WO2018086537A1 - 细胞培养箱及其工作方法 - Google Patents

细胞培养箱及其工作方法 Download PDF

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WO2018086537A1
WO2018086537A1 PCT/CN2017/109953 CN2017109953W WO2018086537A1 WO 2018086537 A1 WO2018086537 A1 WO 2018086537A1 CN 2017109953 W CN2017109953 W CN 2017109953W WO 2018086537 A1 WO2018086537 A1 WO 2018086537A1
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cell culture
door
culture incubator
disposed
box
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PCT/CN2017/109953
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English (en)
French (fr)
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闻路红
洪欢欢
史振志
施露露
甘剑勤
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宁波华仪宁创智能科技有限公司
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Publication of WO2018086537A1 publication Critical patent/WO2018086537A1/zh
Priority to US16/257,341 priority Critical patent/US20190153379A1/en

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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
    • C12M41/00—Means for regulation, monitoring, measurement or control, e.g. flow regulation
    • C12M41/12—Means for regulation, monitoring, measurement or control, e.g. flow regulation of temperature
    • C12M41/14—Incubators; Climatic chambers
    • 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
    • C12M23/00—Constructional details, e.g. recesses, hinges
    • C12M23/34—Internal compartments or partitions
    • 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
    • C12M41/00—Means for regulation, monitoring, measurement or control, e.g. flow regulation
    • C12M41/12—Means for regulation, monitoring, measurement or control, e.g. flow regulation of temperature
    • 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
    • C12M41/00—Means for regulation, monitoring, measurement or control, e.g. flow regulation
    • C12M41/12—Means for regulation, monitoring, measurement or control, e.g. flow regulation of temperature
    • C12M41/18—Heat exchange systems, e.g. heat jackets or outer envelopes
    • C12M41/22—Heat exchange systems, e.g. heat jackets or outer envelopes in contact with the bioreactor walls

