WO2007004469A1 - Procede de culture pour la differenciation d'un preadipocyte en adipocyte mature et criblage d'une substance susceptible d'influer sur le processus de differenciation du preadipocyte en adipocyte mature - Google Patents

Procede de culture pour la differenciation d'un preadipocyte en adipocyte mature et criblage d'une substance susceptible d'influer sur le processus de differenciation du preadipocyte en adipocyte mature Download PDF

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Publication number
WO2007004469A1
WO2007004469A1 PCT/JP2006/312793 JP2006312793W WO2007004469A1 WO 2007004469 A1 WO2007004469 A1 WO 2007004469A1 JP 2006312793 W JP2006312793 W JP 2006312793W WO 2007004469 A1 WO2007004469 A1 WO 2007004469A1
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Prior art keywords
preadipocytes
culture
differentiation
mature
mature adipocytes
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PCT/JP2006/312793
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English (en)
Japanese (ja)
Inventor
Akifumi Matsuyama
Yoshiki Sawa
Yuzuru Kanakura
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Osaka University
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Publication of WO2007004469A1 publication Critical patent/WO2007004469A1/fr

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    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/5005—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells
    • G01N33/5008—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics
    • G01N33/5044—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics involving specific cell types
    • 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
    • C12N5/0653—Adipocytes; Adipose tissue
    • 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
    • C12N2501/00—Active agents used in cell culture processes, e.g. differentation
    • C12N2501/999—Small molecules not provided for elsewhere

