Treatment of opioid addiction using fibroblasts and products thereof

Abstract

To provide methods and compositions related to the treatment or prevention of any type of addiction or substance abuse.SOLUTION: In some embodiments, methods comprise treating a patient addicted to opioids by administering a fibroblast population at a concentration sufficient for suppression of addiction-associated brain damage. In some embodiments, the fibroblasts express CD31 and / or CD73 markers. In some embodiments, fibroblasts are used to endow neuronal regeneration in order to overcome changes in the brain associated with addiction. Some embodiments relate to the stimulation of hippocampal regeneration subsequent to addiction induced damage.SELECTED DRAWING: None

Description

[Technical field]

[0001] This application was filed on March 19, 2019, and is incorporated herein by reference in its entirety. This application claims priority to U.S. Provisional Patent Application No. 62 / 820,721.

[0002] (Technical field) The present disclosure relates generally to cell biology and medicine embodiments. In certain embodiments, the present disclosure The present disclosure relates to regenerative medicine for the treatment of addiction. More particularly, the present disclosure relates to the treatment of opioid addiction. The present invention relates to compositions and methods comprising fibroblasts or products derived therefrom for treatment. [Background technology]

[0003] For example, dependence on opioids, either in the form of heroin or prescription drugs, is a serious health concern. In the United States, 568,699 people died from drug overdoses between 1999 and 2015. It is known that drug overdose deaths in the United States increased 1.2% from 2014 to 2015. This increased by 11.4% from 2010 to 2015, resulting in 52,404 deaths, of which This included 33,091 cases (63.1%) involving opioids. The largest increase over the past five years was in deaths involving synthetic opioids other than methadone (72.2%). %. There was demographic and geographic variation in overdose deaths involving various drugs. Therefore, the U.S. Centers for Disease Control and Prevention (CDC) has established a system for the management of all opioids, opioid subcategories, and - (i.e. prescription opioids, heroin, synthetic opioids), cocaine, potential for abuse Age-standardized death rates from overdoses involving certain psychostimulants (psychostimulants) compared with population statistics, urbanization, and The study looked at 31 states and the District of Columbia (DC). In 2016, 63,632 There were 10 drug overdose deaths, of which 42,249 (66.4%) were from opioids. From 2015 to 2016, deaths occurred across all drug categories examined. The largest overall increases were in cocaine (52.4%) and synthetic opioids (10 0%) and was caused by illicitly manufactured fentanyl (IMF). The increase is likely driven by demographics, urbanization levels, and state and DC The authors conclude that the opioid overdose epidemic in the United States continues to worsen. It is generally accepted that the conclusion that

[0004] Methadone maintenance treatment for opioid addiction reduces morbidity, mortality, and the prevalence of infectious diseases. However, it is limited to licensed specialist clinics in the United States, requires frequent clinic visits, and is difficult to treat with overdose. There is a high risk of administration.[5]

[0005] These issues have led to an increase in the use of buprenorphine to treat opioid addiction. There are numerous studies supporting the effectiveness of sublingual buprenorphine. Buprenorphine can be prescribed by office-based physician practices. There are some concerns about diversion and non-medical use of buprenorphine. Sublingual prenorphine is associated with poor adherence to treatment, cravings, withdrawal symptoms, and relapses. There are also concerns that this could increase the likelihood of cancer development [6, 7].

[0006] Generally speaking, addiction is a condition that requires the use of drugs and / or alcohol and inhibits consumption. It is defined as a chronic illness in which the patient is unable to control their intake of medication and has negative emotional feelings during the withdrawal period. Addiction not only affects the lives of addicts, but also places a huge burden on society and the economy. Sexual drugs exert profound anatomical and physiological effects on brain centers, affecting psychological, behavioral, and sensory - It has been shown that addictive substances cause motor changes, especially in the nucleus accumbens, amygdala, and abdominal cavity. Dopaminergic pathways in the lateral pallidum and hippocampus link the ventral tegmental area and prefrontal cortex via the limbic system It has been demonstrated that it affects the pathway [8].

[0007] There is now a safe therapeutic composition that helps to adequately treat and / or manage opioid addiction. There is a need for methods and regimens. Summary of the Invention

[0008] Embodiments of the present disclosure include methods and methods relating to the treatment or prevention of any type of addiction or substance abuse. In certain embodiments, the present disclosure includes methods and compositions for inhibiting addiction, such as opioid addiction. and / or the administration of fibroblasts and / or products derived therefrom for use in treating Current embodiments include, inter alia, compositions and methods relating to the treatment of addiction and substance abuse. In some embodiments, the compositions and methods involve the administration of fibroblasts and / or products derived therefrom. In some embodiments, the present invention relates to the treatment or inhibition of addiction or substance abuse by administering Blast cells express CD31 and / or CD73 with or without other markers In some embodiments, the drug is administered to a substance addict to reduce addiction and / or associated brain damage. In some embodiments, the fibroblast population is administered at a concentration sufficient to is used to induce neuronal regeneration to overcome addiction-related brain changes In some embodiments, the methods relate to stimulating hippocampal regeneration following addiction-induced injury. The disclosure also provides the use of fibroblasts as a cellular therapy for the amelioration of addictive conditions.

[0009] Here, embodiments include those for treating opioid addiction, for treating opioid use disorder, To treat opioid withdrawal symptoms, to reverse addiction, to prevent opioid addiction, To inhibit, to inhibit, to treat or ameliorate opioid addiction , to prevent, treat, or ameliorate opioid withdrawal symptoms, to assess the severity or duration of opioid addiction, To reduce the duration of opioid addiction, to prevent relapse, and to reduce the possibility of opioid tolerance. to reduce sexual activity, to reduce physical dependence, to prevent opioid abuse, or To overcome addiction-related hippocampal damage, fibroblasts were administered to induce additional associated changes in hippocampal function. and the use of fibroblasts for enhanced efficacy in treating opioid addiction. To select, to treat any addiction, to provide neuronal regeneration, to stimulate hippocampal regeneration stimulates the regeneration of neuronal precursor cells, inhibits the regeneration of neuronal precursor cells, and to enhance neuroprotection, inhibit excitotoxicity, prevent apoptosis of brain cells, or and compositions and formulations comprising fibroblasts for any combination thereof.

[0010] Certain aspects of the disclosure include the treatment of opioid addiction, the treatment of opioid withdrawal symptoms, the treatment of addictive states, Use of fibroblasts for the treatment, prevention or amelioration of opioid addiction, and opioid withdrawal symptoms suppression, reduction in severity or duration of opioid addiction, reduction in opioid tolerance, opioid For preventing abuse, overcoming hippocampal damage associated with addiction, or correcting changes associated with the addition of hippocampal function Selection of fibroblasts, use of fibroblasts for the treatment of any addiction, stimulation of hippocampal regeneration, neuroprotection Enhancement of progenitor cell renewal, inhibition of excitotoxicity, prevention of apoptosis of brain cells, or a combination of these. Regarding the match.

[0011] These method embodiments are disclosed throughout the disclosure. The embodiment may be implemented in the context of another embodiment discussed herein. Any of the methods described herein may exclude one or more steps described herein. In some embodiments, any one of the methods disclosed herein comprises the steps of: diagnosing , treating, ameliorating, inhibiting, and enriching.

[0012] A further aspect of the present disclosure relates to a method of treating an addiction in a subject, the method comprising: The method includes administering to the subject a composition comprising a fibroblast, a plurality thereof, or a product derived therefrom. wherein the cells express the CD31 and / or CD73 markers. In one embodiment, the cells include a) obtaining fibroblasts, and b) culturing said fibroblasts. c) obtaining a single cell suspension; and d) detecting the markers CD31 and CD7. and extracting from said single cell suspension fibroblasts expressing 3. In some embodiments, the fibroblast or cells thereof are generated by expressing CD45 and / or CD45+. or further expresses the CD34 marker.

[0013] In some embodiments, the derived product comprises supernatant from a fibroblast culture. In some embodiments, the supernatant from the fibroblast culture comprises at least one trophic factor. Trophic factors include hormones, cytokines, extracellular matrix, proteins, vesicles, antibodies, Or it may be granular.

