WO2013168402A1 - Suspension de cellules de mammifère contenant du dextrane destinée à la prévention de la formation d'une embolie pulmonaire - Google Patents

Suspension de cellules de mammifère contenant du dextrane destinée à la prévention de la formation d'une embolie pulmonaire Download PDF

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WO2013168402A1
WO2013168402A1 PCT/JP2013/002924 JP2013002924W WO2013168402A1 WO 2013168402 A1 WO2013168402 A1 WO 2013168402A1 JP 2013002924 W JP2013002924 W JP 2013002924W WO 2013168402 A1 WO2013168402 A1 WO 2013168402A1
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mammalian
cells
dextran
stem cells
cell
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Japanese (ja)
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圭樹 和田
土居　雅子
健志 菊地
英司 小林
工 寺谷
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株式会社大塚製薬工場
学校法人自治医科大学
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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
    • A61K35/48—Reproductive organs
    • A61K35/54—Ovaries; Ova; Ovules; Embryos; Foetal cells; Germ cells
    • A61K35/545—Embryonic stem cells; Pluripotent stem cells; Induced pluripotent stem cells; Uncharacterised stem cells
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/30—Macromolecular organic or inorganic compounds, e.g. inorganic polyphosphates
    • A61K47/36—Polysaccharides; Derivatives thereof, e.g. gums, starch, alginate, dextrin, hyaluronic acid, chitosan, inulin, agar or pectin
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/0012—Galenical forms characterised by the site of application
    • A61K9/0019—Injectable compositions; Intramuscular, intravenous, arterial, subcutaneous administration; Compositions to be administered through the skin in an invasive manner
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K9/00—Medicinal preparations characterised by special physical form
    • A61K9/10—Dispersions; Emulsions
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P11/00—Drugs for disorders of the respiratory system
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
    • C12N5/06—Animal cells or tissues; Human cells or tissues
    • C12N5/0602—Vertebrate cells
    • C12N5/0652—Cells of skeletal and connective tissues; Mesenchyme
    • C12N5/0662—Stem cells
    • C12N5/0667—Adipose-derived stem cells [ADSC]; Adipose stromal stem cells

Definitions

  • the present invention relates to a mammalian cell suspension for preventing pulmonary embolism when a mammalian cell is administered via a blood vessel, a dextran or a derivative thereof, comprising the mammalian cell and dextran or a derivative thereof or a salt thereof.
  • the present invention relates to a prophylactic agent for pulmonary embolism when a mammalian cell is administered via a blood vessel, containing a salt thereof as an active ingredient.
  • Regenerative medicine using stem cells aims to restore the functions of cells and tissues damaged by various diseases by utilizing the self-replicating ability and pluripotency of stem cells and factors secreted by stem cells. Medical care. When bone marrow transplantation is performed in patients with intractable blood diseases such as leukemia and aplastic anemia, hematopoietic stem cells are engrafted in the patient's body, and hematopoietic ability can be maintained almost throughout the life.
  • Non-Patent Documents 1 to 3 are tissue stem cells that are characterized by hematopoietic stem cells. identifying stem cells in the central nerve, peripheral nerve, bone marrow, small intestine, etc., and transplanting tissue stem cells for traumatic or tissue degenerative diseases Treatment has begun to be practiced.
  • stem cell transplantation In order for mammalian stem cells to be transplanted efficiently to the target injury site (affected site), it is considered important to optimize the stem cell transplantation method (transplant route). To date, there are mainly four types of stem cell transplantation methods: stereotactic [direct] transplantation, intrathecal [intracerebral spinal cord] transplantation, transvenous transplantation, and transarterial transplantation. The law is known. Among these, stereotactic transplantation is a method in which stem cells are directly transferred to the affected area. When stereotactic transplantation is used, the invasion is somewhat large, but since it is administered directly to the affected area, there are many engrafted donor stem cells, and therefore the number of stem cells to be administered can be reduced.
  • the intrathecal transplantation method is a method of transferring stem cells into the medullary canal by ventricular puncture.
  • Intrathecal transplantation is a method that has been studied mainly to treat brain diseases such as cerebral infarction, cerebral contusion, and spinal cord injury, but there is a potential risk that ventricular puncture may cause new brain damage. Many problems still remain in clinical application.