Definitions

  • the present invention relates to a culture method for differentiating preadipocytes into mature adipocytes, characterized by culturing preadipocytes in a suspended state, and mature adipocytes obtainable thereby,
  • a screening method for substances that affect the differentiation process from preadipocytes to mature adipocytes characterized by using a culture method, and the differentiation process from preadipocytes to mature adipocytes that can be obtained thereby Material, as well as kits for such screening.
  • arteriosclerotic diseases such as coronary artery disease and cerebrovascular disorder secondary to metabolic syndrome has increased. This is associated with almost all arteriosclerotic conditions, including metabolic syndrome is obesity, impaired glucose tolerance associated with insulin resistance, dyslipidemia, hypertension, and thrombus formation and inflammation with elevated PAI-1 and CRP, respectively. Because. Therefore, in order to prevent and treat these diseases, there is an urgent need for elucidation of the onset physiology of metabolic syndrome and drugs for treating it.
  • Adipocytes which are the main constituent cells of adipose tissue, are derived from adipose precursor cells, a fibroblast-like mesenchymal cell, and preadipocytes that encapsulate many lipid droplets, and then become unicellular Lipid droplets differentiate and mature into spherical mature adipocytes occupying most of the cells.
  • adipocytes produce many secretory factors (adipocytes), and it is known that these secretory factors are involved in the development of progeny Hi syndrome.
  • Preadipocytes produce secretory factors that suppress the development of metabolic syndrome such as adiponectin, but more mature mature adipocytes reduce the production of adiponectin and promote the development of metabolic syndrome such as TNF It is known to produce secretory factors. Therefore, by studying the differentiation process from preadipocytes to mature adipocytes and inhibiting the differentiation, although it is believed that the disease group can be prevented, a culture system that differentiates preadipocytes into mature fat cells has not been established so far, and substances having such inhibitory action are screened in vitro. It is impossible.
  • Non-Patent Document 1 Tchknoia T et al., Am J Physiol Endocrinol Metab 288: E267-E277 (2 005)
  • Non-Patent Document 2 Hemmrich K et al., Differentiation 73: 28-35 (2005)
  • the problem to be solved by the present invention affects the culture method of differentiating preadipocytes into spherical mature adipocytes encapsulating monocytic lipid droplets, and the differentiation process into mature adipocytes, such as preadipocyte force It is to provide a screening method for substances.
  • the present inventors have surprisingly found that the three-dimensional environment, that is, the two-dimensional environment such as on the culture dish, has not been obtained in a suspended state. It has been found that by culturing fat cells, mature adipocytes that are morphologically and physiologically very similar to fat cells in the animal body can be obtained, and the present invention has been completed.
  • the present invention provides:
  • a preadipocyte is characterized by culturing in a suspended state.
  • a culture method for differentiation into mature fat cells is characterized by culturing in a suspended state.
  • the cell is derived from a human, (1) to (3),
  • a mature adipocyte obtainable by the method according to (1) to (4),
  • a method for screening a substance that affects the differentiation process from preadipocytes to mature adipocytes characterized by using the culture method according to (1) or (2),
  • the cell is derived from a human, (6) The method according to,
  • a kit for screening a substance that affects the differentiation process from preadipocytes to mature adipocytes characterized by using the culture method according to (1) or (2),
  • the cell is derived from a human, (9) the kit according to
  • a method for culturing mature adipocytes and a method and kit for screening mature adipocytes obtainable thereby, and substances that affect the differentiation process from preadipocytes to mature adipocytes. Etc. are provided.
  • the present invention is excellent in that mature adipocytes can be obtained using human-derived preadipocytes, and that the obtained mature adipocytes are very similar to adipocytes present in vivo.
  • FIG. 1 is a micrograph of cells after culturing for 2 days in a state where preadipocytes are suspended.
  • FIG. 2 is a micrograph of mature adipocytes induced by culturing for 4 days in a state in which preadipocytes are suspended.
  • FIG. 3 is a micrograph of cells after preadipocytes have been cultured for 4 days under adhesion conditions.
  • FIG. 4 is a graph showing the influence of ECGC on the differentiation process from preadipocytes to mature adipocytes by the expression of adiponectin.
  • the white color indicates adiponectin expression in preadipocytes before the start of suspension culture, the black color indicates adiponectin expression after culturing the preadipocytes in the presence of ECGC for 4 days in the presence of ECGC.
  • FIG. 5 is a graph showing the effect of ECGC on the differentiation process from preadipocytes to mature adipocytes by the expression of PAI-1.