[0014] In some embodiments, the methods of the present disclosure involve the expression of the markers CD31 and / or CD73. The fibroblasts can be enhanced to produce neuronal regenerative properties of the cells. As used herein, the term " "Enhancing the neural regeneration properties of the cells" refers to the ability to enhance the proliferation of neural progenitor cells in fibroblasts. and culturing existing neural progenitor cells with one or more agents that protect the cells from apoptosis. For example, fibroblasts have been shown to have enhanced nerve regeneration properties (see below). Fibroblasts can be cultured in aFGF, which allows for the growth of fibroblasts. The method shown involves the selection of fibroblasts expressing the markers CD31 and / or CD73 for the purpose of "neural regeneration" The method further includes the step of optionally priming the subject with one or more agents that can "promote

[0015] In some embodiments of the disclosed methods or compositions, the culture of the fibroblasts comprises culturing the fibroblasts. In some embodiments, the fibroblasts are in a medium that allows for proliferation of the fibroblasts. When these or a combination of these are cultured, activin A, adrenomedrin, aFGF, ALK 1. ALK5, ANF, angiogenin, angiopoietin-2, angiopoietin -3, angiopoietin-4, bFGF, B61, bFGF-inducible activity, cadherin, CA M-RF, cGMP analog, ChDI, CLAF, claudin, collagen, cGMP Analogue, Collagen, Collagen Receptor, α1-β2, α2-β1, Connexin, Cox -2, ECDGF (endothelial cell-derived growth factor), electrocardiogram, EDM, EGF, EMAP, En Endothelin, endothelial cell proliferation inhibitor, endothelial cell survival factor, endothelial cell division Ecdysphagolipid G-protein-coupled receptor-1 (EDG1), ECG, ECI, EDM, EGF, EMAP, endoglin, endothelin, endostatin, endothelial cell proliferation inhibitor , endothelial cell viability maintenance factor, endothelial differentiation glycosylglycolipid G protein-coupled receptor-1 (ED G1), ephrin, Epo, HGF, TGF-β, PD-ECGF, PDGF, IGF, IL8, growth hormone, fibrin fragment E, FGF-5, fibronectin and fibrinogen Ronectin receptor α, 5, β, 1, factor X, HB-EGF, HBNF, HGF, HUAF, cardiac-derived vascular cell growth inhibitor, Ill, IGF-2, IFN-γ, integrin Phosphoreceptor, K-FGF, LIF, leiomyoma-derived growth factor, MCP-1, macrophage derived growth factor, monocyte-derived growth factor. MD-ECI, MECIF, MM2, MMP3, MM P9, urokinase plasminogen activator, neuropilin (NRP1, NR P2), neurotelin, nitric oxide donor, nitric oxide synthases (NOSs), notch , occludin, zona occludin, oncostatin M, PDGF, PDGF-B, P DGF receptor, PDGFR-β, PD-ECGF, PAI-2, PD-ECGF, PF4 , P1GF, PKR1, PKR2, PPAR-γ, PPAR-γ ligand, phosphodiesterase telase, prolactin, prostacyclin, protein S, smooth muscle cell-derived growth factor Sphingosine-1-phosphate-1 (SIP1), Syk , SLP76, tatikinin, TGF-β, Tie1, Tie2, TGF-β, TGF-β Receptor, TIMP, TNF-α, transferrin, thrombospondin, urokinase , VEGF-A, VEGF-B, VEGF-C, VEGF-D, VEGF-E, VEGF , VEGF 164 , VEGI, EG-VEGF, and combinations thereof. In some embodiments, the fibroblasts are grown in growth medium, conditioned medium, or reprogramming medium. It is cultivated underground.

[0016] In some embodiments, the neuronal regeneration properties include inhibition of neuroinflammation, neuronal progenitor cells, enhancing regeneration of brain cells, inhibiting excitotoxicity, preventing or inhibiting apoptosis of brain cells, or performing any of these functions. In some embodiments, the neuronal regeneration is hippocampal regeneration. In some embodiments, the neuronal progenitor cells are cells present in the dentate gyrus. In some embodiments, the neuronal progenitor cells are cells present in the subventricular zone. In some cases, neuroinflammation is mediated by the inflammatory cytokine TNF-α or the cytokine IL-1, Mediated by the cytokine IL-6.

[0017] In some embodiments, preventing or inhibiting apoptosis involves the administration of IGF-1 or is due to the production of VEGF.

[0018] In some embodiments, the method comprises exposing to one or more Toll-like receptor agonists. The method further comprises activating the fibroblasts by Toll-like receptors. 1 or TLR-2 or TLR-3 or TLR-4 or TLR-5 or is TLR-6 or TLR-7 or TLR-8 or TLR-9, or It can be a combination.

[0019] In some embodiments, the agonist of TLR-1 is Pam3CSK4.

[0020] In some embodiments, the agonist of the TLR-2 agonist is HKLM. .

[0021] In some embodiments, the TLR-3 agonist is Poly:IC.

[0022] In some embodiments, the TLR-4 agonist is LPS, buprenorphine , carbamazepine, fentanyl, levorphanol, methadone, cocaine, morphine, Oxacarbazepine, oxycodone, pethidine, glucuronic acid derived from Cryptococcus Lonoxylomannan, morphine-3-glucuronide, lipoteichoic acid, β-defensin 2 These are small molecular weight hyaluronic acid, fibronectin EDA, snapin, and tenascin C. do.

[0023] In some embodiments, the TLR-5 agonist is flagellin.

[0024] In some embodiments, the agonist of TLR-6 is FSL-1.

[0025] In some embodiments, the TLR-7 agonist is imiquimod.

[0026] In some embodiments, the TLR8 agonist is ssRNA40 / LyoVec. do.

[0027] In some embodiments, the agonist of TLR-9 is a CpG oligonucleotide, OD N2006, or agatolimod.

[0028] In some embodiments, the addiction is to heroin, Oxycontin, or Vicodin. Which opioid addiction is the most common? The opioids may or may not be prescription opioids. Opioids may be natural opiates that are alkaloids, including the opium poppy. Natural opiates include morphine, These include codeine, thebaine, and opioids. Opioids are made in the laboratory from natural opiates. The opioids may be semi-synthetic / artificial opioids. Semi-synthetic opioids include hydromorphone, Hydrocodone, oxycodone (prescription drug OxyContin), and drugs made from morphine Heroin. Opioids include fentanyl, pethidine, levorphanol, and methadone. Completely synthetic / artificial opioids, including methadone, tramadol, and dextropoxyphene. In some embodiments, the addiction is to heroin.

[0029] In some embodiments of the disclosed methods or compositions, the fibroblasts are It may be autologous or allogeneic or xenogeneic to the individual being treated.

[0030] In some embodiments of the disclosed methods or compositions, the subject is also administered memantine. Memantine can be administered simultaneously with, before or after the administration of the fibroblasts. Various doses include, but are not limited to, 5 to 100 mg / day or 10 to 30 mg / day. In some embodiments, the subject may also be administered a painkiller, naloxone or analgesic. Compatase or other is administered.

[0031] In some embodiments of the methods of the present disclosure, the fibroblast or cells are encapsulated. In some embodiments, the membrane, polymer capsule, alginate-poly-L-lysine Poly-L-lysine alginate microcapsules, poly-L-lysine barium alginate capsules, alginate Barium phosphate capsule in polyacrylonitrile / polyvinyl chloride (PAN / PVC) It is encapsulated by hollow fibers or polyethersulfone (PES) hollow fibers.

[0032] Further embodiments of the present disclosure include methods for treating brain damage associated with opioid addiction, including the administration of therapeutic cells. A method of treating a wound, wherein the therapeutic cells comprise the steps of: a) obtaining fibroblasts; Cultivating the fibroblasts in a liquid medium capable of allowing the proliferation of the fibroblasts. c) extracting from the single cell suspension cells expressing the markers CD31 and / or CD73. and d) treating said cells with an agent capable of enhancing the production of neuronal regenerative properties of said cells. The cells are optionally priming.

[0033] Further embodiments of the present disclosure include placing the fibroblasts in the subgranular zone (SGZ) of a subject. By this, fibroblasts expressing the CD31 and / or CD73 markers or activated and a method for treating associated hippocampal damage in a subject, comprising administering to the subject an endothelial cell line comprising: Regarding the law.

[0034] Further embodiments of the present disclosure include a method for producing a fibroblast comprising the steps of obtaining fibroblasts and expanding said fibroblasts. and culturing said fibroblasts in a medium that allows for the extraction of single cell suspension cells. and isolating fibroblasts expressing the markers CD31 and / or CD73. administering to a subject a composition comprising a fibroblast and a CD3 and expressing the CD73 marker. This relates to a method for

[0035] Further embodiments of the present disclosure include administering fibroblasts to a subject to induce fibrosis in the subject. In a method for treating brain damage associated with opioid addiction, the cells are In some embodiments, the cells express 73 markers. and b) culturing said fibroblasts in a medium that allows proliferation of said fibroblasts. c) extracting from single cell suspension cells; and d) detecting the markers CD31 and / or and isolating fibroblasts that express CD73. In some embodiments, the cells are obtained by the steps of: a) obtaining fibroblasts; b) expanding said fibroblasts; c) culturing said fibroblasts in a liquid medium capable of detecting the marker C; and d) extracting from said single cell suspension cells that express D31 and / or CD73; The cells can be optionally plated with agents capable of enhancing the production of neuronal regenerative properties of the cells. The neuronal regenerative properties are produced by a process of imitating the neuroinflammation of b) enhancing the renewal of neuronal progenitor cells; c) inhibiting excitotoxicity; and d) may be to prevent apoptosis of brain cells.

[0036] Further embodiments of the present disclosure include fibroblasts or The present invention relates to a pharmaceutical composition for treating addiction, comprising a plurality of such cells. and b) culturing said fibroblasts in a liquid medium capable of allowing proliferation of said fibroblasts. c) culturing fibroblasts expressing markers CD31 and / or CD73; and d) extracting from said single cell suspension cells the production of neuronal regeneration properties of said cells. and optionally priming said cells with an agent capable of enhancing It is possible.

[0037] Any method or composition described herein may be used in combination with any other method or composition described herein. It is contemplated that the present invention may be implemented in terms of articles and that different embodiments may be combined. will be done.

[0038] The use of one or more of the compositions may be used in accordance with any of the methods described herein. Other embodiments are discussed throughout the disclosure. Any embodiment described herein also applies to other aspects of the disclosure, and vice versa. The embodiments in the Examples section are applicable to all aspects of the technology described herein. It is understood that this is a possible embodiment.