  • Transvenous transplantation and transarterial transplantation are methods in which cells such as stem cells are transferred into veins and arteries, respectively (administration via blood vessels).
  • the number of donor hepatocytes to engraft is less than when using stereotaxic transplantation, but it is minimally invasive and secretes cells such as stem cells and stem cells throughout the body. Factors can be circulated.
  • a method of transplanting mesenchymal stem cells (MSC) intravenously for cerebral infarction disease or mononuclear for cerebral infarction disease As a method of administration via blood vessels, which is practiced in clinical practice, a method of transplanting mesenchymal stem cells (MSC) intravenously for cerebral infarction disease or mononuclear for cerebral infarction disease
  • MSC mesenchymal stem cells
  • a method of transplanting spheres intravenously, a method of transplanting islet cells intravenously in type I diabetic patients, and the like are known.
  • stem cells As a risk when transplanting cells such as stem cells using the vascular administration method, cells that have been transarterally or intravenously administered are clogged with capillaries of the pulmonary artery (pulmonary embolism) when passing through the lung, As a result, it has been pointed out that there is a risk of pulmonary function or cardiac function (pulmonary embolism) and possibly death.
  • stem cells are transplanted while monitoring peripheral oxygen partial pressure during stem cell administration with a pulse oximeter in clinical practice.
  • dextran is a kind of polysaccharide consisting of glucose and is widely used as a thickener, moisturizer, etc. in the fields of pharmaceuticals and cosmetics.
  • dextran is a kind of polysaccharide consisting of glucose and is widely used as a thickener, moisturizer, etc. in the fields of pharmaceuticals and cosmetics.
  • cells such as stem cells are suspended in a solution containing dextran and the cell suspension is administered via a blood vessel, the risk of pulmonary embolism due to cells is reduced and pulmonary embolism is prevented. It was unknown.
  • An object of the present invention is to provide a mammalian cell suspension that can prevent pulmonary embolism when a mammalian cell such as a mammalian stem cell is administered via a blood vessel, or a mammalian cell suspension that is administered via a blood vessel.
  • the object is to provide a preventive agent for pulmonary embolism.
  • the present inventors have found that suspension of mammalian stem cells in a solution containing dextran suppresses aggregation of mammalian stem cells and decreases the survival rate (Japanese Patent Application No. 2010-251273). ).
  • the aggregate of stem cells is blocking capillaries of pulmonary arteries, but the size of stem cells, for example, mesenchymal stem cells (MSCs)
  • MSCs mesenchymal stem cells
  • the pulmonary artery capillaries have an inner diameter of 10 ⁇ m, whereas the MSC is larger than the pulmonary artery capillaries, whereas the pulmonary embolization by stem cells is simply inhibited by cell aggregation. It was thought enough not to interfere with.
  • the present inventors suspend adipose tissue-derived mesenchymal stem cells (AT-MSC) in a lactate Ringer solution containing dextran, and suspend the AT-MSC suspension dextran-containing lactate Ringer solution in pigs.
  • AT-MSC adipose tissue-derived mesenchymal stem cells
  • the present invention includes (1) a mammalian cell suspension for preventing pulmonary embolism when a mammalian cell is administered via a blood vessel, including the mammalian cell and dextran or a derivative thereof or a salt thereof, (2) the mammalian cell is a mammalian stem cell, the mammalian cell suspension according to (1) above, or (3) the mammalian stem cell is a mammalian mesenchymal stem cell or a mammalian pluripotent stem cell.
  • the mammalian cell suspension according to any of (1) to (4) above, wherein the concentration of the suspension or (5) dextran or a derivative thereof or a salt thereof is in the range of 0.1 to 20%.
  • the present invention also relates to (6) an agent for preventing pulmonary embolism when a mammalian cell is administered via a blood vessel, comprising dextran or a derivative thereof or a salt thereof as an active ingredient, and (7) the mammalian cell is a mammal.
  • the prophylactic agent according to (6) which is a stem cell
  • the prophylactic agent according to (7) wherein the mammalian stem cell is a mammalian mesenchymal stem cell or a mammalian pluripotent stem cell.
  • the present invention relates to (9) use of dextran or a derivative thereof or a salt thereof for preparing a preventive agent for pulmonary embolism when a mammalian cell is administered via a blood vessel, or (10) a mammalian cell is sucked.
  • the risk of pulmonary embolism can be reduced when mammalian cells such as mammalian stem cells are administered via blood vessels, and the risk of developing pulmonary embolism can be reduced.
