CN-121975618-A - Stromal cell culture device and control method thereof

CN121975618ACN 121975618 ACN121975618 ACN 121975618ACN-121975618-A

Abstract

The invention discloses a matrix cell culture device and a control method thereof, which belong to the technical field of cell culture devices and comprise a main cavity of an incubator, a micro-transfer cabin and a drawer type tray, wherein the inside of the main cavity of the incubator is maintained in a constant temperature, high humidity and positive pressure state relative to the external environment, the micro-transfer cabin is arranged on the front wall of the main cavity of the incubator and forms an independent and sealable transition cavity, the micro-transfer cabin is selectively communicated with the main cavity of the incubator through an openable gate valve at the inner side of the micro-transfer cabin and is separated from the external environment through an sealable outer door at the outer side of the micro-transfer cabin, the drawer type tray for bearing a culture dish is arranged in the micro-transfer cabin, and when the micro-transfer cabin is operated, the outer door is closed, the micro-transfer cabin is purified, and then the gate valve is opened, the drawer type tray is pushed into the main cavity of the incubator, so that the safe transfer of the culture dish is completed. According to the substrate cell culture device, the in-out operation of the culture dish is completed on the premise that the main cavity door is not required to be opened manually, so that pollution risks and environmental disturbance are obviously reduced.

Inventors

XU ZHIGUO
LIU CHAO
NIU YIKUN
REN ZHENHUI
YANG XUWEI

Assignees

湖州学院
协和华东干细胞基因工程有限公司

Dates

Publication Date: 20260505
Application Date: 20260116

Claims (10)