  • the white color indicates PAI-1 expression in preadipocytes before the start of suspension culture, the black color indicates PAI-1 expression in the absence of ECGC, and the hatched line indicates PAI-1 expression after 4 days in the presence of ECGC.
  • the present invention relates to a culture method for differentiating preadipocytes into mature adipocytes, characterized in that, in one embodiment, preadipocytes are cultured in a suspended state.
  • preadipocyte as used herein is defined as a cell that has differentiated from preadipocytes and encapsulates a large number of small lipid droplets.
  • mature adipocyte as used herein is defined as a globular cell that is similar to the adipocyte found in vivo and is predominantly composed of unicellular lipid droplets. Unless otherwise specified, other terms shall have the meanings commonly used in the field.
  • Preadipocytes can be obtained by means and methods known to those skilled in the art. For example, those isolated from visceral adipose tissue or subcutaneous adipose tissue, adipose precursor cells, or mesenchymal stem cells, or those induced to differentiate from stem cells such as ES cells may be used, but are not limited thereto.
  • the animal species from which the preadipocytes are derived is also not particularly limited, and preferably, for example, mammals including mice, rats, rabbits, dogs, cats, horses, horses, etc., more preferably humans. is there.
  • mammals including mice, rats, rabbits, dogs, cats, horses, horses, etc., more preferably humans. is there.
  • human preadipocytes it is possible to study the pathogenesis of metabolic syndrome, etc., or use self-matured adipocytes differentiated and proliferated in vitro for reconstruction after mastectomy in breast cancer patients It is also possible to do so.
  • the "suspended state" used in the present invention means placing cells in a released state by preventing or suppressing the adhesion or aggregation with a culture vessel or other cells.
  • Cell floating can be performed by various known means and methods.
  • cells may be placed in a floating state using a culture vessel or device that has been treated to prevent or inhibit cell adhesion, or made of a material that prevents or inhibits cell adhesion.
  • Examples of such a culture vessel or apparatus include a low-adhesion culture vessel such as a silicon-treated culture vessel (for example, a siliconized flask) or a low-adhesion culture dish.
  • the cells may be cultured in a suspended state by using a hanging drop culture method.
  • known means and methods may be used in combination as appropriate at the start of floating or in order to continue floating. Examples of methods that can be used are as follows. Alternatively, after culturing the cells in a temperature-responsive culture equipment for cell recovery (for example, CellSeed), the cells are detached by, for example, incubation at 20 ° C. for 30 minutes.
  • a temperature-responsive culture equipment for cell recovery for example, CellSeed
  • the present invention provides a mature fat cell obtainable by the above-described differentiation culture method.
  • Such mature adipocytes are very advantageous in that they have morphologically and physiologically similar properties to the fat cells in the animal body.
  • using mature adipocytes obtainable by the differentiation culture method of the present invention it is possible to conduct experiments closer to the living body at the in vitro port.
  • the present invention is characterized by administering to a subject mature adipocytes obtainable by the above-described culture method for differentiating preadipocytes into mature adipocytes.
  • the present invention relates to a method for treating or preventing diseases such as lipodystrophy.
  • treatment can be performed by performing the above method using preadipocytes derived from a subject with lipodystrophy and transplanting the resulting mature adipocytes into the subject.
  • the present invention uses a mature adipocyte obtainable by the above-described culture method for differentiating preadipocytes into mature adipocytes. It is related with the treatment method. This method can be applied to, for example, breast reconstruction and various cosmetic surgery. Preferably, problems such as side effects can be avoided by using mature adipocytes differentiated from autologous preadipocytes.
  • the present invention relates to a method for screening for a substance that affects the differentiation process from preadipocytes to mature adipocytes using the culture method described above.
  • to influence the differentiation process means to suppress or stop the differentiation from preadipocytes to mature adipocytes, or to promote a certain level. Specifically, decreasing the differentiation rate from preadipocytes to mature adipocytes, delaying the differentiation, incompletely or completely stopping the differentiation, reducing the number of mature adipocytes, Or, conversely, increasing the differentiation rate, increasing the number of mature adipocytes, and further immature differentiated mature adipocytes.
  • the screening method of the present invention is performed by adding a candidate substance to the differentiation culture system and examining differentiation from preadipocytes to mature adipocytes.
  • a candidate substance for example, the ability to include analogs, derivatives or mutants of ECGC (epicaro tectin gallate), which are known to suppress differentiation into preadipocyte force mature adipocytes. It is not limited to.