  • Pulmonary artery pressure (PA [PA [] in pigs each administered two types of AT-MSC suspensions (AT-MSC suspension physiological saline [... ⁇ ...] and AT-MSC suspension dextran-containing lactated Ringer's solution [- ⁇ -]). It is a figure which shows the result of having measured mmHg]).
  • the vertical axis represents pulmonary artery pressure.
  • the pulmonary artery pressure is expressed as a relative value when divided by the value measured before administration of AT-MSC suspension (0 minutes).
  • the horizontal axis indicates the time after the start of administration of the AT-MSC suspension.
  • Blood oxygen partial pressure (%) in pigs each administered two types of AT-MSC suspensions (AT-MSC suspension physiological saline [ ⁇ ] and AT-MSC suspension dextran-containing lactated Ringer's solution [ ⁇ ]) It is a figure which shows the measurement result.
  • the vertical axis represents blood oxygen partial pressure.
  • the blood oxygen partial pressure is expressed as a relative value when the value measured before administration of the AT-MSC suspension (0 minutes) is taken as 100%.
  • the horizontal axis indicates the time after the start of administration of the AT-MSC suspension.
  • Mammalian cell suspensions for preventing pulmonary embolism when administering mammalian cells such as mammalian stem cells of the present invention via blood vessels include mammalian cells such as mammalian stem cells and dextran or derivatives thereof, or The suspension is not particularly limited as long as it is a suspension containing a salt thereof (hereinafter referred to as dextran), and a preventive agent for pulmonary embolism when a mammalian cell such as the mammalian stem cell of the present invention is administered via a blood vessel. Is not particularly limited as long as it is a composition containing dextran as an active ingredient.
  • mammals examples include rodents such as mice, rats, hamsters, and guinea pigs, rabbits such as rabbits, pigs, cows, and goats. Horses, sheep, ungulates, dogs, cats, etc., humans, monkeys, rhesus monkeys, cynomolgus monkeys, marmosets, orangutans, chimpanzees, etc.
  • Examples of the algae, among others, mouse, pig, and human can be preferably exemplified, and as the mammalian cell, in addition to mammalian stem cells administered via revascularization to regenerative medicine, etc.
  • stem cell means an immature cell having self-renewal ability and differentiation / proliferation ability.
  • Stem cells include subpopulations such as pluripotent stem cells (pluripotent stem ce11), multipotent stem cells (multipotent stem ce11), unipotent stem cells (unipotent stem ce11), etc., depending on their differentiation ability.
  • pluripotent stem cell means a cell that cannot be an individual by itself, but has the ability to differentiate into all tissues and cells constituting a living body.
  • a multipotent stem cell means a cell having the ability to differentiate into multiple types of tissues and cells, although not all types.
  • a unipotent stem cell means a cell having the ability to differentiate into a specific tissue or cell.
  • pluripotent stem cells examples include embryonic stem cells (ES cells), EG cells, iPS cells, and the like.
  • ES cells can be produced by culturing the inner cell mass on feeder cells or in a medium containing LIF. Methods for producing ES cells are described in, for example, WO96 / 22362, WO02 / 101057, US5,843,780, US6,200,806, US6,280,718 and the like.
  • EG cells can be produced by culturing primordial germ cells in a medium containing mSCF, LIF and bFGF (Ce11, 70: 841-847, 1992).
  • iPS cells are produced by introducing reprogramming factors such as Oct3 / 4, Sox2 and Klf4 (c-Myc or n-Myc as necessary) into somatic cells (eg, fibroblasts, skin cells, etc.).
  • somatic cells eg, fibroblasts, skin cells, etc.
  • Stem cells established by culturing early embryos produced by nuclear transfer of somatic cell nuclei are also preferred as pluripotent stem cells (Nature, 385, 810 (1997); Science, 280, 1256 (1998); Nature Biotechnology, 17, 456 (1999); Nature, 394, 369 (1998); Nature s Genetics, 22, 127 (1999); Proc. Nat1. Acad. Sci. USA, 96, 14984 (1999)), Rideout III et al. (Nature Genetics, 24, 109 (2000) )).
