JP2010220525A - Apparatus and method for separating stem cell - Google Patents

Apparatus and method for separating stem cell Download PDF

Info

Publication number
JP2010220525A
JP2010220525A JP2009070562A JP2009070562A JP2010220525A JP 2010220525 A JP2010220525 A JP 2010220525A JP 2009070562 A JP2009070562 A JP 2009070562A JP 2009070562 A JP2009070562 A JP 2009070562A JP 2010220525 A JP2010220525 A JP 2010220525A
Authority
JP
Japan
Prior art keywords
filter
space
stem cells
stem cell
container
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Withdrawn
Application number
JP2009070562A
Other languages
Japanese (ja)
Inventor
Koka Rin
孔華 林
Takahiro Ono
隆弘 大野
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Olympus Corp
Original Assignee
Olympus Corp
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Olympus Corp filed Critical Olympus Corp
Priority to JP2009070562A priority Critical patent/JP2010220525A/en
Publication of JP2010220525A publication Critical patent/JP2010220525A/en
Withdrawn legal-status Critical Current

Links

Images

Classifications

    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M47/00—Means for after-treatment of the produced biomass or of the fermentation or metabolic products, e.g. storage of biomass
    • C12M47/02—Separating microorganisms from the culture medium; Concentration of biomass
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M33/00—Means for introduction, transport, positioning, extraction, harvesting, peeling or sampling of biological material in or from the apparatus
    • C12M33/14—Means for introduction, transport, positioning, extraction, harvesting, peeling or sampling of biological material in or from the apparatus with filters, sieves or membranes

Landscapes

  • Life Sciences & Earth Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Wood Science & Technology (AREA)
  • Zoology (AREA)
  • Organic Chemistry (AREA)
  • Chemical & Material Sciences (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Biotechnology (AREA)
  • Sustainable Development (AREA)
  • Microbiology (AREA)
  • Biochemistry (AREA)
  • General Engineering & Computer Science (AREA)
  • General Health & Medical Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Biomedical Technology (AREA)
  • Molecular Biology (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)

Abstract

<P>PROBLEM TO BE SOLVED: To enable a stem cell to be separated in high yield. <P>SOLUTION: The apparatus 1 for separating the stem cell includes a container 4, a first filter 5 for dividing inside spaces A1 and A2 of the container 4, and having many through holes having bore diameters allowing the stem cell to pass, a liquid-discharging opening 3 opened to one space A2 divided by the first filter 5, and capable of being opened and closed, and a second filter 6 arranged at a position covering the liquid-discharging opening 3, and having many through holes having bore diameters prohibiting the stem cell to pass and allowing the liquid to pass. An inducing medium 11 containing an inducer for inducing angioplasty is filled in the space A2 held by the first and second filters 5 and 6 in the apparatus 1. <P>COPYRIGHT: (C)2011,JPO&INPIT

Description

本発明は、幹細胞分離装置および幹細胞分離方法に関するものである。   The present invention relates to a stem cell separation device and a stem cell separation method.

従来、骨髄などの幹細胞を有する組織から幹細胞を含む細胞群または幹細胞を抽出し、様々な疾患に対して細胞を投与、または患部に注入する細胞治療が行われている。このような幹細胞を抽出するための技術として、ＣＤ３４抗体で分離する技術が知られている（例えば、特許文献１、非特許文献１参照。）。   Conventionally, cell therapy including extracting stem cells or stem cells containing stem cells from a tissue having stem cells such as bone marrow and administering or injecting cells into various affected areas has been performed. As a technique for extracting such stem cells, a technique of separating with a CD34 antibody is known (see, for example, Patent Document 1 and Non-Patent Document 1).

米国特許第５５３６４７５号明細書US Pat. No. 5,536,475

臨床病理レビュー 特集第１２２号 「最新 造血細胞移植とその実際手技」、ｐ１４６−１５６Clinical Pathology Review Special Issue No. 122 “Latest Hematopoietic Cell Transplantation and its Actual Technique”, p146-156

しかしながら、ＣＤ３４抗体による分離は、収率が低いという問題がある。
本発明は、上述した事情に鑑みてなされたものであって、高い収率で幹細胞を分離することができる幹細胞分離装置および幹細胞分離方法を提供することを目的としている。 However, the separation using the CD34 antibody has a problem that the yield is low.
The present invention has been made in view of the above-described circumstances, and an object thereof is to provide a stem cell separation device and a stem cell separation method that can separate stem cells with high yield.