[0039] The foregoing describes the features and technical advantages of the present disclosure in order that the detailed description that follows may be better understood. Additional features and advantages which form the subject of the claims of this specification. The disclosed concepts and specific embodiments accomplish the same objectives of the present design. Those skilled in the art will appreciate that the present invention may be readily utilized as a basis for modifying or designing other structures for Such equivalent constructions are to be understood as falling within the scope of the appended claims. It should be understood by those skilled in the art that the present invention is not to depart from the spirit and scope of the invention as described herein. Further objects and advantages of the present invention are set forth in the following detailed description of the present invention: and, both as to construction and method of operation, when considered in conjunction with the accompanying drawings, It will be better understood from the following description. However, the figures are for purposes of illustration and explanation only. It should be expressly understood that these are provided solely for the purposes of illustration and are not intended as a definition of the limits of the present disclosure. be.

[0040] Other objects, features and advantages of the present invention will become apparent from the following detailed description. However, the detailed description of the invention, specific examples and preferred embodiments are given by way of example. However, various changes and modifications that become apparent to those skilled in the art from this detailed description are intended to be included within the scope of the present invention. It should be understood that the present invention is within the spirit and scope of the present invention.

[0041] Fibroblasts and / or their derivatives for inhibiting and / or treating addiction, such as opioid addiction Methods and compositions including products derived therefrom are disclosed. In one embodiment, the method comprises administering to the patient a therapeutically effective amount of an opioid addiction treatment. The patient is administered a population of fibroblasts in a concentration sufficient to inhibit addiction-related brain damage. In one embodiment, fibroblasts are used to promote neuronal regeneration to overcome addiction-related brain changes. In some embodiments, the present invention is used to stimulate hippocampal regeneration following intoxication-induced injury. Regarding intense.

[0042] In certain embodiments, the method comprises administering fibroblasts and / or products derived therefrom. and methods that can be used to inhibit and / or treat addiction, such as opioid addiction. Current embodiments relate, inter alia, to compositions and methods relating to the treatment of addiction and substance abuse. In some embodiments, the compositions and methods include fibroblasts and / or products derived therefrom. In some embodiments, the present invention relates to the treatment or inhibition of opioid addiction by administration of The drug was administered to patients with methadone addiction in concentrations sufficient to inhibit addiction and / or associated brain damage. In some embodiments, the fibroblasts are administered to inhibit addiction-related brain changes. In some embodiments, the method is used to provide neuronal regeneration to overcome the The disclosure also relates to methods for stimulating hippocampal regeneration following addiction-induced damage. The present invention provides the use of fibroblasts as a cell therapy for the treatment of cancer. The cells may be autologous, allogeneic or xenogeneic.

[0043] The intended meanings of terms are defined herein below. I. Definition

[0044] "Opioid addiction" or "opioid use disorder" refers to the harmful consequences of repeated opioid use. The results include patterns of compulsive opioid use and, occasionally, symptoms of tolerance and / or withdrawal. It refers to a condition characterized by physiological dependence on the drug.

[0045] "Drug withdrawal" refers to the sudden cessation or abrupt onset of taking a drug or recreational drug. As a result, "opioid withdrawal" refers to a group of symptoms that occur when the If you suddenly stop or reduce your intake of opioids or This refers to a group of symptoms that occur when taking a drug. Withdrawal symptoms may also occur at intervals between doses. Opioid withdrawal symptoms include anxiety, depression, sweating, vomiting, and diarrhea, muscle cramps, and agitation. Symptoms include insomnia, yawning, dilated pupils, goose bumps, abdominal cramps, runny nose, and increased tear production. Examples include, but are not limited to:

[0046] The terms "substantially the same" or "not significantly different" mean something that is significantly different from what it is being compared to. Alternatively or in combination, the term substantially the same refers to a level of expression that is not refers to an expression level that differs by less than 2, 1.5, or 1.25 fold from the expression or activity level observed in vinegar.

[0047] The terms "subject," "individual," and "patient" are used interchangeably herein. In particular, animals include vertebrates (e.g., primates, mammals, or humans). Mammals include horses, horses, These include cattle, sheep, mice, rats, monkeys, humans, farm animals, sport animals and pets. The following are intended to be included in the scope of the invention: subjects involved in clinical research studies who do not show clinical signs of disease; subjects involved in epidemiological studies; It is an object used as a target or control.

[0048] "Diagnosis" means the process of attempting to determine or identify a possible disease or disorder, or From a statistical point of view, a diagnostic procedure can refer to the opinion reached by this process. It may include a test of the same kind.

[0049] "About" and "approximately" generally refer to the measured value given the nature or precision of the measurements. It means an acceptable degree of error for a quantity. Typically, an exemplary degree of error is given. Within 20 percent (%), preferably within 10%, and more preferably within 5% of a value or range of values. Alternatively, particularly in biological systems, the terms "about" and "approximately" may be used interchangeably. , within an order of magnitude, preferably within 5-fold, more preferably within 2-fold of the given value. In some embodiments, the numerical values ​​discussed herein may be used in conjunction with the term It is contemplated that it can be used in conjunction with "approximately" or "approximately."

[0050] As used herein, the term "comprising" means the inclusion of the recited elements but not the exclusion of others. The term "compositions and methods" is intended to mean that the compositions and methods are not When used in a statement, it is not to the exclusion of other elements of any essential importance for the stated purpose. "Consisting essentially of" in the context of the pharmaceutical compositions of the present disclosure means is intended to include all of the active agents and to exclude any additional unlisted active agents. However, it does not exclude other ingredients of the composition that are not active ingredients. A composition consisting essentially of the defined elements is free of trace contaminants from isolation and purification processes, and Pharmaceutically acceptable carriers such as phosphate buffered saline, preservatives, etc. are not excluded. " is a comprehensive list of trace elements, other ingredients and substantial method steps for administering the compositions of the present invention. A process step or steps to produce a composition or achieve an intended result. The embodiments defined by each of these transition terms are It is within the scope of the present invention to include embodiments described in the context of the term "comprising" " or "consisting essentially of."

[0051] The terms "protein," "polypeptide," and "peptide" refer to gene products or functional proteins. The terms are used interchangeably herein when referring to a protein.

[0052] The terms "ameliorate," "inhibit," or "reduce," or any combination of these terms Reation, when used in the claims and / or specification, refers to a desired result. This includes any measurable reduction or complete inhibition to achieve the above.

[0053] The term "inhibitor" refers to a protein, process (e.g., a metabolic process), or biochemical pathway. It refers to a therapeutic agent that indirectly or directly inhibits the activity or expression of

[0054] The term "agonist" refers to an agent that acts by interacting with a receptor, such as an opioid receptor, to activate the receptor. receptor, thereby initiating the physiological or pharmacological response characteristic of that receptor. Describe the part or drug that can be used.

[0055] As used herein, a "partial agonist" refers to an agonist that binds to a given receptor and agonists activate the receptor but have only partial efficacy at the receptor compared to full agonists. The compound is a moiety or drug having the function of

[0056] As used herein, an "antagonist" is an agent that binds to the receptor at the same site as an agonist. It competitively binds to the receptor but activates the intracellular response initiated by the activated form of the receptor. and thereby capable of inhibiting the intracellular response by an agonist or partial agonist. Please write:

[0057] The term "pharmaceutical formulations" includes salts, solvates, and hydrates of the compounds described herein. Compositions or mixtures of compositions containing at least one active ingredient, including but not limited to is intended to mean

[0058] As used herein, "treat," "treatment," or "treatment" refers to a procedure that provides benefit or benefit. This is an approach to achieve a desired clinical outcome. This includes reduction or relief of symptoms, reduction of pain, or reduction in the frequency of withdrawal symptoms and / or reduction in the occurrence of anxiety or depression and / or Additionally, these terms include a reduction in the severity of a condition or disease. The term "treatment" is intended to encompass the cure and amelioration of one or more of the symptoms of opioid use disorder, for example. In these cases, response to treatment includes cessation of opioid use or at least one This includes cessation of opioid withdrawal symptoms.

[0059] The term "therapeutically effective amount" refers to an amount of cells that treats or inhibits addiction or withdrawal symptoms in a subject. In some embodiments, a therapeutically effective amount is at least or at most or exactly the same as the onset of a symptom. 100, 99, 98, 96, 94, 92, 90, 85, 80, 75, 70, 65, 60 , 55, 50, 40, 30, 20, or 10%, or any derivable range therein. Inhibits.

[0060] The terms "administered" or "administering" as used herein refer to a composition. Any method of providing a composition to an individual such that the composition has its intended effect on the patient. For example, one method of administration is via a medical device such as a catheter, applicator gun, or syringe. The second step of administration may be by an indirect mechanism, such as, but not limited to, using a device. An exemplary method is by a direct mechanism, such as local tissue administration.

[0061] As used herein, "allogeneic" refers to a gene that is derived from one or more individuals of the same species, but Alternatively, the antigen may be immunologically incompatible in its natural setting. It refers to tissues or cells derived from the body.

[0062] As used herein, the term "allograft" refers to the transfer of an organ from a donor to a recipient. The term refers to transplantation of organs, tissues, and / or cells, in which the donor and recipient are different individuals. Cells or tissues transplanted by such procedures are allogeneic This is called a graft or allograft.

[0063] As used herein, "autologous" refers to tissue derived from or derived from the same individual body. Transplantation refers to tissues or cells transferred from a donor (i.e., autologous blood donation; autologous bone marrow transplant).