1. A stromal cell culture device comprising: the main cavity of the incubator is internally maintained in a constant-temperature, high-humidity and positive-pressure state relative to the external environment; The micro-transfer cabin is arranged on the front wall of the main cavity of the incubator and forms an independent sealable transition cavity, is selectively communicated with the main cavity of the incubator through an openable gate valve at the inner side of the micro-transfer cabin and is separated from the external environment through a sealable outer door at the outer side of the micro-transfer cabin, and is used for realizing the transition between the external environment of the culture dish and the main cavity of the incubator; the pushing mechanism comprises a screw rod assembly driven by a motor and is used for pushing the drawer type tray or the sliding rail bearing platform into or out of the main cavity; The mechanical arm system is arranged at the top or the side wall of the main cavity of the incubator, is a rectangular coordinate mechanical arm or a SCARA mechanical arm with more than three degrees of freedom, is provided with an arm body structure capable of moving along the X, Y, Z direction, and the tail end of the mechanical arm system is connected with a pneumatic clamping jaw type end effector for clamping a culture dish; When the drawer type tray or the slide rail bearing platform completely enters the main cavity of the incubator, the pneumatic clamping jaw type end effector clamps stacked culture dishes from the bearing partition plate in the main cavity of the incubator and places the culture dishes in the drawer type tray or the slide rail bearing platform or places the culture dishes in the drawer type tray or the slide rail bearing platform back into the main cavity of the incubator; the environment regulation and control module comprises a vacuum pump, and is connected to a bottom extraction opening of the micro-transfer cabin through a first electromagnetic valve and a first pipeline; The degerming and filtering gas recharging passage comprises a compressed gas source, a particle filter, a degerming filter and a second electromagnetic valve, and is connected to the top air inlet of the micro-transfer cabin through a second pipeline; The sterilization unit is an ultraviolet lamp or a hydrogen peroxide spray generator which is arranged on the inner wall of the micro-transmission cabin; The pressure sensor is arranged in the micro-transmission cabin and is used for monitoring the pressure in the cabin; The control unit is electrically connected with the vacuum pump, the electromagnetic valve, the sterilization unit and the pressure sensor, is used for executing a negative pressure pumping, filtering gas recharging and sterilization cycle on the interior of the micro-transfer cabin after the outer door is closed before the gate valve is opened, and is configured to execute the following operations: starting a first electromagnetic valve, starting a vacuum pump, and pumping the micro-transfer cabin to negative pressure of-30 kPa to-50 kPa; closing the first electromagnetic valve, opening the second electromagnetic valve, and filling clean air or nitrogen filtered by sterilization back into the cabin to normal pressure; starting a sterilization unit, and carrying out ultraviolet irradiation for more than or equal to 15 minutes or spraying atomized hydrogen peroxide and maintaining the concentration for more than or equal to 50ppm for 10 minutes; repeating the steps 1-3 for one time or multiple times to form a purification cycle; And the central control system is electrically connected with and coordinately controls the actions of the mechanical arm system, the propelling mechanism, the gate valve, the outer door and the environment regulation and control module, so that full-automatic sterile transmission in the process of taking and placing the culture dish is realized.
2. The stromal cell culture device of claim 1, wherein the sterilization unit is an ultraviolet lamp or a vaporized hydrogen peroxide generator, the ultraviolet lamp is disposed on the inner wall of the micro-delivery chamber, or the vaporized hydrogen peroxide generator is connected to the interior of the micro-delivery chamber by a conduit.
3. The stromal cell culture apparatus of claim 1, wherein the gate valve is controlled by a stepper motor or an electromagnetic drive device and is opened only when the drawer-type tray or the slide rail carrying platform is in or out, and the opening is provided that the pressure difference between the interior pressure of the micro-delivery cabin and the pressure of the main cavity is less than-50 Pa.
4. The stromal cell culture device of claim 1, wherein the pneumatically-actuated jaw end effector is a switchable dual-mode structure comprising: The vacuum sucker is used for adsorbing the bottom of the culture dish; The electric clamping jaw is used for clamping the edge of the culture dish; The mechanical arm is used for moving, and the mechanical arm system is provided with a force sensor and used for monitoring grabbing force in real time and preventing the culture dish from being damaged.
5. The stromal cell culture apparatus of claim 1, wherein a temperature and humidity sensor and a pressure sensor are disposed in the micro-transfer chamber, and the central control system dynamically determines whether the opening condition of the gate valve is satisfied according to sensor feedback, and records the environmental parameter change curve of each operation.
6. The stromal cell culture device of claim 5, wherein the central control system is equipped with a graphical user interface that supports user-defined robotic arm motion paths, sets sterilization parameters, views operation logs, and communicates with the laboratory information management system via a network interface.
7. The stromal cell culture device of claim 1, wherein the outer door of the micro-transport pod is an electrically operated door that is controlled by the central control system to open and close, and wherein the environmental conditioning module initiates a sterilization cycle when the outer door is closed and sealed.
8. The stromal cell culture device of claim 1, wherein the incubator main chamber is internally provided with a plurality of vertically distributed bearing partitions, and a storage layer area for placing culture dishes is formed between adjacent bearing partitions; And a plurality of vent holes are formed in the bearing partition plate so as to realize gas circulation among all storage layer areas and maintain uniform distribution of temperature and humidity in the main cavity of the incubator.
9. The stromal cell culture apparatus of claim 1, wherein the gate valve is a stainless steel plate with the thickness of 2mm, a plastic plate which is glued with the front wall of the main chamber body of the incubator is arranged at the periphery of the inlet of the drawer type tray or the slide rail bearing platform into the front wall of the main chamber body of the incubator, a slot which is anastomotic in size with the gate valve is arranged on the plastic plate from the upper edge to the lower edge, a notch which is anastomotic in size with the slide rail is arranged on the bottom edge of the gate valve, a groove is milled in the notch, an inflatable hollow silica gel O-shaped sealing ring is arranged in the groove, and after the gate valve is closed, the hollow silica gel O-shaped sealing ring is extruded and the slide rail is sealed.
10. A method of controlling a stromal cell culture device of claim 1, comprising the steps of: The extraction process comprises the following steps: T1, the mechanical arm moves to grab a culture dish in the main cavity of the incubator, and moves to the position above the drawer type tray or the sliding rail bearing platform after grabbing, and is slowly put down; t2, opening the gate valve, and pulling the drawer type tray or the whole slide rail bearing platform out of the main cavity of the incubator to the micro-transfer cabin through the pushing mechanism; t3, closing the gate valve; t4, opening an outer door, and manually taking out the culture dish from the drawer type tray or the sliding rail bearing platform; After T5 is completed, closing the outer door, and optionally starting the micro-transfer cabin to sterilize; The putting process comprises the following steps: S1, opening the outer door, and placing an external culture dish into the drawer type tray or the sliding rail bearing platform in the micro-transfer cabin by using a mechanical arm; s2, closing the outer door, and starting the environment regulation module to execute a negative pressure pumping, recharging and sterilizing cycle; s3, opening the gate valve, and pushing the drawer type tray or the sliding rail bearing platform into the main cavity of the incubator through the pushing mechanism; S4, closing the gate valve to finish putting in; s5, the mechanical arm moves to the upper portion of the drawer type tray or the sliding rail bearing platform, a culture dish in the drawer type tray or the sliding rail bearing platform is grabbed, and the mechanical arm moves to a placing position in the main cavity of the incubator.