上記目的を達成するために、本発明は以下の手段を提供する。
本発明は、容器と、該容器の内部空間を区画し、幹細胞を通過可能な口径の多数の透孔を有する第１のフィルタと、該第１のフィルタによって区画された一方の空間に開口する開閉可能な液体排出口と、該液体排出口を覆う位置に配置され、幹細胞の通過を禁止し液体の通過を許容する口径の多数の透孔を有する第２のフィルタとを備え、前記第１および第２のフィルタに挟まれた空間に、血管形成を誘導する誘導因子を含む誘導培地が充填されている幹細胞分離装置を提供する。 In order to achieve the above object, the present invention provides the following means.
The present invention opens to a container, a first filter that partitions an internal space of the container, and has a large number of through-holes that can pass through stem cells, and one space partitioned by the first filter. A liquid discharge port that can be opened and closed; and a second filter that is disposed at a position covering the liquid discharge port and has a large number of through holes having a diameter that prohibits passage of stem cells and allows passage of liquid. And a stem cell separation device in which a space sandwiched between second filters is filled with an induction medium containing an inducer that induces angiogenesis.

本発明によれば、幹細胞を含む細胞懸濁液が容器内の第１のフィルタによって区画された空間に収容されると、細胞懸濁液が第１のフィルタを挟んで誘導培地に隣接する。誘導培地内に含まれている血管形成を誘導する誘導因子が、第１のフィルタを越えて細胞懸濁液に作用し、細胞懸濁液内に含まれている幹細胞が誘導培地の収容されている空間の方向に誘導される。   According to the present invention, when a cell suspension containing stem cells is accommodated in the space defined by the first filter in the container, the cell suspension is adjacent to the induction medium with the first filter interposed therebetween. An inducer that induces angiogenesis contained in the induction medium acts on the cell suspension beyond the first filter, and the stem cells contained in the cell suspension are contained in the induction medium. It is guided in the direction of the space.

第１のフィルタは幹細胞を通過させることができる口径の多数の透孔を有しているので、誘導された幹細胞は第１のフィルタの透孔を通過して誘導培地の空間内に入る。この状態で、残った細胞懸濁液を排出し、また、液体排出口を開放して第２のフィルタを介して誘導培地を排出することにより、誘導培地内に誘導されていた幹細胞を中央空間に残した状態で回収することができる。これにより、ＣＤ３４抗体によることなく、高い収率で幹細胞を分離することができる。
上記発明においては、前記誘導因子が、ＳＤＦ−１、ＭＣＰ−１、ＶＥＧＦまたはＨＧＦの少なくとも１つであることが好ましい。 Since the first filter has a large number of pores having a diameter that allows the stem cells to pass therethrough, the induced stem cells pass through the pores of the first filter and enter the space of the induction medium. In this state, the remaining cell suspension is discharged, and the stem cell that has been induced in the induction medium is removed from the central space by opening the liquid discharge port and discharging the induction medium through the second filter. It can be recovered in the state left in Thereby, a stem cell can be isolate | separated with a high yield, without depending on CD34 antibody.
In the said invention, it is preferable that the said induction | guidance | derivation factor is at least 1 of SDF-1, MCP-1, VEGF, or HGF.

また、本発明は、容器内に設けられた幹細胞を通過可能な口径の多数の透孔を有する第１のフィルタと、幹細胞の通過を禁止し液体の通過を許容する口径の多数の透孔を有する第２のフィルタとの間に挟まれる培地空間に、血管形成を誘導する誘導因子を含む誘導培地を充填しておき、前記培地空間に前記第１のフィルタを挟んで隣接する空間に、幹細胞を含む細胞懸濁液を収容して培養し、その後に、前記培地空間に開口し前記第２のフィルタによって覆われる液体排出口から容器の外部に誘導培地を排出する幹細胞分離方法を提供する。
上記発明においては、前記誘導因子が、ＳＤＦ−１、ＭＣＰ−１、ＶＥＧＦまたはＨＧＦの少なくとも１つであってもよい。 The present invention also includes a first filter having a large number of through holes provided in a container and capable of passing stem cells, and a plurality of through holes having a diameter that prohibits passage of stem cells and allows passage of liquid. An inducing medium containing an inducing factor for inducing angiogenesis is filled in the medium space sandwiched between the second filter and the stem cells in the space adjacent to the medium space with the first filter interposed therebetween. A stem cell separation method is provided in which a cell suspension containing is cultured and then the induction medium is discharged to the outside of the container from a liquid outlet that opens into the medium space and is covered by the second filter.
In the above invention, the inducer may be at least one of SDF-1, MCP-1, VEGF, or HGF.