[0064] As used herein, the term "autologous transplant" refers to the transplantation of a part of the body from an individual. to another part, to the same individual (i.e., the donor and recipient are the same individual); This refers to the transplantation of organs, tissues, and / or cells in a patient with a chronic condition, such as a heart attack, stroke, or stroke. The tissue that is transplanted is called an autologous graft or autologous transplant.

[0065] The term "biologically active" refers to any compound that has a structural, regulatory or biochemical function. For example, biological activity refers to the ability of a molecule to function as a protein in a cell that lacks, for example, a protein activity. Cells lacking a protein activity can be identified by a number of methods, including the restoration of wild-type growth. Complementation can be achieved by, for example, point mutations and frameshift mutations. Cells lacking protein activity are transformed with an expression vector expressing the protein, a derivative thereof, or a portion thereof. In other cases, this is achieved by transfecting a gene product. Fragments (e.g., proteins) have reduced, but not complete, activity of the full-length gene product. may be at detectable levels but may retain the activity of the full-length gene product. A compound is considered to be biologically active (or sometimes referred to as functionally active) if it is be).

[0066] Use of the term "or" in the claims is expressly intended to refer to alternatives only. Unless otherwise indicated, "and / or" is used to mean or alternatives are mutually exclusive. Although the disclosure supports a definition that refers only to alternatives and "and / or," Any limitations discussed with respect to one embodiment may apply to any other embodiment of the invention. It is specifically contemplated that any of the compositions of the present invention may be used in any of the methods of the present invention. and any method of the invention may be used to produce or utilize any composition of the invention. Additionally, the embodiments discussed in the Examples may be used in conjunction with any of the methods discussed herein. The present invention may be applied in the context of any other embodiment.

[0067] When used in the claims and / or this specification in conjunction with the term "comprises," The use of the terms "a" or "an" can mean "one," but it can also mean "one or more." , "at least one" and "one or more." II. Various Embodiments

[0068] Embodiments of the present disclosure are directed to the prevention and treatment of drugs, narcotics, alcohol, marijuana, hallucinogens (mushrooms, PCP , LSD, etc.), inhalants (such as paint thinner or glue), sedatives, sleeping pills, Anti-anxiety drugs, tranquilizers, cocaine, methamphetamines, stimulants of any kind, tobacco Addiction or substance abuse of any kind, including sex, gambling, or a combination thereof It relates to the treatment or prevention of.

[0069] In one embodiment, fibroblasts, including those with regenerative properties, are used to treat any type of nervous system disorder. Used to help people overcome addiction or substance abuse (sometimes with chemical and (and to provide nanoparticle supports). It is known that this is a tendency towards recidivism. Dopamine (DA) transmission in the nucleus accumbens (NA) DA receptor agonists are involved in reward processes. They are self-administered and modulate opioid-seeking behavior. However, D1DA antagonists in NA reduce the reinforcing potency of opioids. Glutamate transmission in A is associated with behavioral sensitization, whereas AMPA receptor inhibition is associated with sensitization. Prevents both the development of sensitization and the increase in glutamatergic transmission following opioid administration in stimulated rats Behavioral sensitization to psychomotor stimulants correlates with abnormalities in the mesoaccumbens dopamine (DA) system. These include at least one DA autoreceptor subsensitivity in the ventral tegmental area. and hypersensitivity of D1 receptors in the nucleus accumbens.

[0070] Animal experiments have demonstrated that NMDA antagonists (noncompetitive and competitive) and AMPA antagonists Both treatments prevented both opioid sensitization and receptor changes in the medial prefrontal cortex and the medial medial prefrontal cortex. Glutamate transmission to the nucleus accumbens DA system was important for the induction of opioid sensitization and receptor correlation Glycine-binding site NMDA antagonists and nitric oxide synthase (NOS) inhibitors reported that it attenuates the development of morphine tolerance and even reverses established tolerance or dependence. It has been reported that glutamate antagonists can inhibit tolerance to morphine without exerting its analgesic effect. Regulation of sex and addiction is associated with adaptive changes mediated by the NMDA / NO cascade Suggesting prevention of neuronal plasticity. Expresses both NMDA and mu opioid receptors. In neurons that The magnitude of the current is enhanced. Mu receptor activation is suppressed by removing the Mg+ blockade. It functions as a cascade that increases NMDA activation and allows for the subsequent production of NO. Cade alters gene expression, induces neuroplasticity, and leads to both tolerance and dependence. The latter neurochemical event reduces the analgesic cascade effect of morphine. N-methyl-D-aspartate (NMDA) antagonists inhibit mu-opioid stimulation It is possible to prevent the phenomenon of drug tolerance without directly affecting the analgesic effect of

[0071] In one embodiment, the present disclosure provides a method for reducing dependency, in part, by modulating NMDA transmission. This includes the use of fibroblasts as a therapeutic cell source to reduce opioid addiction and chronic Drug tolerance, dependence, addiction, and withdrawal symptoms in both sexual and chronic pain patients are thought to be due to the NMDA receptor complex. It may be partially mediated by the body. In animal studies, glycine site receptor antagonists has been shown to be effective in reducing withdrawal symptoms and eliminating opiate drug addiction In contrast, memantine results in eliminating symptoms of drug withdrawal and addiction in animal studies. Results were inconsistent and usually negative, which may reflect inadequate duration of treatment. Glycine site antagonists administered to chronic heroin and cocaine users An unexpected finding was the gradual reduction in addiction, tolerance, and dependence, resulting in Discontinuation of treatment did not result in any evidence of relapse, and patients remained drug-free for several years.

[0072] In one embodiment of the present disclosure, fibroblasts are used to obtain a synergistic effect of reducing toxicity. It is administered together with memantine at a dose of 5 to 100 mg / day, advantageously 10 to 30 mg / day. Chronically administered oral doses (serum levels ranging from 0.25 to 2.0 μg / ml) Mantine is effective in treating acute and chronic opioid tolerance. The combination of memantine and analgesics reduces the likelihood of opioid tolerance and physical dependence. Fibroblasts may be used in combination with current standard drug therapy (i.e., naloxone or acolinyl chloride). Memantine can be administered in combination with cyclosporine (cyclosporine amptase) to patients with chronic tolerance and dependence. Both have been proposed in the treatment of illicit drug addiction. Intravenous administration of memantine in the event of an overdose has also been proposed. The fibroblast composition of the present invention is administered with memantine, naloxone, acopatase or other compounds. Together with the disclosed fibroblast compositions, about, at least about, or at most about 0.01, 0. 02, 0.03, 0.04, 0.05, 0.06, 0.07, 0.08, 0.09, 0. 1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1.0, 1. 1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2.0, 2. 1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.8, 2.9, 3.0, 3. 1, 3.2, 3.3, 3.4, 3.5, 3.6, 3.7.3.8, 3.9, 4.0, 4. 1, 4.2, 4.3, 4.4, 4.5, 4.6, 4.7, 4.8, 4.9, 5.0, 5. 1, 5.2, 5.3, 5.4, 5.5, 5.6, 5.7, 5.8, 5.9, 6.0, 6. 1, 6.2, 6.3, 6.4, 6.5, 6.6, 6.7, 6.8, 6.9, 7.0, 7. 1, 7.2, 7.3, 7.4, 7.5, 7.6, 7.7, 7.8, 7.9, 8.0, 8. 1, 8.2, 8.3, 8.4, 8.5, 8.6, 8.7, 8.8, 8.9, 9.0, 9. 1, 9.2, 9.3, 9.4, 9.5, 9.6, 9.7, 9.8, 9.9, 10.0, 1 0.5, 11.0, 11.5, 12.0, 12.5, 13.0, 13.5, 14.0, 1 4.5、15.0、15.5、16.0、16.5、17.0、17.5、18.0、1 8.5、19.0.19.5、20.0、1、2、3、4、5、6、7、8、9、10、 11、12、13、14、15、16、17、18、19、20、21、22、23、2 4、25、26、27、28、29、30、31、32、33、34、35、36、37 、38、39、40、41、42、43、44、45、46、47、48、49、50、 51、52、53、54、55、56、57、58、59、60、61、62、63、6 4、65、66、67、68、69、70、71、72、73、74、75、76、77 、78、79、80、81、82、83、84、85、86、87、88、89、90、 91、92、93、94、95、96、97、98、99、100、105、110、1 15、120、125、130、135、140、145、150、155、160、1 65、170、175、180、185、190、195、200、205、210、2 15、220、225、230、235、240、245、250、255、260、2 65、270、275、280、285、290、295、300、305、310、3 15、320、325、330、335、340、345、350、355、360、3 65、370、375、380、385、390、395、400、410、420、4 25、430、440、441、450、460、470、475、480、490、5 00、510、520、525、530、540、550、560、570、575、5 80、590、600、610、620、625、630、640、650、660、6 70, 675, 680, 690, 700, 710, 720, 725, 730, 740, 7 50, 760, 770, 775, 780, 790, 800, 810, 820, 825, 8 30, 840, 850, 860, 870, 875, 880, 890, 900, 910, 9 20, 925, 930, 940, 950, 960, 970, 975, 980, 990, 1 000, 1100, 1200, 1300, 1400, 1500, 1600, 1700, 1 800, 1900, 2000, 2100, 2200, 2300, 2400, 2500, 2 600, 2700, 2800, 2900, 3000, 3100, 3200, 3300, 3 400, 3500, 3600, 3700, 3800, 3900, 4000, 4100, 4 200, 4300, 4400, 4500, 4600, 4700, 4800, 4900, 5 000, 6000, 7000, 8000, 9000, 10000 milligrams (mg) Or micrograms (mcg) or μg / kg or micrograms / kg / min or mg / kg / min or micrograms / kg / hour or mg / kg / hour, or Any range derived therefrom may be administered.