  • Multipotent stem cells include mesenchymal stem cells that can differentiate into cells such as adipocytes, bone cells, chondrocytes, adipocytes, hematopoietic stem cells that can differentiate into blood cells such as leukocytes, erythrocytes, and platelets, neurons, Examples include neural stem cells that can differentiate into cells such as astrocytes and oligodendrocytes, somatic stem cells such as bone marrow stem cells and germ stem cells.
  • the multipotent stem cell is preferably a mesenchymal stem cell.
  • mesenchymal stem cell is meant a stem cell that can differentiate into all or some of osteoblasts, chondroblasts and lipoblasts.
  • Functional stem cells can be isolated from a living body by a method known per se.
  • mesenchymal stem cells can be collected by known general methods from mammalian bone marrow, adipose tissue, peripheral blood, umbilical cord blood and the like.
  • human mesenchymal stem cells can be isolated by culturing and passage of hematopoietic stem cells after bone marrow puncture (Journal of Autoimmunity, 30 (2008) 163-171).
  • Multipotent stem cells can also be obtained by culturing the pluripotent stem cells under appropriate induction conditions.
  • Examples of cells such as stem cells contained in the mammalian cell suspension of the present invention include adherent cells.
  • Adherent cells are likely to aggregate in the suspension, but since the suspension of the present invention contains dextrans, this aggregation can be effectively suppressed.
  • the “adhesive” cell means an anchorage-dependent cell that can survive, proliferate, and produce a substance by adhering to the scaffold.
  • adherent stem cells include pluripotent stem cells, mesenchymal stem cells, nervous system stem cells, bone marrow stem cells, and reproductive stem cells.
  • the adherent stem cells are preferably mesenchymal stem cells or pluripotent stem cells.
  • Mammalian cells may be isolated from the living body or subcultured in vitro. Moreover, it is preferable that the mammalian cell (population) contained in the mammalian cell suspension of the present invention is isolated or purified.
  • isolation or purification means that an operation for removing components other than the target component has been performed.
  • the purity of the isolated or purified mammalian cells is usually 30% or more, preferably 50% or more, more preferably 70% or more, More preferably, it is 90% or more (for example, 100%).
  • the mammalian cells (population) contained in the suspension of the present invention include mammalian cells in a single cell (single cell) state.
  • the “single cell state” means that the cells do not gather together to form a lump (that is, they are not aggregated).
  • Mammalian cells in a single cell state can be prepared by enzymatic treatment of mammalian cells cultured in vitro with trypsin / EDTA or the like.
  • the ratio of mammalian cells in a single cell state contained in mammalian cells is usually 70% or more, preferably 90% or more, more preferably 95% or more, and further preferably 99% or more (for example, 100%). is there.
  • the percentage of cells in a single cell state is determined by dispersing mammalian cells in PBS and observing them under a microscope, and examining the presence or absence of aggregation of a plurality of randomly selected cells (eg, 1000). Can be determined.
  • the mammalian cells are preferably floating.
  • “floating” means that cells are held in the suspension without contacting the inner wall of the container containing the suspension.
  • the dextran in the dextran used in the present invention is not particularly limited as long as it is a polysaccharide (C 6 H 10 O 5 ) n consisting of D-glucose and having an ⁇ 1 ⁇ 6 bond as the main chain.
  • These dextrans can be produced by any known method such as chemical synthesis, production by microorganisms, production by enzymes, etc., but commercially available products can also be used.
  • commercially available products such as low molecular weight dextran L injection (manufactured by Otsuka Pharmaceutical Factory) and dextran 70 (manufactured by Tokyo Chemical Industry Co., Ltd.) can be mentioned.
  • Dextran derivatives in dextrans used in the present invention include dextran sulfate, carboxylated dextran, diethylaminoethyl (DEAE) -dextran and the like.
  • Examples of the salt of dextran or a derivative thereof used in the present invention include hydrochloride, hydrobromide, hydroiodide, phosphate, nitrate, sulfate, acetate, propionate, toluene Sulfonate, succinate, oxalate, lactate, tartrate, glycolate, methanesulfonate, butyrate, valerate, citrate, fumarate, maleate, malate, etc.
  • Acid addition salts, metal salts such as sodium salts, potassium salts and calcium salts, ammonium salts and alkylammonium salts. These salts are used as a solution at the time of use, and the action is preferably the same as that of dextran. These salts may form a hydrate or a solvate, and can be used alone or in combination of two or more.