本発明によれば、高い収率で幹細胞を分離することができるという効果を奏する。   According to the present invention, there is an effect that stem cells can be separated with high yield.

本発明の一実施形態に係る細胞分離装置を示す模式的な縦断面図である。It is a typical longitudinal section showing a cell separation device concerning one embodiment of the present invention. 図１の細胞分離装置による細胞分離方法を説明するフローチャートである。It is a flowchart explaining the cell separation method by the cell separation apparatus of FIG. 図１の細胞分離装置の上部空間に細胞懸濁液を導入した状態を示す模式的な縦断面図である。It is a typical longitudinal cross-sectional view which shows the state which introduced the cell suspension into the upper space of the cell separation apparatus of FIG. 図３の細胞分離装置により培養した後の状態を示す模式的な縦断面図である。It is a typical longitudinal cross-sectional view which shows the state after culture | cultivating with the cell separation apparatus of FIG. 図４の状態から上部空間の細胞懸濁液を排出した状態を示す模式的な縦断面図である。It is a typical longitudinal cross-sectional view which shows the state which discharged | emitted the cell suspension of upper space from the state of FIG. 図５の状態から誘導培地を排出した状態を示す模式的な縦断面図である。It is a typical longitudinal cross-sectional view which shows the state which discharged | emitted the induction culture medium from the state of FIG. 図６の状態から中央空間に洗浄液を供給した状態を示す模式的な縦断面図である。It is a typical longitudinal cross-sectional view which shows the state which supplied the washing | cleaning liquid to the central space from the state of FIG. 図７の状態から洗浄液を排出した状態を示す模式的な縦断面図である。It is a typical longitudinal cross-sectional view which shows the state which discharged | emitted the cleaning liquid from the state of FIG. 図８の状態から中央空間に乳酸リンゲル液を供給した状態を示す模式的な縦断面図である。It is a typical longitudinal cross-sectional view which shows the state which supplied the lactated Ringer's solution to the center space from the state of FIG. 図９の状態から中央空間から幹細胞を含む乳酸リンゲル液を吸引して回収する状態を説明する模式的な縦断面図である。It is a typical longitudinal cross-sectional view explaining the state which attracts | sucks and collect | recovers the lactate Ringer's solution containing a stem cell from the center space from the state of FIG. 図１の細胞分離装置において、誘導培地内のＳＤＦ−１の濃度と回収される細胞数との関係を示すグラフである。In the cell separation device of FIG. 1, it is a graph which shows the relationship between the density | concentration of SDF-1 in an induction culture medium, and the cell number collect | recovered.

本発明の一実施形態に係る細胞分離装置１および細胞分離方法について、図面を参照して以下に説明する。
本実施形態に係る細胞分離装置１は、図１に示されるように、上端に細胞懸濁液の導入口２を有し、下端に液体の第１の排出口３を有する筒状の容器４と、容器４の内部に上下方向に間隔をあけて配置され、容器４内部を上部空間Ａ１、中央空間Ａ２および下部空間Ａ３に区画する第１，第２のフィルタ５，６とを備えている。 A cell separation device 1 and a cell separation method according to an embodiment of the present invention will be described below with reference to the drawings.
As shown in FIG. 1, the cell separation apparatus 1 according to the present embodiment has a cylindrical container 4 having a cell suspension inlet 2 at the upper end and a liquid first outlet 3 at the lower end. And first and second filters 5 and 6 which are arranged inside the container 4 with a space in the vertical direction and partition the inside of the container 4 into an upper space A1, a central space A2 and a lower space A3. .

