WO2021114087A1 - 小体积细胞培养袋 - Google Patents

小体积细胞培养袋 Download PDF

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Publication number
WO2021114087A1
WO2021114087A1 PCT/CN2019/124301 CN2019124301W WO2021114087A1 WO 2021114087 A1 WO2021114087 A1 WO 2021114087A1 CN 2019124301 W CN2019124301 W CN 2019124301W WO 2021114087 A1 WO2021114087 A1 WO 2021114087A1
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cell culture
closed
bag
bag body
small
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PCT/CN2019/124301
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English (en)
French (fr)
Inventor
张豪
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苏州生动细胞生物科技有限公司
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Priority to PCT/CN2019/124301 priority Critical patent/WO2021114087A1/zh
Publication of WO2021114087A1 publication Critical patent/WO2021114087A1/zh

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    • 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
    • C12M1/00—Apparatus for enzymology or microbiology

Definitions

  • the present application relates to the field of cell culture technology, in particular to a small-volume cell culture bag.
  • the cell culture bag is a disposable device when culturing cells. According to actual needs, the size and volume of the cell culture bag are variable or selectable.
  • the common cell culture bag is composed of two upper and lower plastic layers of special materials ( That is, the upper bag piece and the lower bag piece) form a flat cell culture bag after hot pressing.
  • the minimum culture volume (culture medium volume) of a culture bag is equal to the bottom area of the bag.
  • the maximum working volume is proportional to the height of the bulge when the culture bag is inflated, the higher the bulge, the larger the maximum working volume.
  • the common cell bag is a 2D bag, and the bulging height is basically determined after the bottom area is determined.
  • the bulging height of the flat cell bag after inflating is too low, which makes the space above the liquid insufficient.
  • the liquid wave will easily block the air inlet and outlet, which affects the control of air pressure, and ultimately will directly affect the cultured cells; for common cell culture bags on the market The smallest is 1L bag), the above influence will be reduced, but for small-volume cell culture bags, such as small-volume cell culture bags with a culture volume of less than 300ml, the 2D flat structure of the bag body will severely limit the minimum and The range of the maximum working volume.
  • common 2D bags when the minimum culture volume is 30ml, the maximum culture volume is only 100ml.
  • the purpose of this application is to overcome the limitation of the minimum and maximum working volume when the existing cell culture bag is in a flat state due to the small volume of the cell culture bag, and to provide a small-volume cell culture bag including a closed bag body.
  • a joint is welded or thermally welded to the bending portion, and the joint includes a plate connected to the closed bag body for supporting the closed bag body.
  • a vertical or inclined flat portion is provided at the bending position, and the flat portion is formed after the bag body is closed by heat fusion and cooled.
  • the entrance and exit of the bending part are provided on the joint or the flat part.
  • the outer side to the inner side of the closed bag body includes a PA layer, an EVOH layer, a PE layer and a ULDPE layer in sequence, and an adhesive resin layer is arranged between each layer.
  • entrance and exit are round holes.
  • the bending angle of the bending part is less than 180 degrees.
  • the flat closed bag can be supported by the support of the bend, so that the closed bag has a non-flat structure, which provides longitudinal space for cell culture, facilitates the control of air pressure and temperature, and is beneficial to cells Cultivate and observe.
  • Figure 1 is a schematic diagram of the structure of a small-volume cell culture bag in this application.
  • Fig. 2 is a partial three-dimensional structure diagram taken along the A-A section of Fig. 1.
  • FIG. 3 is a schematic diagram of the structure with a flat part provided at the bend in FIG. 2.
  • Fig. 4 is a schematic diagram of the structure of Fig. 2 with a joint at the bend.
  • the small-volume cell culture bag shown in Figures 1 to 4 includes a closed bag body 1.
  • the closed bag body 1 is a closed bag cavity formed by bending a single piece of bag and hot pressing.
  • the single piece of bag After bending once with its midline, the other three sides except the bend 2 are hot-pressed to form a closed pocket; the traditional cell culture bag usually consists of the upper and lower pockets through a single hot pressing Four sides are formed.
  • This kind of culture bag has a flat bag structure, which causes the cells to be easily pressed by the flat culture bag, causing cell loss when the cells flow.
  • the closed bag body 1 of this embodiment has one side as a bend 2 (Also the bent side), and the bending angle ⁇ of the bending part 2 is less than 180 degrees.
  • the bending angle ⁇ shown in Figure 2 in this embodiment is 150 degrees, so the upper and lower layers of the closed bag body 1 can be prevented from being stuck together , So that at least a part of the closed bag body 1 is propped up to form a three-dimensional structure instead of a flat bag structure, which is more conducive to cell culture and observation; of course, the above-mentioned molding method is not limited to thermocompression molding, and can also be other conventional Forming method; the closed bag body 1 is provided with a plurality of inlets and outlets 3 for liquid or gas in and out, and at least one inlet and outlet 3 is provided at the bend 2 of the closed bag body 1. In this embodiment, there are two entrances and exits 3 at the bend 2.
  • the support can be made by heat melting.
  • the tube body 5 and the disc body 4 can be an integral structure or a separate assembly structure; the tube body 5 can also be connected to other inlets and outlets 3 in a sealed manner.
  • the entrance and exit 3 of the bending part 2 are directly provided on the joint or flat part 6.
  • the single bag piece of this embodiment should have a certain degree of toughness, and the bending part 2 will not break or collapse during bending.
  • the outer side to the inner side of the closed bag body 1 are sequentially arranged with PA (polyamide-polyamide-) with a thickness of 24um.
  • Nylon preferably an EVOH (ethylene/vinyl alcohol copolymer) layer with a thickness of 8um, a PE (polyethylene) layer with a thickness of 58um and a ULDPE (ultra-low density polyethylene) layer with a thickness of 260um, and a PA layer , 5-15um thick adhesive resin (Tie) layer is arranged between the EVOH layer, the PE layer and the ULDPE layer. Due to the use of multi-layer biological sealed packaging, no additional sterilization or cleaning is required before use.
  • the entrance and exit 3 of this embodiment are preferably round holes.
  • the volume of the closed bag body 1 may be 10-2000 ml, preferably 20-200 ml.
  • the small-volume cell culture bag in this embodiment refers to a cell culture bag with a minimum culture volume of 30 ml and a maximum culture volume of 300 ml.