  • the concentration of dextran applied to the mammalian cell suspension or the pulmonary embolization prevention agent of the present invention may be any concentration that can prevent pulmonary embolization by cells such as stem cells.
  • the concentration may be appropriately selected depending on the concentration, but a concentration sufficient to suppress aggregation of mammalian cells and a decrease in survival rate is preferable.
  • the concentration of dextran applied to the mammalian cell suspension or the pulmonary embolism prevention agent of the present invention is usually 0.1% or more, preferably 0.5% or more, more preferably 1.0%. That's it.
  • the concentration of dextran in the suspension is, for example, 20% or less, preferably 10% or less, more preferably 7% or less, and even more preferably 5%. It is as follows. Therefore, the concentration of dextran in the suspension is, for example, 0.1 to 20%, preferably 0.5 to 10%, more preferably 1.0 to 7.0%, and still more preferably 1.0 to 5.0%.
  • pulmonary embolization when administered via a blood vessel means transmural or intravenous implantation (administration) of mammalian cells in the peripheral arterial system of the lung, for example, one or more capillaries of the pulmonary artery. It means forming a state in which blood vessels are blocked.
  • the state in which the transplanted mammalian cells “clog” the peripheral arterial system of the lung does not necessarily mean that the cell has stopped in the peripheral arterial system of the lung, and obstructs the blood flow at that location. It is enough if you do. In this case, the blood flow does not need to be completely stopped, and it is sufficient that the blood flow is reduced to such a degree that the lung function is reduced in a broad sense.
  • the “pulmonary function” primarily means the uptake of oxygen from the atmosphere into the blood, but also includes the discharge of carbon dioxide and other gases.
  • pulmonary embolization occurs due to cells, blood flow in capillaries of the pulmonary artery decreases, causing tachypnea (increased respiratory rate) and tachycardia (increased heart rate), and blood pressure such as pulmonary artery pressure increases Or, the partial pressure of oxygen in the blood decreases. For this reason, by preventing the formation of pulmonary embolism by cell transplantation, it is possible to reduce risks such as tachypnea, tachycardia, blood pressure increase and blood oxygen partial pressure decrease due to pulmonary embolism.
  • pulmonary embolism when pulmonary embolism is formed by cell transplantation, depending on the case, the lung function may decrease or the cardiac function (the function of moving blood into the artery or the function of moving the blood out of the vein) may decrease.
  • the lung function may decrease or the cardiac function (the function of moving blood into the artery or the function of moving the blood out of the vein) may decrease.
  • pulmonary embolization in the present invention includes cases where mammalian cells form a state in which other blood vessels are blocked in addition to the peripheral arterial system of the lung.
  • mammalian cells are usually suspended in a physiological aqueous solution containing dextran as an active ingredient.
  • the preventive agent for pulmonary embolization according to the present invention is roughly classified into a liquid type and a non-liquid type.
  • the liquid type preventive agent is usually constituted as a physiological aqueous solution containing dextran as an active ingredient, and the mammalian cell suspension of the present invention can be prepared by suspending mammalian cells in such a liquid type preventive agent. it can.
  • the non-liquid type preventive agent is usually configured as a dextran-containing material such as powder added to a physiological aqueous solution in which mammalian cells are suspended, and the mammalian cell suspends the non-liquid type preventive agent.
  • a dextran-containing material such as powder added to a physiological aqueous solution in which mammalian cells are suspended
  • the mammalian cell suspends the non-liquid type preventive agent When added to a turbid physiological aqueous solution, the mammalian cell suspension of the present invention can be prepared.
  • pulmonary embolism is prevented in cell transplantation of stem cells or the like in which a mammalian cell suspension containing mammalian cells and dextrans is administered via blood vessels.
  • a mammalian cell suspension containing mammalian cells and dextrans is administered via blood vessels.
  • use of dextran for preparing a prophylactic agent for pulmonary embolism when a mammalian cell is administered via a blood vessel can be mentioned.
  • physiological aqueous solution examples include physiological saline, phosphate buffered physiological saline, Tris buffered physiological saline, HEPES buffered physiological saline, Ringer's solution (lactate Ringer's solution, acetated Ringer's solution, bicarbonated Ringer's solution, etc.), 5 % Isotonic aqueous solution such as aqueous solution of glucose solution, mammalian culture medium, isotonic agents (glucose, D-sorbitol, D-mannitol, lactose, sodium chloride, etc.), among them Ringer's solution
  • lactate Ringer solution, acetate Ringer solution or bicarbonate Ringer solution is more preferable, and lactate Ringer solution is more preferable.