第１のフィルタ５は、幹細胞の通過を許容し、それよりも大きな細胞等の通過を禁止する程度の口径約８μｍの複数の透孔（図示略）を有している。
第２のフィルタ６は、幹細胞の通過を禁止し、液体の通過を許容する程度の口径約４μｍの複数の透孔（図示略）を有している。 The first filter 5 has a plurality of through-holes (not shown) having a diameter of about 8 μm that allow passage of stem cells and prohibit passage of larger cells and the like.
The second filter 6 has a plurality of through-holes (not shown) having a diameter of about 4 μm that prohibit passage of stem cells and allow passage of liquid.

上部空間Ａ１には、さらに、外気の流通を可能にしつつ、塵埃やウィルス等の容器４内への浸入を禁止するフィルタ７と、該上部空間Ａ１内の液体を排出するための開閉可能な第２の排出口８が設けられている。
また、中央空間Ａ２には、該中央空間Ａ２内に液体を供給するための液体供給口９と、最終的に中央空間Ａ２内に残留した幹細胞Ｂを回収するための回収口１０とが設けられている。 The upper space A1 further includes a filter 7 that allows the outside air to circulate while prohibiting the entry of dust and viruses into the container 4 and an openable and closable first for discharging the liquid in the upper space A1. Two outlets 8 are provided.
Further, the central space A2 is provided with a liquid supply port 9 for supplying a liquid into the central space A2, and a recovery port 10 for recovering the stem cells B finally remaining in the central space A2. ing.

容器４内の中央空間Ａ２および下部空間Ａ３には、誘導培地１１が収容されている。誘導培地１１は、例えば、ＤＭＥＭ／Ｆ１２培地に１０％血清および１００ｎｇ／ｍＬの血管形成を誘導する因子、例えば、ＳＤＦ−１、ＭＣＰ−１、ＶＥＧＦまたはＨＧＦの少なくとも１つを含んだものである。誘導培地１１は、第１のフィルタ５に接触する位置まで中央空間Ａ２および下部空間Ａ３内に満たされている。   In the medium space A2 and the lower space A3 in the container 4, the induction medium 11 is accommodated. The induction medium 11 is, for example, a medium containing 10% serum and 100 ng / mL angiogenesis, for example, SDF-1, MCP-1, VEGF, or HGF in DMEM / F12 medium. . The induction medium 11 is filled in the central space A <b> 2 and the lower space A <b> 3 up to a position where it contacts the first filter 5.

このように構成された本実施形態に係る細胞分離装置１を用いた細胞分離方法について、以下に説明する。
本実施形態に係る細胞分離装置１を用いて細胞懸濁液Ｃから幹細胞Ｂを分離するには、容器４の全ての排出口３，８、回収口１０および供給口９を閉じた状態で、図１および図２に示されるように、中央空間Ａ２および下部空間Ａ３内に誘導培地１１を貯留する（ステップＳ１）。 A cell separation method using the cell separation device 1 according to the present embodiment configured as described above will be described below.
In order to separate the stem cells B from the cell suspension C using the cell separation device 1 according to the present embodiment, with all the discharge ports 3, 8, the collection ports 10 and the supply ports 9 closed, As shown in FIGS. 1 and 2, the induction medium 11 is stored in the central space A2 and the lower space A3 (step S1).

この状態で、容器４の上部の導入口２を開放し、図３に示されるように、導入口２から細胞懸濁液Ｃを容器４の上部空間Ａ１内に導入する（ステップＳ２）。
細胞懸濁液Ｃは、例えば、骨髄液を乳酸リンゲル液等によって希釈したもので、幹細胞Ｂの他、種々の細胞を含んでいる。 In this state, the inlet 2 at the top of the container 4 is opened, and the cell suspension C is introduced from the inlet 2 into the upper space A1 of the container 4 as shown in FIG. 3 (step S2).
The cell suspension C is, for example, a bone marrow fluid diluted with lactated Ringer's solution or the like, and includes various cells in addition to the stem cell B.