  • a means for examining differentiation for example, calculating the number of mature adipocytes by microscopic observation, observing the size, number, fat content, etc. of fat droplets by oil red staining, from preadipocytes Substances whose expression decreases or increases upon differentiation into mature adipocytes, such as aP2, CD36, adiponectin, resistin, GLUT4, TNF-s, PAI-1, etc. measured by quantitative PCR or immunoplot etc. Is mentioned.
  • the differentiation from preadipocytes to mature adipocytes in the differentiation culture system in the case where no candidate substance is added is also examined, and compared with the differentiation when the candidate substance is added. If the differentiation is suppressed in a candidate substance-added culture system, the candidate substance is considered to be a substance that suppresses the differentiation. If conversion is promoted, the candidate substance is considered to be a substance that promotes the differentiation.
  • a substance that suppresses differentiation into mature adipocytes using human-derived preadipocytes may be obesity, diabetes, or hypertension. It is considered to be a substance useful for the treatment or prevention of diseases such as hyperlipidemia, metabolic syndrome, arteriosclerotic disease, and ischemic heart disease. In addition, if a substance that promotes differentiation is found, it is considered to be a substance useful for the treatment or prevention of diseases such as malnutrition, lupus, and lipodystrophy. Since the screening method of the present invention uses, for example, the culture method of the present invention having the advantages as described above, it is an easy, efficient and economical method for finding such useful substances.
  • the use of the screening method of the present invention using patient-derived cells makes it possible to create drugs and cells for tailor-made treatment.
  • the present invention relates to a substance that can be obtained by the above screening method and that influences the pre-adipocyte force differentiation process into mature adipocytes.
  • a substance that promotes differentiation from preadipocytes into mature adipocytes may be used in the culture method of the present invention to increase the number of mature adipocytes obtained.
  • the present invention provides a kit for screening a substance that affects the differentiation process from preadipocytes to mature adipocytes, characterized by using the culture method described above.
  • the kit of the present invention may contain a container, an instrument, a medium, etc. for carrying out the suspension culture of the present invention. Usually, an instruction manual is attached to the kit.
  • means for facilitating screening and promoting efficiency 'method for example, oil red O staining solution for confirming differentiation into mature adipocytes, quantitative PCR Primers and / or probes, ELIS A, antibodies for immunoplots, etc. may be used.
  • Example 1 Differentiation of preadipocytes derived from preadipocytes into mature adipocytes Human adipose tissue-derived preadipocytes were added to a basic medium (DMEMZF-12 medium containing 10% FCS) at a concentration of 100,000 / mL. Then, 10 mL of this suspension was seeded on a 10 cm culture dish. The basal medium was changed the next day, and every 3 days thereafter, and adipose precursor cells were grown until confluent. Next, the basal medium was changed to a differentiation medium (DMEM / F-12 medium containing 10% FCS, 66 ⁇ m insulin, 250nM dexamethasone, 0.
  • DMEM / F-12 medium containing 10% FCS, 66 ⁇ m insulin, 250nM dexamethasone
  • Pre-adipocytes were obtained by culturing at 37 ° C for 3 days. The differentiation medium was aspirated and the preadipocytes were washed with PBS. Next, 1 mL of trypsin / EDTA solution was added and incubated at 37 ° C for 1 minute. 9 mL of basic medium was added and pipetted, and the resulting cell suspension was transferred to a 15 mL tube. The resulting cell pellet was centrifuged at 440 g for 2 minutes at 4 ° C. Medium) Suspended in 12 mL.
  • the prepared preadipocytes were suspended and cultured in a medium containing ECGC for 4 days.
  • RNA is extracted, and PCR (internal standard: GAPDH) is performed using the TaqMan [registered trademark] probe (Applied Bio Systems) with the strong RNA as a saddle type, and expression of adiponectin and PAI-1 was quantified. It was confirmed that the expression of adiponectin in mature adipocytes obtained by culturing in a medium containing ECGC was increased compared to cells cultured in a medium containing ECGC (Fig. 4). .
  • a culture method for differentiating preadipocytes into mature adipocytes encapsulating monocystic lipid droplets which are similar to adipocytes of visceral adipose tissue and subcutaneous adipose tissue, and substances that affect the differentiation process
  • a culture method for differentiating preadipocytes into mature adipocytes encapsulating monocystic lipid droplets which are similar to adipocytes of visceral adipose tissue and subcutaneous adipose tissue, and substances that affect the differentiation process
  • Can be used in the field of pharmaceuticals for example, in the field of development, manufacturing, or health foods for prophylactic, therapeutic or diagnostic agents for diseases such as deceitful syndromes.