  • “isotonic” means that the osmotic pressure is in the range of 250 to 380 mOsm / l.
  • the physiological aqueous solution further contains a stabilizer (eg, human serum albumin, polyethylene glycol, etc.), a buffer (eg, phosphate buffer, sodium acetate buffer), a chelating agent (eg, EDTA, EGTA, citric acid, salicylate). ), Solubilizers, preservatives, antioxidants, and the like.
  • the present inventors can use trehalose, which has been shown to have a preventive action against pulmonary embolism, in combination with the dextran.
  • Cells such as stem cells contained in the cell suspension of the present invention are present in a suspended state, and normally suspended cells are likely to aggregate, but due to the effects of the dextrans of the present invention, cell aggregation is suppressed, The state of a single cell can be maintained for a long time.
  • Suspension of mammalian cells in a physiological aqueous solution containing dextran can be carried out by methods well known in the art such as pipetting and tapping.
  • the temperature of the mammalian cell suspension of the present invention is usually in the range of 0 to 37 ° C, preferably 0 to 25 ° C.
  • the density of the mammalian cells in the suspension of the present invention may be any density that can prevent the formation of pulmonary embolism by cells such as stem cells, but the effect of suppressing the aggregation of mammalian cells and the decrease in the survival rate due to dextrans Is more preferable, and is usually in the range of 10 3 to 10 10 pieces / ml.
  • the aggregation of mammalian cells is suppressed by dextrans, so that cell aggregates are clogged in the cannula by performing cell transplantation using this. Risk can be reduced.
  • dextrans suppress a decrease in the survival rate of mammalian cells in the suspension. Cell transplantation can be performed with cells, and an improvement in therapeutic effect can be expected.
  • the preventive agent for mammalian cell suspension or pulmonary embolism formation of the present invention can be produced as a mammalian cell suspension preparation or a preventive agent for pulmonary embolism formation by storing it in an appropriate sterile container.
  • containers include plastic bags such as bottles, vials, syringes, infusion bags, and test tubes. These containers may be formed from a variety of materials such as glass or plastic.
  • a cannula and / or injection needle can be connected to the container for mammalian cell suspension preparation so that the mammalian cell suspension of the present invention in the container can be instilled into a patient. .
  • Lactated Ringer's solution (dextran 40 added) (Savizol infusion) (Otsuka Pharmaceutical Factory, Tokushima, Japan) containing dextran (dextran 40) having a weight average molecular weight of 40,000 at a concentration of 30 mg / ml, and weight average molecular weight Contains 6.5% (65 mg / ml) dextran by adding an equal amount of low molecular weight dextran L injection (Otsuka Pharmaceutical Factory, Tokushima, Japan) containing 40000 dextran (dextran 40) at a concentration of 100 mg / ml After preparing the lactated Ringer's solution and adding it to the infusion bag, suck the appropriate amount of solution with a syringe, add the washed PBS (-) to the removed cells, and resuspend (30G needle [Nipro, Tokyo, Japan] Using a 5 mL syringe [Nipro, Tokyo, Japan], sucking and
  • an physiological saline solution (Otsuka Pharmaceutical Factory, Tokushima, Japan) was used as a cell suspension to prepare an AT-MSC suspension physiological saline solution (8.4 ⁇ 10 5 cells / mL).
  • AT-MSC suspension physiological saline solution 8.4 ⁇ 10 5 cells / mL.
  • AT-MSC when AT-MSC is transplanted using a dextran-free solution, AT-MSC accumulates in the lungs, particularly in the pulmonary capillaries, thereby forming pulmonary emboli, reducing blood flow in the lungs and increasing pulmonary artery pressure.
  • MSC when MSC is transplanted using a solution containing dextran, AT-MSC accumulation in the lung (formation of pulmonary embolism) is suppressed, blood flow in the lung does not decrease, and pulmonary artery pressure does not increase. .