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  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Health & Medical Sciences (AREA)
  • Wood Science & Technology (AREA)
  • Biotechnology (AREA)
  • Organic Chemistry (AREA)
  • Zoology (AREA)
  • Microbiology (AREA)
  • Sustainable Development (AREA)
  • Biomedical Technology (AREA)
  • Biochemistry (AREA)
  • General Engineering & Computer Science (AREA)
  • General Health & Medical Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Medicinal Chemistry (AREA)
  • Apparatus Associated With Microorganisms And Enzymes (AREA)

Abstract

小体积细胞培养袋，包括封闭袋体，所述封闭袋体是由单块袋片折弯后热压形成的一个封闭袋腔；所述封闭袋体上设有容液体或气体进出的多个出入口，至少一个所述出入口设于所述封闭袋体的折弯处。通过折弯处的支撑作用，可将平整的封闭袋体支撑开，使封闭袋体为非平整结构，为细胞培养提供纵向空间，从而提高更广的最小和最大工作体积的范围。

Description

小体积细胞培养袋 技术领域
本本申请涉及细胞培养技术领域，尤其涉及一种小体积细胞培养袋。
背景技术
细胞培养袋在培养细胞时是一次性的装置，根据实际需求，细胞培养袋的大小、体积都是可变的或者可以选择的，常见的细胞培养袋是由上下两片特殊材质的塑料层(即上层袋片和下层袋片)经过热压形成的一个平整的细胞培养袋，在对细胞培养袋进行充气或进液时，一个培养袋的最小培养体积(培养液体积)跟袋底面积成正比，底面积越小，最小工作体积就越小；最大工作体积和培养袋充气时鼓起的高度成正比，鼓起越高，最大工作体积越大。但是常见的细胞袋是2D袋，底面积确定后鼓起高度基本也确定。平整细胞袋充气后鼓起的高度太低，使液体上方的空间不够，摇晃时液浪容易堵塞进出气口，影响气压的控制，最终都将直接影响培养的细胞；对于市场上常见细胞培养袋(最小的也是1L袋)而言，上述影响会有所减小，但对于小体积细胞培养袋而言，比如培养容积小于300ml的小体积细胞培养袋，袋体的2D平整结构将严重限制最小和最大工作体积的范围。常见2D袋，当最小培养体积是30ml时，最大培养体积只有100ml。
发明内容
本申请的目的在于克服现有细胞培养袋体积较小时由于袋体处于平整状态会限制最小和最大工作体积的范围，提供一种小体积细胞培养袋，包括封闭袋体，所述封闭袋体是由单块袋片折弯后热 压形成的一个封闭袋腔；所述封闭袋体上设有容液体或气体进出的多个出入口，至少一个所述出入口设于所述封闭袋体的折弯处。
进一步，所述折弯处焊接或热熔连接有接头，所述接头包括与封闭袋体连接的用于撑起封闭袋体的盘体。