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  • the invention relates to cell culture, in particular to a cell culture incubator and a method for its operation.
  • the cell culture workstation provides a culture environment of ultra-clean and confined space (ISO 5 or GMP grade), and the built-in incubator is a special device for cells to simulate physiological environment in vivo. It should have temperature control characteristics such as constant temperature, high temperature uniformity, rapid temperature recovery, and condensed water phenomenon caused by no temperature difference.
  • the current incubator heating methods are:
  • the stability of the heating method depends largely on the stability and response time of the heating wire, that is, the reasonable thickness and diameter of the heating wire are selected to slow the internal consumption of the heating wire element, and it needs to be timed. Adding water, washing, emptying and monitoring the tank is particularly difficult for incubator design.
  • Air-sleeve type the heating method has poor heat uniformity, and the air passage and the air outlet of the inner circulation type internal cavity wall have high design requirements and high cost, and the outer circulation type structure is relatively simple, and the thermal conductivity is not All.
  • the heating method has high temperature control precision, and the six-sided heating method can basically eliminate the condensed water problem of the glass door, but the temperature field distribution in the waiting box is uneven, and the consistency of the cell culture environment is destroyed, and When one or more of the inner wall of the incubator is faulty, it has a great influence on the temperature control stability of the chamber, especially for the stability and continuity of the cell culture environment, such as replacement of the heating piece and replacement of parts. Difficulty.
  • the present invention provides a cell culture incubator with precise temperature control and good temperature uniformity.
  • a cell culture incubator comprising:
  • the second box body is disposed inside the first box body, and a closed space is formed between the second box body and the first box body;
  • the first box door is disposed on a front side portion of the first box body, and a sealing member is disposed between the first box body and the first box door;
  • thermo measuring device disposed in the second box
  • Heating the device being disposed on an inner side of the first door and a side of the second case;
  • the heat insulating device is disposed at an outer side portion of the first case or an inner side portion of the first case.
  • Another object of the present invention is to provide a cell culture workstation with high temperature control precision and uniform heating, and the object of the invention is achieved by the following technical solutions:
  • a cell culture workstation includes a cabinet, a console, and a glove hole, the glove hole being disposed on a front side of the cabinet; the cell culture workstation further comprising:
  • a cell culture incubator using the above-mentioned cell culture incubator, a front side portion of the first tank body is fixed on a rear side portion of the cabinet, and a rear side portion of the cabinet has a position with the first tank door Through holes that match the size.
  • the working method comprises the following steps:
  • the present invention has the following beneficial effects:
  • the temperature control accuracy in the incubator is higher and more accurate, the temperature field distribution is uniform, and the culture consistency effect is good;
  • the gas jacket heating constant temperature and the wrapped direct heat method effectively solve the condensed water of the transparent box door and the gas pipeline, the temperature control efficiency is high, and the temperature recovery is fast;
  • FIG. 1 is a schematic structural view of a cell culture incubator according to Embodiment 1 of the present invention.
  • Figure 2 is a cross-sectional view along line A-A of the cell culture incubator according to Example 1 of the present invention
  • Figure 3 is a schematic block diagram of a cell culture workstation according to Embodiment 2 of the present invention.
  • the cell culture incubator includes:
  • a first case 62 such as made of a polished electrolytic stainless steel material
  • a second box body 65 the second box body is disposed inside the first box body, and a closed space is formed between the second box body and the first box body;
  • first box door 61 the first box door is disposed on a front side portion 63 of the first box body, and the first box body a seal between the first door and the first door;
  • the second box door is disposed on a front side portion of the first box body and is inside the first box door, and the second box door is made of a transparent material such as glass
  • the second door has at least two through holes, and transparent doors 642-643 that block the through holes;
  • a temperature measuring device 69 such as a temperature sensor, the temperature measuring device is disposed in the second box;
  • the heating device is disposed on an inner side of the first box door and a side portion of the second box body, specifically: six heating sheets, respectively disposed on the inner side of the first box door, on the second box body Side, lower side, rear side, right side and left side;
  • a fan heater 611 disposed in the enclosed space, such as an inner wall of the first case opposite the rear side of the second case;
  • a heat insulating device 67 disposed on an outer side portion of the first case or an inner side portion of the first case, such as an inner wall of the first case or an outer wall of the second case, such that the first Forming an enclosed space between the box and the second box;
  • the fan is disposed in a space formed by the second box;
  • the motor 68 controls the direction of rotation of the fan, and a reasonable wind direction causes the gas in the second tank to quickly reach uniformity.
  • the output power of the heating sheet is adjusted, and whether the heating or heating power is selected according to the corresponding wall surface, and the motor 68 is controlled to realize the automatic steering of the fan 610 to control the wind direction. Fine adjustment of temperature uniformity control in the cabinet 65; when there are extreme cases, when the heating sheet has multiple failures, in order to ensure the survival rate of the cultured cells, the second housing 65 is slowly delayed by reducing the output power of the available heating sheets. The internal temperature is reduced, and the motor 6.8 is adjusted again. The fan 610 is controlled to turn at a large angle, and the output power of the fan heater 611 is increased to achieve stable temperature control, and the cell culture work in which the second tank 65 has been placed is continuously performed until the end of the culture.
  • FIG. 3 is a schematic structural diagram of a cell culture workstation according to an embodiment of the present invention. As shown in FIG. 3, the cell culture workstation includes:
  • a cabinet 2 a console 4, a glove hole 5, a transparent plate 3 and a moving wheel 1, the glove hole being disposed on a front side portion of the cabinet;
  • the front side portion of the first tank body 62 is fixed on the rear side portion of the cabinet, and the rear side portion of the cabinet has the first The position and size of the door 61 are matched to the through holes.

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Abstract

本发明提供了一种细胞培养箱，包括：第一箱体；第二箱体，所述第二箱体设置在所述第一箱体内部，第二箱体和第一箱体之间形成封闭的空间；第一箱门，所述第一箱门设置在所述第一箱体的前侧部上，第一箱体和第一箱门之间具有密封件；测温器件，所述测温器件设置在所述第二箱体内；加热器件，所述加热器件设置在所述第一箱门的内侧以及第二箱体的侧部；风扇加热器，所述风扇加热器设置在所述封闭的空间内；隔热器件，所述隔热器件设置在所述第一箱体的外侧部或第一箱体的内侧部。本发明具有温控效果好等优点。