  • Lactated Ringer's solution (dextran 40 added) (Savizol infusion) (Otsuka Pharmaceutical Factory, Tokushima, Japan) containing dextran (dextran 40) having a weight average molecular weight of 40,000 at a concentration of 30 mg / ml, and weight average molecular weight Contains 6.5% (65 mg / ml) dextran by adding an equal amount of low molecular weight dextran L injection (Otsuka Pharmaceutical Factory, Tokushima, Japan) containing 40000 dextran (dextran 40) at a concentration of 100 mg / ml After preparing 100 mL of lactated Ringer's solution and adding it to the infusion bag, suck out an appropriate amount of solution with a syringe, add the washed PBS (-) to the removed cells, and resuspend (30G needle [Nipro, Tokyo, Japan] And 5 mL syringe [Nipro, Tokyo, Japan] 3
  • cells were collected from 120 of the 240 culture dishes prepared through the same operation, and physiological saline (Otsuka Pharmaceutical Factory, Tokushima, Japan) was used as the cell suspension.
  • Suspension physiological saline (1.7 ⁇ 10 6 cells / mL) was prepared.
  • Each infusion bag is filled with a solution in which 100 mL of cells are suspended.
  • [5] Return two AT-MSC suspensions (AT-MSC suspension physiological saline and AT-MSC suspension dextran-containing lactated Ringer's solution) to separate infusion bags, and shake the infusion bag multiple times to Was stirred and prepared for cell infusion (transplantation).
  • FIG. 2 and Table 1 show the values obtained by measuring the blood oxygen partial pressure every 5 minutes from the start of administration (transplantation) as the relative values normalized by the blood oxygen partial pressure at the start of administration.
  • the blood oxygen partial pressure began to decrease after the start of administration, and the blood oxygen partial pressure decreased by about 19% at 15 minutes after administration compared to before administration.
  • FIG. 2, Table 1 At 25 minutes after the start of cell administration, the blood oxygen partial pressure decreased by about 42% compared to before administration.
  • pulmonary embolism can be prevented and the risk of developing pulmonary embolism can be reduced. It is useful in.

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Abstract

Le problème décrit par la présente invention est de fournir : une suspension de cellules de mammifère capable de prévenir la formation d'embolies pulmonaires lorsque des cellules souches de mammifère ou d'autres cellules de mammifère sont administrées par voie intraveineuse ; ou un agent prophylactique contre la formation d'embolies pulmonaires lorsque des cellules de mammifère sont administrées par voie intraveineuse.　Cette invention est caractérisée en ce que la suspension de cellules souches de mammifère ou d'autres cellules de mammifère dans une solution aqueuse physiologique comprend comme constituant efficace du dextrane, un de ses dérivés, ou un sel du dextrane ou d'un de ses dérivés. L'invention concerne également la préparation de la suspension de cellules de mammifère destinée à la prévention de la formation d'embolies pulmonaires lorsque des cellules de mammifère sont administrées par voie intraveineuse, ladite suspension contenant comme constituant efficace des cellules de mammifère et du dextrane, un de ses dérivés, ou un sel du dextrane ou d'un de ses dérivés. En plus des cellules souches de mammifère, des îlots de Langerhans, des cellules dendritiques, des cellules tueuses naturelles, des lymphocytes T alpha-beta, des lymphocytes T gamma-delta, et des lymphocytes T cytotoxiques, etc., peuvent être utilisés comme cellules de mammifère. Les cellules souches mésenchymateuses de mammifère et les cellules souches pluripotentes de mammifère sont des exemples appropriés de cellules souches de mammifère.
PCT/JP2013/002924 2012-05-08 2013-05-02 Suspension de cellules de mammifère contenant du dextrane destinée à la prévention de la formation d'une embolie pulmonaire WO2013168402A1 (fr)

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JP2012106909 2012-05-08
JP2012-106909 2012-05-08
JP2012230496A JP2013252126A (ja) 2012-05-08 2012-10-18 デキストラン含有肺塞栓形成予防用哺乳動物細胞懸濁液
JP2012-230496 2012-10-18

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WO2018123628A1 (fr) * 2016-12-28 2018-07-05 ロート製薬株式会社 Composition pharmaceutique de cellules, kit pour traitement de maladie, et solution pour suspension de cellules
JP6594578B1 (ja) * 2018-04-25 2019-10-23 セルトラスト・アニマル・セラピューティクス株式会社 細胞の保存方法および細胞懸濁液

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JPWO2016068228A1 (ja) * 2014-10-29 2017-04-27 株式会社高研 薬剤徐放担体及び薬剤徐放方法

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