上部空間Ａ１内に導入された細胞懸濁液Ｃは、第１のフィルタ５を挟んで中央空間Ａ２内の誘導培地１１に隣接させられる。
この状態で、図４に示されるように、所定の培養条件、例えば、３７℃の環境下に容器４ごと配置して、静置することにより、細胞懸濁液Ｃ内の幹細胞Ｂを培養する（ステップＳ３）。 The cell suspension C introduced into the upper space A1 is made adjacent to the induction medium 11 in the central space A2 with the first filter 5 interposed therebetween.
In this state, as shown in FIG. 4, the stem cell B in the cell suspension C is cultured by placing the container 4 together in a predetermined culture condition, for example, an environment of 37 ° C. and allowing it to stand. (Step S3).

誘導培地１１には、血管形成を誘導する因子が含まれているので、この因子の作用によって、細胞懸濁液Ｃ内の幹細胞Ｂが細胞懸濁液Ｃ内を遊走して中央空間Ａ２の方向に引き寄せられる。他の細胞にはこの因子はほぼ作用しない。   Since the induction medium 11 contains a factor that induces angiogenesis, the stem cell B in the cell suspension C migrates in the cell suspension C by the action of this factor, and the direction of the central space A2 Be drawn to. This factor has little effect on other cells.

そして、中央空間Ａ２の方向に引き寄せられた幹細胞Ｂは、時間の経過によって、第１のフィルタ５の透孔を通過して中央空間Ａ２の誘導培地１１内に入る。
所定時間、例えば、２２時間経過した後に、図５に示されるように、上部空間Ａ１に設けられた第２の排出口８を開放して、細胞懸濁液Ｃを排出する（ステップＳ４）。次いで、図６に示されるように、容器４の下部の第１の排出口３を開放して、中央空間Ａ２および下部空間Ａ３に貯留されている誘導培地１１を排出する（ステップＳ５）。 Then, the stem cells B attracted in the direction of the central space A2 pass through the through holes of the first filter 5 and enter the induction medium 11 in the central space A2 over time.
After a predetermined time, for example, 22 hours have elapsed, as shown in FIG. 5, the second discharge port 8 provided in the upper space A1 is opened to discharge the cell suspension C (step S4). Next, as shown in FIG. 6, the first discharge port 3 at the bottom of the container 4 is opened, and the induction medium 11 stored in the central space A2 and the lower space A3 is discharged (step S5).

第２のフィルタ６は、幹細胞Ｂの通過を禁止する口径の透孔を有しているので、中央空間Ａ２内に誘導されてきていた幹細胞Ｂは、第２のフィルタ６によって捕捉され、誘導培地１１のみが第１の排出口３から排出される。
そして、誘導培地１１が排出され終わったら、第１の排出口３を閉じて、図７に示されるように、中央空間Ａ２に設けられた液体供給口９から中央空間Ａ２内に洗浄液Ｄを供給し（ステップＳ６）、第２のフィルタ６に捕捉されていた幹細胞Ｂを再懸濁する。そして、図８に示されるように、第１の排出口３を開放して、洗浄液Ｄを排出する（ステップＳ７）。 Since the second filter 6 has a through-hole having a diameter that prohibits passage of the stem cell B, the stem cell B that has been induced in the central space A2 is captured by the second filter 6 and is induced medium. Only 11 is discharged from the first outlet 3.
When the induction medium 11 is completely discharged, the first discharge port 3 is closed, and the cleaning liquid D is supplied into the central space A2 from the liquid supply port 9 provided in the central space A2, as shown in FIG. Then, the stem cell B captured by the second filter 6 is resuspended (step S6). And as FIG. 8 shows, the 1st discharge port 3 is open | released and the washing | cleaning liquid D is discharged | emitted (step S7).

必要により、ステップＳ６およびステップＳ７を繰り返すことにより、第２のフィルタ６に捕捉された幹細胞Ｂを洗浄する（ステップＳ８）。この後に、図９に示されるように、液体供給口９から乳酸リンゲル液のような回収用の液体Ｅを中央空間Ａ２内に供給して幹細胞Ｂを再懸濁し（ステップＳ９）、図１０に示されるように、回収口１０に接続したシリンジ１２等によって幹細胞Ｂを含む液体Ｅを回収する（ステップＳ１０）。   If necessary, the stem cells B captured by the second filter 6 are washed by repeating Step S6 and Step S7 (Step S8). Thereafter, as shown in FIG. 9, a recovery liquid E such as lactated Ringer's solution is supplied from the liquid supply port 9 into the central space A2 to resuspend the stem cells B (step S9), as shown in FIG. As described above, the liquid E containing the stem cells B is collected by the syringe 12 or the like connected to the collection port 10 (step S10).