Description

细胞培养箱及其工作方法 技术领域
本发明涉及细胞培养，特别涉及细胞培养箱及其工作方法。
背景技术
在医学、生命健康、环境学等领域中，细胞培养工作站提供了超净密闭空间的培养环境(达到ISO 5或是GMP级)，而内置的培养箱是细胞在模拟体内生理环境培养的专用装置，应具有温度恒定、温度均一性高、温度恢复快速性、无温度差引起的冷凝水现象等温控特性。目前的培养箱加热方式有：
1.水套式，该加热方式的稳定性很大程度上取决于电热丝的稳定性与响应时间，即选择合理的电热丝厚度和直径来减缓电热丝元素的内部消耗，且需要不定时的对水箱进行加水、清洗、清空及监控，这对于培养箱设计变得尤为困难。
2.气套式，该加热方式存在受热均一性较差，内循环式的内腔壁增设的风道和风口对设计要求高、成本较高，外循环式的结构相对简单，存在导热性不均。
3.直热式，该加热方式温控精度高，六面加热方式可基本消除玻璃门的冷凝水问题，但会造成候箱体内的温度场分布不均匀，破坏细胞培养环境的一致性，且当培养箱内壁有一面及其以上出现故障时，对腔室的温度控制稳定性具有很大影响，尤其对加热片更换、零部件更换等对细胞培养环境的稳定性、连续性带来极大的难度。
发明内容
为解决上述现有技术方案中的不足，本发明提供了一种温控精确、温度均一性好的细胞培养箱。
本发明的目的是通过以下技术方案实现的：
一种细胞培养箱，所述细胞培养箱包括：
第一箱体；
第二箱体，所述第二箱体设置在所述第一箱体内部，第二箱体和第一箱体之间形成封闭的空间；
第一箱门，所述第一箱门设置在所述第一箱体的前侧部上，第一箱体和第一箱门之间具有密封件；
测温器件，所述测温器件设置在所述第二箱体内；
加热器件，所述加热器件设置在所述第一箱门的内侧以及第二箱体的侧部；
风扇加热器，所述风扇加热器设置在所述封闭的空间内；
隔热器件，所述隔热器件设置在所述第一箱体的外侧部或第一箱体的内侧部。
本发明的目的还在于提供了一种温控精度高、加热均匀的细胞培养工作站，该发明目的通过以下技术方案得以实现：
一种细胞培养工作站，所述细胞培养工作站包括机柜、操作台、手套孔，所述手套孔设置在所述机柜的前侧部上；所述细胞培养工作站进一步包括：
细胞培养箱，所述细胞培养箱采用上述的细胞培养箱，第一箱体的前侧部固定在所述机柜的后侧部上，所述机柜的后侧部具有与第一箱门的位置和大小匹配的通孔。
本发明的目的还在于提供了上述细胞培养箱的工作方法，该发明目的通过以下技术方案得以实现：
根据上述的细胞培养箱的工作方法，所述工作方法包括以下步骤：
(A1)关闭第一箱门，开启风扇加热器和加热器件，直到测温器件输出的温度达到设定值；
(A2)关闭风扇加热器，再关闭加热器件。
与现有技术相比，本发明具有的有益效果为：
1.培养箱内温控精度更高、准确、温度场分布均匀、培养一致性效果好；
2.气套加热恒温和包裹式直热方式有效解决透明箱门及气路管路的冷凝水、温控效率高，温度恢复快；
3.结构简单、紧凑。
附图说明
参照附图，本发明的公开内容将变得更易理解。本领域技术人员容易理解的是：这些附图仅仅用于举例说明本发明的技术方案，而并非意在对本发明的保护范围构成限制。图中：
图1是根据本发明实施例1的细胞培养箱的结构简图；
图2是根据本发明实施例1的细胞培养箱的A-A剖视图；
图3是根据本发明实施例2的细胞培养工作站的结构简图。
具体实施方式
图1-3和以下说明描述了本发明的可选实施方式以教导本领域技术人员如何实施和再现本发明。为了教导本发明技术方案，已简化或省略了一些常规方面。本领域技术人员应该理解源自这些实施方式的变型或替换将在本发明的范围内。本领域技术人员应该理解下述特征能够以各种方式组合以形成本发明的多个变型。由此，本发明并不局限于下述可选实施方式，而仅由权利要求和它们的等同物限定。
实施例1：
图1-2示意性地给出了本发明实施例的细胞培养箱的结构简图，如图1-2所示，所述细胞培养箱包括：
第一箱体62，如由抛光电解不锈钢材料制成；
第二箱体65，所述第二箱体设置在所述第一箱体内部，第二箱体和第一箱体之间形成封闭的空间；
第一箱门61，所述第一箱门设置在所述第一箱体的前侧部63上，第一箱体 和第一箱门之间具有密封件；
第二箱门64，所述第二箱门设置在所述第一箱体的前侧部上，且处于所述第一箱门的内侧，所述第二箱体门采用透明材料，如玻璃；所述第二箱门上具有至少二个通孔，以及封堵所述通孔的透明门642-643；
测温器件69，如温度传感器，所述测温器件设置在所述第二箱体内；
加热器件66，所述加热器件设置在所述第一箱门的内侧以及第二箱体的侧部，具体为：6个加热片，分别设置在第一箱门内侧，第二箱体的上侧、下侧、后侧、右侧和左侧；
风扇加热器611，所述风扇加热器设置在所述封闭的空间内，如与第二箱体的后侧相对的第一箱体的内壁上；
隔热器件67，所述隔热器件设置在所述第一箱体的外侧部或第一箱体的内侧部，如第一箱体的内壁上或第二箱体的外壁上，使得第一箱体和第二箱体之间形成隔热的封闭空间；
风扇610，所述风扇设置在所述第二箱体形成的空间内；
电机68，所述电机控制所述风扇的转动方向，合理的风向使得第二箱体内气体快速达到均一性。
本发明实施例的上述细胞培养箱的工作方法，包括以下步骤：
(A1)关闭第一箱门和第二箱门，使得第二箱体内形成一个密闭空间，开启加热片，同时运行风扇610和风扇加热器611，当第二箱体65内的2个温度传感器69测温值达到35℃时，再控制加热功率，使得温度缓慢升高至37℃；
(A2)先关闭风扇加热器6.11，然后关闭风扇610和6个加热片；
当出现第二箱体65内部空间加热不均时，调节加热片输出功率，控制对应壁面是否选择加热或加热功率大小，同时通过控制电机68，实现风扇610自动转向来控制风向，起到第二箱体65内温度均一性控制的微调作用；当出现极端情况，加热片出现多个故障时，为保证培养细胞的成活率，通过降低可用加热片输出功率，起到缓慢延迟第二箱体65内温度的降低作用，再调节电机6.8， 控制风扇610大角度转向，同时加大风扇加热器611的输出功率，实现温度的稳定控制，保证第二箱体65已放置的细胞培养工作的持续正常进行，直至培养结束。
实施例2：
图3示意性地给出了本发明实施例的细胞培养工作站的结构简图，如图3所示，所述细胞培养工作站包括：
机柜2、操作台4、手套孔5、透明板3和移动轮1，所述手套孔设置在所述机柜的前侧部上；
细胞培养箱，所述细胞培养箱采用实施例1中的细胞培养箱，第一箱体62的前侧部固定在所述机柜的后侧部上，所述机柜的后侧部具有与第一箱门61的位置和大小匹配的通孔。