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Abstract

La présente invention concerne un procédé de culture pour induire la différenciation d'un préadipocyte en un adipocyte mature, le procédé comprenant la culture du préadipocyte en flottaison ; un adipocyte mature produit grâce au procédé ; un procédé de criblage pour une substance susceptible d'influer sur le processus de différenciation d'un préadipocyte en un adipocyte mature, caractérisé par l'utilisation du procédé de culture ; une substance susceptible d'influer sur le processus de différenciation d'un préadipocyte en un adipocyte mature, qui est obtenue grâce au procédé de criblage ; un kit pour le criblage ; et autres.
PCT/JP2006/312793 2005-07-04 2006-06-27 Procede de culture pour la differenciation d'un preadipocyte en adipocyte mature et criblage d'une substance susceptible d'influer sur le processus de differenciation du preadipocyte en adipocyte mature WO2007004469A1 (fr)

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JP2005-195268 2005-07-04
JP2005195268 2005-07-04

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Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP6021802B2 (ja) * 2011-03-16 2016-11-09 株式会社クラレ 培養方法及び薬物スクリーニング方法
WO2019098310A1 (fr) * 2017-11-16 2019-05-23 日産化学株式会社 Méthode d'induction de la différenciation en et de production d'adipocytes beiges et blancs

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2000083656A (ja) * 1998-09-09 2000-03-28 Meiji Milk Prod Co Ltd 前駆脂肪細胞株
JP2000217576A (ja) * 1999-02-02 2000-08-08 Herikkusu Kenkyusho:Kk 脂肪細胞への分化を誘導する方法、並びに脂肪細胞への分化を制御する化合物およびそのスクリーニング方法
JP2003304866A (ja) * 2002-04-17 2003-10-28 National Institute Of Advanced Industrial & Technology 三次元凝集塊形成による細胞の分化制御

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2000083656A (ja) * 1998-09-09 2000-03-28 Meiji Milk Prod Co Ltd 前駆脂肪細胞株
JP2000217576A (ja) * 1999-02-02 2000-08-08 Herikkusu Kenkyusho:Kk 脂肪細胞への分化を誘導する方法、並びに脂肪細胞への分化を制御する化合物およびそのスクリーニング方法
JP2003304866A (ja) * 2002-04-17 2003-10-28 National Institute Of Advanced Industrial & Technology 三次元凝集塊形成による細胞の分化制御

Non-Patent Citations (5)

* Cited by examiner, † Cited by third party
Title
HEMMRICH K. ET AL.: "Optimization of the differentiation of human preadipocytes in vitro", DIFFERENTIATION, vol. 73, no. 1, February 2005 (2005-02-01), pages 28 - 35, XP003005177 *
KUROSAWA H. ET AL.: "A simple method for forming embryoid body from mouse embryonic stem cells", J. BIOSCI. BIOENG., vol. 96, no. 4, 2003, pages 409 - 411, XP004973487 *
PAIRAULT J. ET AL.: "A study of the adipose conversion of suspended 3T3 cells by using glycerophosphate dehydrogenase as differentiation marker", PROC. NATL. ACAD. SCI. USA, vol. 76, no. 10, 1979, pages 5138 - 5142, XP003005176 *
PETTERSSON P. ET AL.: "Cells in human adipose tissue developing into adipocytes", ACTA MED. SCAND., vol. 215, no. 5, 1984, pages 447 - 451, XP003007230 *
TCHKONIA T. ET AL.: "Abudance of two human preadipocyte subtypes with distinct capacities for replication, adipogenesis, and apoptosis varies among fat depots", AM. J. PHYSIOL. ENDOCRINOL. METAB., vol. 288, no. 1, January 2005 (2005-01-01), pages E267 - E277, XP003005178 *

Cited By (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP6021802B2 (ja) * 2011-03-16 2016-11-09 株式会社クラレ 培養方法及び薬物スクリーニング方法
WO2019098310A1 (fr) * 2017-11-16 2019-05-23 日産化学株式会社 Méthode d'induction de la différenciation en et de production d'adipocytes beiges et blancs
JPWO2019098310A1 (ja) * 2017-11-16 2020-11-19 日産化学株式会社 ベージュおよび白色脂肪細胞への分化誘導および生産法
EP3708654A4 (fr) * 2017-11-16 2020-12-23 Nissan Chemical Corporation Méthode d'induction de la différenciation en et de production d'adipocytes beiges et blancs
US11530387B2 (en) 2017-11-16 2022-12-20 Nissan Chemical Corporation Method for inducing differentiation into and producing beige and white adipocytes
JP7322707B2 (ja) 2017-11-16 2023-08-08 日産化学株式会社 ベージュおよび白色脂肪細胞への分化誘導および生産法
TWI820055B (zh) * 2017-11-16 2023-11-01 日商日產化學股份有限公司 成為米色及白色脂肪細胞之分化誘導及生產方法

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