进一步，所述折弯处设有竖直状或倾斜状的扁平部，所述扁平部通过热熔封闭袋体并冷却后形成。
进一步，所述折弯处的出入口设于所述接头或扁平部上。
进一步，所述封闭袋体的外侧至内侧依次包括PA层、EVOH层、PE层和ULDPE层，且各层之间设有粘合树脂层。
进一步，所述出入口为圆孔。
进一步，所述折弯处的弯曲角度小于180度。
本申请的有益效果是：通过折弯处的支撑作用，可将平整的封闭袋体支撑开，使封闭袋体为非平整结构，为细胞培养提供纵向空间，便于气压和温度的控制，利于细胞培养和观察。
附图说明
图1为本申请小体积细胞培养袋的结构示意图。
图2为图1的A-A剖视局部立体结构示意图。
图3为图2中折弯处设有扁平部的结构示意图。
图4为图2中折弯处设有接头的结构示意图。
图中，1.封闭袋体；2.折弯处；3.出入口；4.盘体；5.管体；6.扁平部。
具体实施方式
以下结合附图和具体实施例对本申请作进一步的详细描述。
如图1～4所示的小体积细胞培养袋，包括封闭袋体1，封闭袋体1是由单块袋片折弯后热压形成的一个封闭袋腔，本实施例中单块袋片以其中线折弯一次后，对除折弯处2外的另外三边进行热压 成型，最终形成一个封闭袋腔；传统的细胞培养袋通常是由上层袋片和下层袋片通过一次热压四条边成型，这种培养袋为平整的袋结构，导致细胞容易被平整的培养袋压住，其细胞进行流动时造成细胞损失，而本实施例的封闭袋体1有一边为折弯处2(也为折弯边)，且折弯处2的弯曲角度α小于180度，本实施例中图2所示的弯曲角度α为150度，因此可以避免封闭袋体1的上层和下层贴合，使封闭袋体1至少一部分被撑起从而形成立体结构而非平整的袋结构，更利于细胞培养和观察；当然上述的这种成型方式也并非仅限定热压成型，也可以是其他常规的成型方式；封闭袋体1上设有容液体或气体进出的多个出入口3，至少一个出入口3设于封闭袋体1的折弯处2。本实施例中折弯处2设有两个出入口3。
为了进一步防止封闭袋体1的上层和下层贴合在一起，需要在折弯处2设置支撑件以撑开封闭袋体1的上层和下层，如图3所示，支撑件可以是通过热熔封闭袋体1的折弯处2并冷却后形成的竖直状或倾斜状的扁平部6；支撑件也可以是焊接或热熔连接在折弯处2的接头，如图4所示，接头包括与封闭袋体1连接的用于撑起封闭袋体1的盘体4；其中，盘体4也为竖直状或倾斜状，接头还可包括与盘体4连接的管体5，管体5用于传输气体或液体，管体5与盘体4可以是一体结构，也可以为分体组装结构；管体5也可与其他出入口3密封连接。
为了利于细胞培养，折弯处2的出入口3直接设于接头或扁平部6上。
本实施例的单块袋片应该具有一定的韧性，在折弯时折弯处2不会断裂或塌下，封闭袋体1的外侧至内侧依次设置的优选厚度为24um的PA(聚酰胺—尼龙)层，优选厚度为8um的EVOH(乙烯/乙烯醇共聚物)层，优选厚度为58um的PE(聚乙烯)层和优 选厚度为260um的ULDPE(超低密度聚乙烯)层，且PA层、EVOH层、PE层和ULDPE层之间设有5～15um厚的粘合树脂(Tie)层。由于使用多层生物级密封包装，使用前不需要额外进行灭菌或清洁。
本实施例的出入口3优选为为圆孔。封闭袋体1的容积可以为10～2000ml，优选为20～200ml。本实施例所述的小体积细胞培养袋即指最小培养体积为30ml同时最大培养体积为300ml的细胞培养袋。

Claims (7)

