are added
  • the serum-free medium condition adjustment medium refers to a culture medium of serum-free condition to which only 1% antibiotic and DMEM are added
  • the normal oxygen condition adjusting medium refers to a control medium obtained by culturing fibroblasts under normal oxygen conditions obtained in Example 1
  • the 1% hypoxic condition adjusting medium is used to control fibroblasts under 1% hypoxic conditions obtained in Example 1.
  • Refers to a conditioned medium obtained by culturing, and a 5% hypoxic condition conditioned medium refers to a conditioned medium obtained by culturing fibroblasts under 5% hypoxic conditions obtained in Example 1.
  • the condition of the hypoxic concentration in which the anticancer effect is optimal is 1%.

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Abstract

La présente invention concerne une composition destinée à induire la mort de cellules cancéreuses, comprenant un milieu conditionné préparé par culture de fibroblastes en condition hypoxique, une composition de milieu destinée à induire la mort de cellules cancéreuses ou à induire l'inhibition de la prolifération de cellules cancéreuses, une composition pharmaceutique destinée à traiter ou à prévenir le cancer, et un procédé de préparation d'une composition pour l'induction de la mort de cellules cancéreuses. Le milieu conditionné préparé par culture de fibroblastes en condition hypoxique selon la présente invention peut être utilisé pour inhiber la prolifération de cellules cancéreuses et pour induire la mort de cellules cancéreuses, ce qui permet de trouver des applications dans des agents anticancéreux. De plus, le milieu conditionné préparé par le procédé de la présente invention peut être utilisé pour tuer spécifiquement les cellules cancéreuses uniquement, sans induire la mort des cellules normales voisines et, ainsi, peut être avantageusement utilisé dans une composition pharmaceutique, cliniquement applicable immédiatement, pour le traitement ou la prévention du cancer.
PCT/KR2019/013063 2018-10-10 2019-10-04 Composition destinée à induire la mort de cellules cancéreuses, comprenant un milieu conditionné préparé par culture de fibroblastes en condition hypoxique, et son procédé de préparation Ceased WO2020076015A1 (fr)

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US20040005704A1 (en) * 1998-11-18 2004-01-08 California Institute Of Technology Low oxygen culturing of central nervous system progenitor cells
US20110064786A1 (en) * 2008-03-07 2011-03-17 Page Raymond L Novel use of basic fibroblast growth factor in the de-differentiation of animal connective tissue cells
KR101610985B1 (ko) * 2008-11-14 2016-04-08 히스토젠, 인코포레이티드 암 치료를 위한 세포외 기질 조성물
KR20120089235A (ko) * 2009-07-10 2012-08-09 히스토젠, 인코포레이티드 저산소 조건 하에 배양된 세포로부터의 조정 배지 및 세포외 기질 조성물
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