このように、本実施形態に係る細胞分離装置１および細胞分離方法によれば、上部空間Ａ１に幹細胞Ｂを含む細胞懸濁液Ｃを貯留して、所定の時間にわたって培養するだけで、細胞懸濁液Ｃ内の幹細胞Ｂを中央空間Ａ２に遊走させて分離することができる。したがって、ＣＤ３４抗体を用いることなく、高い収率で、幹細胞Ｂを回収することができるという利点がある。   As described above, according to the cell separation device 1 and the cell separation method according to the present embodiment, the cell suspension C containing the stem cells B is stored in the upper space A1 and cultured for a predetermined time. Stem cells B in the suspension C can migrate to the central space A2 and can be separated. Therefore, there is an advantage that the stem cells B can be recovered with a high yield without using the CD34 antibody.

なお、本実施形態においては、ＳＤＦ−１の濃度として、１００ｎｇ／ｍＬを例示したが、これに限定されるものではない。図１１に示されるように、ＳＤＦ−１の濃度と回収される細胞数との関係を示す。図１１によれば、ＳＤＦ−１を含まない場合と比較して、５０ｎｇ／ｍＬ以上のＳＤＦ−１が含有されていれば、いずれの場合も細胞回収率が増大していることがわかる。   In addition, in this embodiment, although 100 ng / mL was illustrated as a density | concentration of SDF-1, it is not limited to this. As shown in FIG. 11, the relationship between the concentration of SDF-1 and the number of recovered cells is shown. According to FIG. 11, it can be seen that the cell recovery rate is increased in any case as long as 50 ng / mL or more of SDF-1 is contained as compared with the case where SDF-1 is not included.

１ 幹細胞分離装置
３ 第１の排出口（液体排出口）
４ 容器
５ 第１のフィルタ
６ 第２のフィルタ
１１ 誘導培地
Ａ１ 上部空間（内部空間）
Ａ２ 中央空間（空間）
Ｂ 幹細胞 1 Stem cell separation device 3 First outlet (liquid outlet)
4 container 5 first filter 6 second filter 11 induction medium A1 upper space (internal space)
A2 Central space (space)
B stem cells

Claims (4)

容器と、
該容器の内部空間を区画し、幹細胞を通過可能な口径の多数の透孔を有する第１のフィルタと、
該第１のフィルタによって区画された一方の空間に開口する開閉可能な液体排出口と、
該液体排出口を覆う位置に配置され、幹細胞の通過を禁止し液体の通過を許容する口径の多数の透孔を有する第２のフィルタとを備え、
前記第１および第２のフィルタに挟まれた空間に、血管形成を誘導する誘導因子を含む誘導培地が充填されている幹細胞分離装置。 A container,
A first filter that partitions the internal space of the container and has a large number of through-holes that can pass through stem cells;
An openable and closable liquid outlet opening into one space partitioned by the first filter;
A second filter that is disposed at a position covering the liquid outlet and has a large number of through holes having a diameter that prohibits passage of stem cells and allows passage of liquid;
A stem cell separation device, wherein a space sandwiched between the first and second filters is filled with an induction medium containing an inducer that induces angiogenesis. 前記誘導因子が、ＳＤＦ−１、ＭＣＰ−１、ＶＥＧＦまたはＨＧＦの少なくとも１つである請求項１に記載の幹細胞分離装置。   The stem cell separation device according to claim 1, wherein the inducer is at least one of SDF-1, MCP-1, VEGF, or HGF. 容器内に設けられた幹細胞を通過可能な口径の多数の透孔を有する第１のフィルタと、幹細胞の通過を禁止し液体の通過を許容する口径の多数の透孔を有する第２のフィルタとの間に挟まれる培地空間に、血管形成を誘導する誘導因子を含む誘導培地を充填し、
前記培地空間に前記第１のフィルタを挟んで隣接する空間に、幹細胞を含む細胞懸濁液を収容して培養し、
その後に、前記培地空間に開口し前記第２のフィルタによって覆われる液体排出口から容器の外部に誘導培地を排出する幹細胞分離方法。 A first filter having a large number of through-holes having a diameter capable of passing stem cells provided in a container, and a second filter having a large number of through-holes having a diameter that prohibits passage of stem cells and allows passage of liquid; Fill the medium space sandwiched between the induction medium containing the inducer that induces angiogenesis,
Accommodating and culturing a cell suspension containing stem cells in a space adjacent to the medium space across the first filter,
Then, the stem cell separation method of discharging the induction medium to the outside of the container from a liquid discharge port that opens into the medium space and is covered by the second filter. 前記誘導因子が、ＳＤＦ−１、ＭＣＰ−１、ＶＥＧＦまたはＨＧＦの少なくとも１つである請求項３に記載の幹細胞分離方法。   The stem cell separation method according to claim 3, wherein the inducer is at least one of SDF-1, MCP-1, VEGF, or HGF.
JP2009070562A 2009-03-23 2009-03-23 Apparatus and method for separating stem cell Withdrawn JP2010220525A (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
JP2009070562A JP2010220525A (en) 2009-03-23 2009-03-23 Apparatus and method for separating stem cell