Description

Stromal cell culture device and control method thereof Technical Field The invention relates to the technical field of cell culture devices in the technical field of biological experiment automation equipment, in particular to a matrix cell culture device and a control method thereof. Background In the cell culture process, the main cavity must be maintained at constant temperature (such as 37 ℃) for a long time, high humidity (> 95% RH), strict cleanliness, and positive pressure environment relative to the outside, and external air backflow prevention. If the main cavity door is directly opened to put in/take out the culture dish, the temperature and humidity balance can be destroyed, the risk of introducing external microorganism pollution can be destroyed, and the positive pressure state can be destroyed. When the door is opened, the mixture of the inside and outside air causes sudden temperature drop and humidity drop, and condensation or medium evaporation can be caused. Frequent opening and closing of the incubator door can cause severe fluctuation of temperature and humidity in the incubator and introduce risk of external microorganism pollution. The traditional manual operation mode has the problems of low efficiency, poor consistency, high pollution probability and the like. In the existing part of automatic systems, although a mechanical arm is adopted for transferring the culture dish, the environment isolation and dynamic sterilization capability of a transfer path are lacked, and the sterile transmission target of no interference in opening the door cannot be truly realized. Therefore, a full-automatic culture dish transmission system integrating automatic transmission, environmental isolation, dynamic sterilization and pressure balance control is needed, so as to solve the problems of environmental disturbance and cross contamination caused by human intervention in the prior art. Disclosure of Invention In order to overcome the defects in the prior art, one of the purposes of the invention is to provide a matrix cell culture device, which can finish the in-out operation of a culture dish without manually opening a main cavity door, thereby remarkably reducing pollution risk and environmental disturbance. The second objective of the present invention is to provide a control method for a stromal cell culture apparatus, which can implement intelligent dish taking and placing operations. One of the purposes of the invention is realized by adopting the following technical scheme: A stromal cell culture device comprising: the main cavity of the incubator is internally maintained in a constant-temperature, high-humidity and positive-pressure state relative to the external environment; The micro-transfer cabin is arranged on the front wall of the main cavity of the incubator, forms an independent sealable transition cavity, is selectively communicated with the main cavity of the incubator through an openable gate valve at the inner side of the micro-transfer cabin and is separated from the external environment through a sealable outer door at the outer side of the micro-transfer cabin, and is used for realizing the transition between the external environment of the culture dish and the main cavity of the incubator; the pushing mechanism comprises a screw rod assembly driven by a motor and is used for pushing the drawer type tray or the sliding rail bearing platform into or out of the main cavity; The mechanical arm system is arranged at the top or on the side wall of the main cavity of the incubator, is a rectangular coordinate mechanical arm or a SCARA mechanical arm with more than three degrees of freedom, is provided with an arm body structure capable of moving along the X, Y, Z direction, and the tail end of the mechanical arm system is connected with a pneumatic clamping jaw type end effector for clamping a culture dish; When the drawer type tray or the slide rail bearing platform completely enters the main cavity of the incubator, the pneumatic clamping jaw type end effector clamps stacked culture dishes from the bearing partition plate in the main cavity of the incubator and places the culture dishes in the drawer type tray or the slide rail bearing platform or places the culture dishes in the drawer type tray or the slide rail bearing platform back into the main cavity of the incubator; the environment regulation and control module comprises a vacuum pump, and is connected to a bottom extraction opening of the micro-transfer cabin through a first electromagnetic valve and a first pipeline; A degerming filtered gas recharging path comprising a compressed gas source, a particulate filter, a degerming filter (0.22 μm) and a second solenoid valve, connected to a top gas inlet of the micro transfer chamber through a second pipeline; The sterilization unit is an ultraviolet lamp or a hydrogen peroxide spray generator which is arranged on the inner wall of the micro-transmission cabin; The pressure s