[0073] In some embodiments, the present invention provides a method for overcoming addiction-related hippocampal damage. We demonstrate that fibroblast administration can help the brain generate new neural stem / progenitor cells (NSPCs) during adulthood. The hippocampus is one of the most plastic regions of the brain, and the dentate gyrus Granular cells are born in adulthood. The precursors of these cells form in the interstitial space between the hilus and the granular cell layer. One of the characteristics of adult hippocampal neurons is that It is the specific electrophysiological capacity for the extreme changes required during the early stages of maturation. The properties of SGZ are important for memory formation and further physiological functions. In one embodiment, the fibroblasts provide a suitable niche for the proliferation and differentiation of stem cells. Administration of cells or activated fibroblasts increases the proliferation of SGZ cells to induce neuronal repair. It can stimulate and in some cases induce the formation of memories that are not related to the addictive memory. It is carried out in such a manner.

[0074] In another embodiment, the fibroblasts and / or activated fibroblasts are in the environment of the SGZ cells. For example, fibroblasts can be used to regulate astrocyte-mediated inflammatory responses. It is administered to reduce the production of inflammatory mediators and toxins. Astrocytes, as a cellular component, express Wnt proteins and Sonic hedgehog (Shh). In part through the release of molecular signals such as It is known in the art that oxidative stress plays an active role in the regulation of oxidative stress. In embodiments, the fibroblasts are provided with a method for overcoming opioid-related changes in fibroblast activity. For example, fibroblasts are administered to promote neuronal survival, immune responses, and neurotransmission. It has been shown to play an essential role in the regulation and metabolism of substances. Or each stimulant that can affect those niches in the hippocampus may have a role in memory and behavior. Many studies have shown that depression, schizophrenia, bipolar disorder, addiction, and other disorders can be significantly altered. Alterations in adult hippocampal neurogenesis have been demonstrated in neuropsychological disorders. In one embodiment, the present disclosure provides a method for the treatment of fibroblastic leukemia and / or myocardial infarction (CI)-related inflammatory bowel disease (CI-DMF) in patients with rheumatoid arthritis. The present invention includes administration of the cells.

[0075] In some embodiments of the invention, the therapeutic agent for producing the protein / peptide is Therapeutically effective fibroblasts are activin A, adrenomedrin, aFGF, ALK1, A LK5, ANF, angiogenin, angiopoietin-2, angiopoietin-3, Angiopoietin-4, bFGF, B61, bFGF-inducible activity, cadherin, CAM-R F, cGMP analog, ChDI, CLAF, claudin, collagen, cGMP analog , collagen, collagen receptor, α1-β2, α2-β1, connexin, Cox-2, ECDGF (endothelial cell-derived growth factor), electrocardiogram, EDM, EGF, EMAP, endothelial Endothelin, endothelial cell proliferation inhibitor, endothelial cell survival factor, endothelial cell differentiation factor Engolipid G-protein-coupled receptor-1 (EDG1), ECG, ECI, EDM, EGF , EMAP, endoglin, endothelin, endostatin, endothelial cell proliferation inhibitor, endo Endothelial cell viability factor, endothelial differentiation glycosylglycolipid G protein-coupled receptor-1 (EDG1) , ephrin, Epo, HGF, TGF-β, PD-ECGF, PDGF, IGF, IL8 , growth hormone, fibrin fragment E, FGF-5, fibronectin and fibronectin chin receptor α 5 β 1, factor X, HB-EGF, HBNF, HGF, HUA F, cardiac vascular cell growth inhibitor, Ill, IGF-2, IFN-γ, integrin receptor receptor, K-FGF, LIF, leiomyoma-derived growth factor, MCP-1, macrophage-derived growth factor Growth factor, monocyte-derived growth factor. MD-ECI, MECIF, MM2, MMP3, MMP9, Urokinase plasminogen activator, neuropilins (NRP1, NRP2) , Neurotelin, Nitric oxide donor, Nitric oxide synthases (NOSs), Notch, Oct Ludin, Zona occludin, Oncostatin M, PDGF, PDGF-B, PDGF Receptor, PDGFR-β, PD-ECGF, PAI-2, PD-ECGF, PF4, P1 GF, PKR1, PKR2, PPAR-γ, PPAR-γ ligand, phosphodiesterase Ze, prolactin, prostacyclin, protein S, smooth muscle cell-derived growth factor, Smooth muscle cell-derived chemotactic factor. Sphingosine-1-phosphate-1 (SIP1), Syk, SL P76, tatikinin, TGF-β, Tie1, Tie2, TGF-β, TGF-β receptor , TIMP, TNF-α, transferrin, thrombospondin, urokinase, VE GF-A, VEGF-B, VEGF-C, VEGF-D, VEGF-E, VEGF, VE GF 164 VEGI, EG-VEGF, and combinations thereof. The cells are cultured in a manner that produces proteins / peptides that cannot be detected by conventional methods.

[0076] In the context of this disclosure, references to "CD45," "CD34," and "CD31" are References to all forms of these molecules, as well as functional fragments, mutants or variants thereof. In addition, alternative splicing of CD45, CD34, and CD31 mRNAs or any isoforms that may arise from isomers or polymorphisms of these molecules. This is a reference to.

[0077] Reference to a "phenotypic profile" refers to the transcription and / or expression of genes encoding subject markers. or the expression product translated therefrom. Most cells falling within the scope of the claimed fibroblast population are surface-bound. Characterized by the presence or absence of a test marker as a constant expression product, within a defined population Some cells entering the cell line have upregulated transcription of a given marker, but may not result in a cell surface-anchored expression product. It should be recognized that a new classification may initially show changes only at the system level. The progression of cells to the differentiation stage is not yet clear in relation to changes in expression product levels. show transient gene expression changes. However, these cells nevertheless Within the claimed cell populations, they are not necessarily associated with cell surface marker expression. In some embodiments of the present invention, the Fibroblasts expressing CD31 and CD73 in morphology can be used for the treatment of addiction. will be done.

[0078] Some embodiments are based on expression of CD73 or lack of expression of certain proteins. and a method for selecting fibroblasts for enhanced efficacy in the treatment of opioid addiction. Various terms are used to describe cells in culture. " is usually taken from a living organism and grown under controlled conditions ("culture" or "incubation"). "Primary cell culture" refers to cells, tissues, or organs directly harvested prior to the first subculture. It is a culture of cells that are placed in a culture medium and cultured under conditions that promote cell growth and / or division. This increases the size of the cell population, and is used to grow cells in culture. is sometimes measured by the time it takes for the number of cells to double. A "cell line" is a cell that has been generated by one or more subcultures of a primary cell culture. Each round of subculture is called a passage. When they are cultured, they are called passaged. A particular cell population, or cell line, is sometimes called The term "culture" refers to or is characterized by the number of times that a culture has been passaged. For example, a culture that has been passaged 10 times The cell population can be referred to as a P10 culture. Primary culture, i.e., isolating cells from tissue The first culture after the first subculture is designated as P0. Following the first subculture, the cells are subcultured (P1 or After the second subculture, the cells become tertiary cultures (P2 or passage 2). During the period of passaging, there may be many population doublings; therefore, It will be understood by those skilled in the art that the number of population doublings of an organism is greater than the number of passages. The proliferation of cells in vitro (i.e., the number of population doublings) is a function of the seeding density, substrate, medium, growth conditions, and It depends on many factors, including but not limited to, the time between passaging.

[0079] "Conditioned medium" is the medium in which a particular cell or cell population has been cultured and then removed. When cells are grown in culture, they secrete cellular factors that can provide trophic support to other cells. Trophic factors are substances that promote or at least support the survival, growth, proliferation and / or maturation of cells. These trophic factors are substances that stimulate or increase the activity of cells. , cytokines, extracellular matrix (ECM), proteins, vesicles, antibodies, and granules. The medium containing the cellular factors is a conditioned medium. In some embodiments of the compositions or methods of the invention, trophic factors are used. In embodiments, the compositions and methods include fibroblast-derived products (e.g., fibroblasts) of the present disclosure. It is also possible to collect supernatant from regenerative cells such as fibroblasts. Preferably, the supernatant and / or components thereof are useful for providing factors for the treatment of opioid addiction. In some embodiments, fibroblasts are used as a source of fibroblasts from opioid addiction. Factors that aid in recovery (e.g., interleukin-1 receptor antagonists, interleukin Interleukin-10, Interleukin-20, Interleukin-35, Interleukin- 37, TGF-β, BDNF, and / or NGF) to enhance production. , are transfected with one or more genes.