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
JP2009070562A JP2010220525A (en) 2009-03-23 2009-03-23 Apparatus and method for separating stem cell

Publications (1)

Publication Number Publication Date
JP2010220525A true JP2010220525A (en) 2010-10-07

Family

ID=43038421

Family Applications (1)

Application Number Title Priority Date Filing Date
JP2009070562A Withdrawn JP2010220525A (en) 2009-03-23 2009-03-23 Apparatus and method for separating stem cell

Country Status (1)

Country Link
JP (1) JP2010220525A (en)

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2017523770A (en) * 2015-06-10 2017-08-24 ヒューマン メッド アーゲー Device for separating regenerative and adult stem cells

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2017523770A (en) * 2015-06-10 2017-08-24 ヒューマン メッド アーゲー Device for separating regenerative and adult stem cells

Similar Documents

Publication Publication Date Title
US10537596B2 (en) Bone marrow adipose portion isolation device and methods
JP6364946B2 (en) Separation structure and separation method
JP2021097690A (en) Cell separation device and method for using the same
JP2012510272A5 (en)
US10697964B2 (en) Liquid biopsy detection of leukemia using closed-loop microfluidics
CN110325632B (en) Apparatus and method for separating mobile cells
RU2708206C2 (en) Particle treatment
US9459189B2 (en) Device for isolating a fraction in a biological sample
CN107109323A (en) Nucleic acid-extracting apparatus and its method of work
Shin et al. Integrative Magneto‐Microfluidic Separation of Immune Cells Facilitates Clinical Functional Assays
JP2010220525A (en) Apparatus and method for separating stem cell
CN106660043A (en) Device and method for cell nuclei preparation
US9713810B2 (en) Cell washing plunger using centrifugal force
CN107917833A (en) The method for recycling rare cell
US20120261348A1 (en) Apparatus for extracting biomaterial and method of extracting biomaterial using the apparatus
KR101313048B1 (en) Dividing device for cell fractions or blood elements
CN114632564A (en) Integrated micro-fluidic chip and in-vitro treatment method for primary circulating tumor cells
JP5486441B2 (en) Virus collection system
ES2936343T3 (en) Device for dividing objects and dividing procedure
WO2018187181A1 (en) Microfluidic system for evaluation of chemotherapeutic and immunotherapeutic drugs
JP2006246835A (en) Separation/recovery container for density gradient
KR101576120B1 (en) Filtering apparatus for liquid
CN108220233A (en) Cell separation set surface treatment method, related utensil, peripheral blood rare cell or circulating tumor cell rapidly and efficiently separation method
JP2008212022A (en) Device and method for separating stem cells originated from tissues
CN205603580U (en) Rare cell enrichment filter equipment

Legal Events

Date Code Title Description
A300 Withdrawal of application because of no request for examination

Free format text: JAPANESE INTERMEDIATE CODE: A300

Effective date: 20120605