  1. 一种小体积细胞培养袋，其特征在于：包括封闭袋体(1)，所述封闭袋体(1)是由单块袋片折弯后热压形成的一个封闭袋腔；所述封闭袋体(1)上设有容液体或气体进出的多个出入口(3)，至少一个所述出入口(3)设于所述封闭袋体(1)的折弯处(2)。
  2. 根据权利要求1所述的小体积细胞培养袋，其特征在于：所述折弯处(2)焊接或热熔连接有接头，所述接头包括与封闭袋体(1)连接的用于撑起封闭袋体(1)的盘体(4)。
  3. 根据权利要求1所述的小体积细胞培养袋，其特征在于：所述折弯处(2)设有竖直状或倾斜状的扁平部(6)，所述扁平部(6)通过热熔封闭袋体(1)并冷却后形成。
  4. 根据权利要求2或3所述的小体积细胞培养袋，其特征在于：所述折弯处(2)的出入口(3)设于所述接头或扁平部(6)上。
  5. 根据权利要求1所述的小体积细胞培养袋，其特征在于：所述封闭袋体(1)的外侧至内侧依次包括PA层、EVOH层、PE层和ULDPE层，且各层之间设有粘合树脂层。
  6. 根据权利要求1所述的小体积细胞培养袋，其特征在于：所述出入口(3)为圆孔。
  7. 根据权利要求1所述的小体积细胞培养袋，其特征在于：所述折弯处(2)的弯曲角度小于180度。
PCT/CN2019/124301 2019-12-10 2019-12-10 小体积细胞培养袋 WO2021114087A1 (zh)

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* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20050186669A1 (en) * 2004-02-20 2005-08-25 Cesco Bioengineering Co., Ltd. Apparatus and method for preparing and culturing cells
US20080131960A1 (en) * 2006-11-15 2008-06-05 Millipore Corporation Self standing bioreactor construction
US20090107040A1 (en) * 2006-04-21 2009-04-30 Frans Vandenhove Recipient for containing moist substrate
FR2931484A1 (fr) * 2008-05-26 2009-11-27 Assist Publ Hopitaux De Paris Bioreacteur a double compartiment.
WO2011079180A1 (en) * 2009-12-23 2011-06-30 Ge Healthcare Bio-Sciences Corp. Improved bioreactors
CN102373150A (zh) * 2010-08-24 2012-03-14 新奥科技发展有限公司 光生物反应器和光生物培养系统
EP2495301A1 (en) * 2011-03-03 2012-09-05 Meissner Filtration Products, Inc. Biocontainer
WO2018182993A1 (en) * 2017-03-28 2018-10-04 Corning Incorporated Aseptic bioprocess package
WO2019075775A1 (zh) * 2017-10-19 2019-04-25 江阴升辉包装材料有限公司 一种适用于生产线自动快速包装的热收缩膜
WO2019086324A1 (en) * 2017-10-31 2019-05-09 Ge Healthcare Bio-Sciences Corp. A flexible bag
WO2019214465A1 (zh) * 2018-05-09 2019-11-14 武汉赛科成科技有限公司 一种填充片状载体的流化床细胞培养一次性膜装置

Patent Citations (11)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US20050186669A1 (en) * 2004-02-20 2005-08-25 Cesco Bioengineering Co., Ltd. Apparatus and method for preparing and culturing cells
US20090107040A1 (en) * 2006-04-21 2009-04-30 Frans Vandenhove Recipient for containing moist substrate
US20080131960A1 (en) * 2006-11-15 2008-06-05 Millipore Corporation Self standing bioreactor construction
FR2931484A1 (fr) * 2008-05-26 2009-11-27 Assist Publ Hopitaux De Paris Bioreacteur a double compartiment.
WO2011079180A1 (en) * 2009-12-23 2011-06-30 Ge Healthcare Bio-Sciences Corp. Improved bioreactors
CN102373150A (zh) * 2010-08-24 2012-03-14 新奥科技发展有限公司 光生物反应器和光生物培养系统
EP2495301A1 (en) * 2011-03-03 2012-09-05 Meissner Filtration Products, Inc. Biocontainer
WO2018182993A1 (en) * 2017-03-28 2018-10-04 Corning Incorporated Aseptic bioprocess package
WO2019075775A1 (zh) * 2017-10-19 2019-04-25 江阴升辉包装材料有限公司 一种适用于生产线自动快速包装的热收缩膜
WO2019086324A1 (en) * 2017-10-31 2019-05-09 Ge Healthcare Bio-Sciences Corp. A flexible bag
WO2019214465A1 (zh) * 2018-05-09 2019-11-14 武汉赛科成科技有限公司 一种填充片状载体的流化床细胞培养一次性膜装置

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