[0080] Any of the compositions or methods of the present disclosure are intended to include cell encapsulation. In some embodiments, the cells are individually encapsulated. In some cases, cells are removed after transplantation. In embodiments, a relatively large configuration that encapsulates many cells, such as within a single membrane, is preferred for collection. It may be used to provide a convenient means for the microencapsulation of stem cells. In various embodiments, a wide variety of materials may be used. For example, polymer capsules, alginate-poly-L-lysine-alginate microcapsules, Capsules, poly-L-lysine barium alginate capsules, barium alginate capsules , polyacrylonitrile / polyvinyl chloride (PAN / PVC) hollow fibers, and polyethylene Examples of the hollow fibers that can be used for the administration of stem cells include polyethersulfone (PES) hollow fibers. Techniques for microencapsulation are known to those skilled in the art and are described, for example, in Chang, P. .et al., 1999;Matthew, H.W. et al., 1991;Yanagi, K. et al., 19 89; Cai ZH et al., 1988; Chang, TM, 1992 and US Patents No. 5, 639, 275 (which is used, for example, to stabilize biologically active molecules). We describe a biocompatible capsule for the long-term maintenance of cells expressing GFP. The method is described in European Patent Publication No. 301,777 and U.S. Pat. No. 4,933,363. , 353, 888;4, 744, 933;4, 744, 933;4, 749, 620;4 , 814, 274;5, 084, 350;5, 089, 272;5, 578, 442;5 , 639, 275; and 5, 676, 943. All of the above relate to stem cell encapsulation. In applicable part, the disclosures herein are incorporated by reference.

[0081] Certain embodiments of the present disclosure involve the incorporation of stem cells into a polymer (e.g., a biopolymer or a synthetic polymer). Examples of biopolymers include fibronectin, fibrin, and fibrino These include, but are not limited to, erythrocyte sedimentation proteins, thrombin, collagen, and proteoglycans. Other factors (e.g., the cytokines mentioned above) may also be incorporated into the polymer. In another embodiment of the present disclosure, stem cells may be incorporated into the interstices of the three-dimensional gel. Large polymers or gels are typically surgically implanted. The polymers or gels that can be formulated are suitable for use in other common, more convenient, non-surgical routes. It can be administered by

[0082] When referring to cultured vertebrate cells, the term senescence (also replicative senescence or cellular senescence) is used a property attributed to cell culture; i.e., the ability to proliferate beyond a finite number of population doublings Senescence is the process by which cells become cellular and are transformed into fibroblast-like cells. Although first described using endothelial cells, most normal human cell types that can be grown successfully in culture are Different cell types undergo cellular senescence. The in vitro life span of different cell types varies, but the maximum life span is typically Fewer than 100 population doublings (this is because all cells in the culture are senescent and therefore (The number of doublings required to make the nutrient indivisible.) Ageing does not depend on chronological time, but rather Growth factors are measured by the number of cell divisions or population doublings that a culture has undergone. Cells rendered quiescent by removal of growth factors are able to proliferate and differentiate when growth factors are reintroduced. Cleavage is allowed to resume and thereafter undergo the same number of doublings as equivalent cells grown continuously. Similarly, cells were frozen in liquid nitrogen after various numbers of population doublings, then thawed, and When cultured, they produce substantially the same number of cells as cells maintained unfrozen in culture. Senescent cells are not dead or dying cells, but are actually undergoing programmed cell division. These cells are resistant to apoptosis and remain non-dividing for up to three years. Senescent cells are highly viable and metabolically active, but do not divide. The non-dividing state of senescent cells is It has not yet been found to be reversible by any biological, chemical, or viral agent. It hasn't been done yet.

[0083] As used herein, the term "growth medium" generally refers to a medium sufficient for the culture of umbilicus-derived cells. In particular, one currently preferred medium for culturing the cells of the invention herein is Particularly preferred are Dulbecco's modified essential medium (also abbreviated herein as DMEM). Of particular interest is DMEM-low glucose (also DMEM-LG) (Invitrogen). DMEM low glucose is preferably 15% (v / v) fetal bovine serum (e.g., defined fetal bovine serum, Hyclone, Logan U) tah), antibiotic / antimycotic (preferably penicillin (100 units / milliliter) , streptomycin (100 milligrams per milliliter), and amphotericin B (0 0.25 micrograms / milliliter), (Invitrogen, Carlsbad, CA), and 0.001% (v / v) 2-mercaptoethanol (Sigma, St In some cases, different growth media are used. or different supplements are provided, and these are usually provided as supplements to the growth medium. As shown in the text.

[0084] In some embodiments of the present disclosure, standard growth conditions are utilized. "Standard growth conditions" refers to 37 days in a standard atmosphere containing 5% CO2. o Cultivate cells at °C. The relative humidity should be maintained at approximately 100%. Although the above conditions are beneficial for culture, Suitable conditions may include those options available to one of skill in the art for culturing cells, such as body temperature, CO2, relative humidity, and the like. Understand that this can be altered by changing humidity, oxygen, growth medium, etc. I want to be.

[0085] In some embodiments of the present disclosure, the conditioned medium used to treat an addiction is decellularized. In some embodiments, the fibroblasts are used to generate the fibroblasts. In some cases, the cells are treated with various dedifferentiation compositions that can result in increased pluripotency. In an embodiment, the fibroblasts are treated with cytoplasm from less differentiated cells. Such cells, commonly known as .gamma.- ... , which is incorporated by reference. Without limitation, the useful versatility of cytoplasmic extractions The cells include parthenogenetic stem cells [9-23], embryonic stem cells [24-25], and induced pluripotent stem cells. [26-30], Stimulus-induced Acquisition of Pluripotency (STAP)

[31] , and Somatic Cell Nuclear Transfer-derived Stem Cells Extraction of cytoplasmic material was performed as described in the art. It is possible to do so.

[0086] In some embodiments of the invention, pluripotent cells are adapted to enter the interphase stage of the cell cycle. The resulting mixture was then extracted using conventional methods. o C: 10 ml of conical tube The cells were washed by centrifugation in a tube at 500 x g for 10 minutes, the supernatant was discarded, and The cell pellet is resuspended in a total volume of 50 ml cold PBS. The cells are centrifuged at 500 x g for 10 min. Between, 4 o Repeat this washing step, centrifuge at 37 °C, and transfer the cell pellet to approximately 20 volumes of ice. Cold phase cell lysis buffer (20mM Hepes, pH8.2, 5mM MgCl2, 1mM M DTT, 10 μM aprotinin, 10 μM leupeptin, 10 μM pepstatin A, 10 μM soybean trypsin inhibitor, 100 μM PMSF, and preferably 20 μ The cells were then centrifuged at 800 x g for 10 min and resuspended in 100 µg / ml cytochalasin B. o Centrifuge at C The cell pellet was then pelleted by separation, the supernatant was discarded, and the cell pellet was mixed with 1 volume or less of interphase cell lysis buffer. Carefully resuspend in buffer. Incubate the cells on ice for 1 hour to allow the cells to swell. The cells were sonicated using a tip sonicator or sonicated using a glass mortar and pestle. Lyse the cells by either homogenization or Dounce homogenization. This was done until at least 90% of the nuclei were lysed, which was assessed using a phase contrast microscope. The sonication time required to lyse at least 90% of the cells and nuclei is: The time required to prepare the extract may vary depending on the type of cells used. centrifuge in a tabletop centrifuge at 10,000-15,000 x g for 15 minutes. , 4 o Centrifuge at 20 C. Remove the tubes from the centrifuge and immediately place on ice. Carefully collect the supernatant using a 10 μl pipette tip and discard from some of the tubes. Pool these supernatants and place on ice. This supernatant is called "interphase cytoplasmic" or "IS15" The cell extract was aliquoted into 20 μL volumes of extract per tube on ice. Immediately flash freeze on liquid nitrogen and store at -80 °C until use. o Can be stored at ℃ Alternatively, centrifuge the cell extracts in ultracentrifuge tubes on ice (e.g., SW55 Ti rotor; Beck If necessary, cover the top of the tube with mineral oil. 0 × g for 3 h, 4 o C. Sediment the membrane vesicles contained in the IS15 extract by centrifugation at C. At the end of the centrifugation, discard the oil. Carefully collect the supernatant, pooling if necessary. Place in a cold 1.5 ml tube on ice. This supernatant is called "IS200" or "interphase cytoplasmic zone". The extract was aliquoted as described for the IS15 extract. If desired, the extract can be enriched with additional nuclear factors. For example, The reprogramming extract may be purified from cells of the cell type from which the reprogramming extract was derived, and purified as described above. The nuclear factors can be dissolved by ultrasonication under stirring at a concentration of 0.15 to 800 mM. Incubate in nuclear buffer containing NaCl or KCl for 10-60 minutes. The lysate is centrifuged to sediment the unextractable components. The supernatant containing the factors is dialyzed to remove NaCl or KCl. The dialyzed nuclear extract is then This nuclear extract is then coated onto a plate and frozen prior to adding nuclei for reprogramming. The whole cell extracts are added at various concentrations.

[0087] Some embodiments of the present disclosure provide alternatives to cell extracts (e.g., reprogramming media). The reprogramming medium may include one or more natural or recombinant factors (e.g., nucleic acids or Proteins (e.g., T cell receptors or other signaling surface molecules, DNA methyltransferases, etc.) transferase, histone deacetylase, histone, nuclear lamin, transcription factor, activator In the embodiment, the inhibitor, repressor, growth factor, hormone, or cytokine is placed in a solution (e.g., a buffer). Preferably, the one or more factors are added to the donor cells. It is specific to the cell type that you want to become.