Claims (9)

  1. 一种细胞培养箱，其特征在于：所述细胞培养箱包括：
    第一箱体；
    第二箱体，所述第二箱体设置在所述第一箱体内部，第二箱体和第一箱体之间形成封闭的空间；
    第一箱门，所述第一箱门设置在所述第一箱体的前侧部上，第一箱体和第一箱门之间具有密封件；
    测温器件，所述测温器件设置在所述第二箱体内；
    加热器件，所述加热器件设置在所述第一箱门的内侧以及第二箱体的侧部；
    风扇加热器，所述风扇加热器设置在所述封闭的空间内；
    隔热器件，所述隔热器件设置在所述第一箱体的外侧部或第一箱体的内侧部。
  2. 根据权利要求1所述的细胞培养箱，其特征在于：所述加热器件设置在所述第二箱体的外壁上，所述隔热器件设置在所述第一箱体的内壁上。
  3. 根据权利要求1所述的细胞培养箱，其特征在于：所述风扇加热器设置在所述第一箱体的与所述第二箱体的后侧部相对的后侧部的内壁上。
  4. 根据权利要求1所述的细胞培养箱，其特征在于：所述细胞培养箱进一步包括：
    风扇，所述风扇设置在所述第二箱体形成的空间内。
  5. 根据权利要求4所述的细胞培养箱，其特征在于：所述细胞培养箱进一步包括：
    电机，所述电机控制所述风扇的转动方向。
  6. 根据权利要求1所述的细胞培养箱，其特征在于：所述细胞培养箱进一步包括：
    第二箱门，所述第二箱门设置在所述第一箱体的前侧部上，且处于所述第 一箱门的内侧，所述第二箱体门采用透明材料。
  7. 根据权利要求6所述的细胞培养箱，其特征在于：所述第二箱门上具有至少二个通孔，以及封堵所述通孔的透明门。
  8. 一种细胞培养工作站，所述细胞培养工作站包括机柜、操作台、手套孔，所述手套孔设置在所述机柜的前侧部上；其特征在于：所述细胞培养工作站进一步包括：
    细胞培养箱，所述细胞培养箱采用权利要求1-7任一所述的细胞培养箱，第一箱体的前侧部固定在所述机柜的后侧部上，所述机柜的后侧部具有与第一箱门的位置和大小匹配的通孔。
  9. 根据权利要求1-7任一所述的细胞培养箱的工作方法，所述工作方法包括以下步骤：
    (A1)关闭第一箱门，开启风扇加热器和加热器件，直到测温器件输出的温度达到设定值；
    (A2)关闭风扇加热器，再关闭加热器件。
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