[0088] In some embodiments, the extract is an inhibitor of reprogramming fibroblasts in culture. In one embodiment, fibroblasts grown on cover slips are cultured with bacteria. Reversibly permeabilize cells with the toxin streptolysin O and expose them to extracts of pluripotent stem cells at 2 mM The cells are resealed with CaCl2 and allowed to grow in culture. In one embodiment, the fibroblasts are RPMI 1640 in 12-well plates, ~100,000 cells / coverslip Cells are grown on 16 mm poly-L-lysine coated coverslips. +2 Includes Hanks Balanced Salt Solution (Gibco-BRL)-free 200 ng / ml streptomycin In ricin O, 37 o Permeabilize in normal atmosphere for 50 min at °C. More than 80% of fibroblasts were permeable under these conditions as judged by uptake. Aspirate the streptolysin O; extract pluripotent stem cells onto the coverslip. Layer 80x1 of material; CO2 in the atmosphere 37 o Incubate at 37 °C for 1 hour. It contained an ATP generating system and 1 mM each of ATP, CTP, GTP, and UTP. Extracts from vesicles are prepared as described above. To reseal the plasma membrane, 2 mM Ca Add RPMI 1640 containing Cl2 (added from a 1M stock in H2O) to the wells. Cells 37 o Incubate at 4 °C for 2 h. This method results in approximately 100% of permeabilized cells. The tube was resealed. RPMI containing Ca +2 was replaced with RPMI and the cells were grown for several weeks. Several descriptions of cytoplasmic transfer have been published and are incorporated by reference [35-37 Once dedifferentiated fibroblasts are obtained, the conditioned medium and / or the fractions derived from said conditioned medium are The exosomes are concentrated and used therapeutically by administration to an intoxicated patient. This can be done intracavitary, intravenous, oral, and / or subcutaneous. Formulation, dosage and route of administration

[0089] An embodiment includes a method for treating opioid addiction comprising administering to a subject a therapeutically effective amount of fibroblasts or a product derived therefrom. This invention includes medicaments, therapeutic compositions, formulations, preparations and related methods.

[0090] The composition may take the form of, for example, a solution, a suspension, or a film, and may be about 10% to about 95% or about 2 In an embodiment, the composition contains 5% to about 70% cells or products derived therefrom. The composition may be administered in any of a number of suitable ways. by intrathecal (intrathecal), locally, by means (e.g., intrathecal, intraventricular), or intravenous It can be administered systemically by means of an aya reservoir. It can be administered transdermally, rectally, vaginally, sublingually, and and other means, including intranasal.

[0091] The composition is administered in a compatible manner that is therapeutically effective, tolerable, and safe. The amount administered depends on the subject being treated. Precise amounts of cells required to be administered will depend on the judgment of the practitioner.

[0092] In many cases, at most about 3, 4, 5, 6, 7, 8, 9, 10, or more (or It may be desirable to have multiple doses of the drug (any range derivable therein). 1 day to multiple weekly intervals of 1, 2, 3, 4, 5, 6, 7, 8, 9, 10 or more weeks (or any range derivable therein). Following a course of administration, for example, Assessment of symptoms, pain, mood, behavior, or catastrophizing may be performed.

[0093] As used herein, "pharmacologically acceptable" or "pharmaceutical acceptable" means There is a composition.

[0094] The dosage of pharmaceutical compositions and formulations will depend on the type of formulation and will vary according to the size and health of the subject. Various combinations and dosages are contemplated and are within the scope of the present invention and are included within the scope of the "pharmaceutical "Compounds that are" or "pharmacologically acceptable" are within the scope of "compounds that are" or "pharmacologically acceptable", e.g., Anywhere between 5 and 100 mg of memantine in combination with the cells or compositions of the present disclosure. The term "pharmaceutical acceptable" means a dosage that is acceptable when administered to an animal or human. In particular, the present invention provides molecular entities and compositions that do not produce adverse, allergic, or other untoward reactions. cormorant.

[0095] Upon formulation, solutions will be administered in a manner compatible with the dosage formulation and to provide therapeutic or prophylactic The therapeutic agent is administered in an amount that is substantially effective. The formulations are easily administered in a variety of dosage forms. The effective amount of a therapeutic or prophylactic composition is determined based on the intended goal. Alternatively, the term "dose" refers to a physically discrete unit suitable for use in a subject, each unit being The compound is capable of producing the desired response as discussed above in relation to its administration, i.e., appropriate route and regimen. The amount to be administered will depend on the number of treatments and the unit dose. The exact amount of the composition will depend on the desired outcome and / or protection, both in terms of the amount and the dosage. Dosage and administration will depend on the judgment of the practitioner and will be peculiar to each individual. Factors affecting dosage include: The physical and clinical condition of the elephant, the route of administration, the intended therapeutic goal (alleviation of symptoms vs. cure), and the efficacy, stability, and toxicity of the particular composition.

[0096] In certain embodiments, the subject is administered memantine, naloxone, or a combination of the fibroblast compositions of the present disclosure. The amount of the acetylcholine, acopatase or other compound is at least about 0.01, 0.02, or 0.03. .02、0.03、0.04、0.05、0.06、0.07、0.08、0.09、0 .1、0.2、0.3、0.4、0.5、0.6、0.7、0.8、0.9、1.0、1 .1、1.2、1.3、1.4、1.5、1.6、1.7、1.8、1.9、2.0、2 .1、2.2、2.3、2.4、2.5、2.6、2.7、2.8、2.9、3.0、3 .1、3.2、3.3、3.4、3.5、3.6、3.7.3.8、3.9、4.0、4 .1、4.2、4.3、4.4、4.5、4.6、4.7、4.8、4.9、5.0、5 .1、5.2、5.3、5.4、5.5、5.6、5.7、5.8、5.9、6.0、6 .1、6.2、6.3、6.4、6.5、6.6、6.7、6.8、6.9、7.0、7 .1、7.2、7.3、7.4、7.5、7.6、7.7、7.8、7.9、8.0、8 .1、8.2、8.3、8.4、8.5、8.6、8.7、8.8、8.9、9.0、9 .1、9.2、9.3、9.4、9.5、9.6、9.7、9.8、9.9、10.0、 10.5、11.0、11.5、12.0、12.5、13.0、13.5、14.0、 14.5、15.0、15.5、16.0、16.5、17.0、17.5、18.0、 18.5、19.0.19.5、20.0、1、2、3、4、5、6、7、8、9、10 、11、12、13、14、15、16、17、18、19、20、21、22、23、 24、25、26、27、28、29、30、31、32、33、34、35、36、3 7、38、39、40、41、42、43、44、45、46、47、48、49、50 、51、52、53、54、55、56、57、58、59、60、61、62、63、 64、65、66、67、68、69、70、71、72、73、74、75、76、7 7、78、79、80、81、82、83、84、85、86、87、88、89、90 、91、92、93、94、95、96、97、98、99、100、105、110、 115、120、125、130、135、140、145、150、155、160、 165、170、175、180、185、190、195、200、205、210、 215、220、225、230、235、240、245、250、255、260、 265、270、275、280、285、290、295、300、305、310、 315、320、325、330、335、340、345、350、355、360、 365、370、375、380、385、390、395、400、410、420、 425、430、440、441、450、460、470、475、480、490、 500、510、520、525、530、540、550、560、570、575、 580、590、600、610、620、625、630、640、650、660、 670、675、680、690、700、710、720、725、730、740、 750、760、770、775、780、790、800、810、820、825、 830、840、850、860、870、875、880、890、900、910、 920、925、930、940、950、960、970、975、980、990、 1000、1100、1200、1300、1400、1500、1600、1700、 1800、1900、2000、2100、2200、2300、2400、2500、 2600、2700、2800、2900、3000、3100、3200、3300、 3400, 3500, 3600, 3700, 3800, 3900, 4000, 4100, 4200, 4300, 4400, 4500, 4600, 4700, 4800, 4900, 5000, 6000, 7000, 8000, 9000, 10000 milligrams (mg) or micrograms (mcg) or μg / kg or micrograms / kg / min or mg / kg / min or micrograms / kg / hour or mg / kg / hour, or is administered within any range derived therefrom.

[0097] Doses are as needed or at 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12 , 18, or 24 hours (or any range derivable therein), or once per day; 2, 3, 4, 5, 6, 7, 8, 9 times, or once per day (or any inducible The drug may be administered at any interval (any range). It may be administered first before or after symptoms of the disease appear. In some embodiments, the patient is on regimen 1, 2, 3, 4, 5, 6, 7, 8, 9, 1 The first dose is administered at 0, 11, 12 hours (or any range derivable therein), or when the patient is in a Experiencing or exhibiting a sign or symptom (or any range inducible therein) 1, 2, 3, 4, or The vaccine will be administered 5 days later. Patients will receive the vaccine on the 1st, 2nd, 3rd, 4th, 5th, 6th, 7th, 8th, and 9th days. , 10 days or more (or any range that can be derivable therein), or symptoms of the condition have disappeared or until relieved or for 6, 12, 18, or 24 days after symptoms have disappeared or been relieved Treatment may be performed hours, or 1, 2, 3, 4, or 5 days later.

[0098] In some embodiments, the subject's treatment includes, for example, 1, 2, 3, 4, 5, 6, or Every 7 days, or every 1, 2, 3, 4, and 5 weeks, or every 1, 2, 3, 4, 5, 6, 7, 8, These treatments may be repeated every 9, 10, 11, or 12 months. The capacity of the device can be increased.

[0099] The patient may receive at least or up to 10 mg of a composition or combination of compounds described herein. , about 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 3 1, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44 , 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 7 1, 72, 73, 74, 75, 76, 92, 93, 94, 95, 96, 97, 98, 99 , or 100 mg / kg (or any range derivable therein).

[0100] The patient may receive at least or up to 10 mg of a composition or combination of compounds described herein. , about 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 3 1, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44 , 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 7 1, 72, 73, 74, 75, 76, 92, 93, 94, 95, 96, 97, 98, 10 0, 110, 150, 160, 170, 180, 190, 200, 210, 220, 23 0, 240, 250, 260, 270, 280, 290, 300, 310, 320, 33 0, 340, 350, 360, 370, 380, 390, 400, 410, 420, 43 0, 440, 441, 450, 460, 470, 480, 490 mg / kg / day (or The amount of the compound that is administered is in the range of any of the ranges derived therefrom. Cell Encapsulation

[0101] In some embodiments, the fibroblasts of the present disclosure may be encapsulated. It is contemplated that any of a number of methods of cell encapsulation may be used. In embodiments, the cells are individually encapsulated. In embodiments where the cells are removed after implantation, the cells are encapsulated in a single membrane. Relatively large structures that encapsulate many cells, such as within a single membrane, provide a convenient means for recovery. In various embodiments for the microencapsulation of stem cells, A variety of materials can be used. Such materials include, for example, polymer capsules. Cells, alginate-poly-L-lysine-alginate microcapsules, poly-L-lysine Barium alginate capsule, Barium alginate capsule, Polyacrylonitrile / Polyvinyl chloride (PAN / PVC) hollow fiber and polyethersulfone (PES) hollow For the microencapsulation of cells, which can be used for the administration of stem cells These techniques are known to those skilled in the art and are described, for example, in Chang, P. et al., 1999; Matt. hew, HW et al., 1991; Yanagi, K. et al., 1989; Cai ZH et al. , 1988; Chang, TM, 1992 and U.S. Pat. No. 5,63 9,275 (This is, for example, for the long-term maintenance of cells stably expressing biologically active molecules. Further methods of encapsulation are described in European Patent Publication No. No. 301,777 and U.S. Pat. No. 4,353,888; 44,933;4,744,933;4,749,620;4,814,274;5,0 84,350; 5,089,272; 5,578,442; 5,639,275; and 5 ,676,943, all of which are incorporated by reference in their entirety with respect to stem cell encapsulation. Certain embodiments involve the incorporation of stem cells into a polymer (e.g., a biopolymer). Examples of biopolymers are fibronectin, fibrinogen, and fibronectin. , fibrinogen, thrombin, collagen, and proteoglycans, Other factors (such as the above-mentioned cytokines) may also be expressed by the polymer. In another embodiment of the present invention, the stem cells can be incorporated into the interstices of the three-dimensional gel. Larger polymers or gels are typically surgically implanted. Small enough Polymers or gels that can be formulated into particles or fibers are another common and more convenient alternative. , can be administered by non-surgical routes.

[0102] Although the present disclosure and its advantages have been described in detail, the scope of the present invention is defined by the appended claims. Various changes, substitutions, and alterations may be made herein without departing from the spirit and scope of the present design. It is understood that the scope of the present application is limited to the The present invention is limited to certain embodiments of the process, machine, manufacture, composition of matter, means, methods and steps. As will be readily understood from this disclosure, those skilled in the art will appreciate that the present disclosure is not intended to Perform substantially the same function or achieve substantially the same result as the corresponding embodiment described in Any now existing or later developed process, machine, manufacture, composition of matter, method or technique that accomplishes the Any stage, method or step may be utilized in accordance with the present disclosure. The appended claims include within their scope such processes, machines, manufacture, compositions of matter, means, or methods. or steps.

[0103] (References) The following references and publications are referenced throughout this specification: To the extent that they provide exemplary procedural or other details supplementary to those set forth herein, specific are incorporated into the system. 1. Seth, P., et al., Overdose Deaths Involving Opioids, Cocaine, and Psych ostimulants-United States, 2015-2016. MMWR Morb Mortal Wkly Rep, 2 018. 67(12): p. 349-358. 2. Votaw, VR, et al., Perceived risk of heroin use among nonmedical prescr. iption opioid users. Addict Behav, 2017. 65: p. 218-223. 3. Al-Tayyib, A.A., S. Koester, and P. Riggs. Prescribing before injection drug use. Opioids: Comparative and public health implications. Addict Behav. 2017. 65: p. 224 -228. 4. Banerjee, G., et al., Nonmedical Use of Prescription Opioids in U.S. Military Veterans. Associated with heroin initiation: a prospective cohort study. Addiction. 2016. 111(11): p. 2021-2031. 5. Srivastava, A., M. Kahan, and M. Nader, Primary care management of opioi d use disorders: Abstinence, methadone, or buprenorphine-naloxone? Can Fam Phy sician, 2017. 63(3): p. 200-205. 6. Valenstein-Mah, H., et al., Underutilization of current clinical capacity to provide buprenorphine treatment for opioid use disorders within Veterans Hea lth Administration. Subst Abus, 2018: p. 1-3. 7. Ronquest, NA et al., Bupivacause and Treatment of Opioid Use Disorder in Privately and Publicly Insured Patients. The relationship between lenorphin adherence and relapse, health care utilization, and costs. Subst Abuse Rehabil, 20 18. 9: p. 59-78. 8. Leung, LS et al., Brain regions affected by general anesthesia. Prog Neurobiol, 2014. 122: p.24-44. 9. Vrana, KE, et al., Nonhuman primate parthenogenetic stem cells. Proc Na tl Acad Sci USA, 2003. 100 Appendix 1: p. 11911-6. 10. Sanchez-Pernaute, R. et al., Cellular Pathogenesis, Derived from Parthenogenetic Primate Embryonic Stem Cells (cyno-1) Long-term survival after transplantation of dopamine neurons. Stem Cells, 2005. 23(7): p. 914- twenty two. 11. Cibelli, J.B., K. Cunniff, and K.E. Vrana: Parthenogenetic derived embryonic stem cells. . Methods Enzymol, 2006. 418: p. 117-35. 12. Revazova, E.S., et al. Patient-specific stem cell lines derived from human parthenogenetic blastocysts. Stem. Cell Cloning, 2007. 9(3): p. 432-49. 13. de Fried, EP, et al., Human parthenogenetic blastocysts derived from n oninseminated cryopreserved human oocytes. Fertil Steril, 2008. 89(4): p. 943-7. 14. French, AJ, SH Wood, and AO Trounson, Human therapeutic cloning ( NTSC): Applications of research from mammalian reproductive cloning. Stem Cell Rev, 2006. 2( 4): p. 265-76. 15. Lin, G., et al., Highly purified oocytes derived from 1 pronucleated oocytes following an in vitro fertilization procedure. Homozygous parthenogenetic human embryonic stem cell lines. Cell Res, 2007. 17(12): p. 9 99-1007. 16. Rebazoba, E.S. et al. HLA homozygous stem cell lines derived from human parthenogenetic blastocysts. Stem Cell Cloning, 2008. 10(1): p. 11-24. 17. De Sousa, PA and I. Wilmut, Human parthenogenetic embryo stem cells: Appreciate what you have. Cell Stem Cell, 2007. 1(3): p. 243- 4. 18. Wun, IC and RE Dittman, Human somatic cell nuclear transfer. Chin J Ph ysiol, 2008. 51(4): p. 208-13. 19. Taupin, P. Parthenogenetic activation of human oocytes and parthenogenetic embryonic stem cells: US201 00233143. Expert Opin Ther Pat, 2011. 21(8): p. 1281-3. 20. Wei, Q. et al. Derivation of rhesus monkey parthenogenetic embryonic stem cell s and its microRNA signature. PLoS One, 2011. 6(9): pp e25052. . 21. Yabuuchi, A., H. Rehman, and K. Kim. Histocompatible parthenogenetic embryonic stem cells for regeneration. Potential sources of medical care. J Mamm Ova Res, 2012. 29(1): p. 17-21. 22. Daughtry, B. and S. Mitalipov. Concise review: Parthenogenetic stem cells for regenerative medicine. Cells: Genetic, epigenetic and developmental characteristics. Stem Cells Transl Med, 2014. 3(3): p.290-8. 23. Espejel, S., et al., Brief report: Parthenogenetic embryonic stem cells are a promising candidate for therapeutic liver repopulation. It is an effective cell source for stem cells. Stem Cells, 2014. 32(7): p. 1983-8. 24. Cervera, R. P. and M. Stojkovic, Human embryonic stem cell derivation and Nuclear transfer: Impact on regenerative medicine and drug discovery. Clin Pharmacol Ther, 2007. 82(3): p.310-5. 25. De Sousa, PA, et al., Clinically failed eggs as a source of normal hum an embryo stem cells. Stem Cell Research, 2009. 2(3): p. 188-97. 26. Takahashi, K. and Yamanaka, S. Regulation of mouse embryonic and adult fibroblast cell cultures by regulation factors. Induction of culture-derived pluripotent stem cells. Cell, 2006. 126(4): p. 663-76. 27. 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Claims

[Claim 1] A method of treating an addiction or substance abuse in a subject, comprising administering to a subject a fibroblast, a plurality of fibroblasts, and / or or a product derived therefrom, wherein the cells are / or a method for expressing the CD73 marker.