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Method and device for preparing lentivirus

A lentivirus and container technology, applied in the field of lentivirus preparation, can solve the problems of cell shedding, affecting the success rate of cell transfection, and not forming a closed state, so as to avoid cell detachment, reduce labor costs, and fully mix the effect

Pending Publication Date: 2021-09-28
CHONGQING PRECISION BIOTECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

It is directly connected to the cell culture container without any intermediate device for buffering and pausing, and the cell factory connector is only a single-channel connector, which can easily lead to the formation of a faster flow of the plasmid mixture. The flow will shed the cells adhered to the wall of the cell factory, seriously affecting the success rate of cell transfection, and the waste liquid tank device is not used in this technology, and the waste liquid needs to be collected manually every time, and the pollutants are easily released from the waste liquid pipe. into the entire preparation device, without forming a completely closed state

Method used

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  • Method and device for preparing lentivirus
  • Method and device for preparing lentivirus

Examples

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Embodiment 1

[0034] Example 1 A device for preparing lentivirus

[0035] figure 1 It is a schematic diagram of a device for preparing lentivirus. It should be noted that this device is only an example adapted to this application, and should not be considered as providing any limitation on the scope of use of this application. The device is also not to be construed as being dependent on or necessarily having figure 1 One or more components of the exemplary device for preparing lentivirus shown in .

[0036] like figure 1 As shown, the present invention discloses a device for preparing lentivirus, comprising a mixing container 1 and a cell culture container 3, the first pipeline 20 through which the mixing container 1 passes is connected to a liquid storage bag 2, and the liquid storage bag 2 passes through a flow-stopping The second pipeline 30 of the device is connected to the cell culture container 3 , and the cell culture container 3 is connected to the waste liquid container 4 throug...

Embodiment 2

[0050] Embodiment 2 A kind of method for preparing lentivirus

[0051] In the method of preparing lentivirus using the device in Example 1, the mixed transfection complex is pumped into the storage bag 2 containing the culture substrate, and the transfection complex mixed with the culture substrate is pumped into the cell culture device 3 Lentiviral culture in.

[0052] Based on the above method, the medium in the storage bag 2 is 2% DMEM, and the cell culture device 3 is at 37°C, 5% CO 2 The lentivirus was obtained by culturing for 40-48 hours under the above conditions. The waste liquid generated in the cell culture device 3 and the excess transfection complex mixed with the culture medium in the infusion bag can be discharged into the waste liquid container 4 .

[0053] The specific operation steps are as follows:

[0054] (1) Use the mixing container 1 with a specification of 2L and the cell factory connector 32 for use after sterilization, and adjust the size of the on...

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Abstract

The invention relates to the field of lentivirus preparation, in particular to a method and device for preparing a lentivirus. According to the device for preparing the lentivirus, a liquid storage bag holding a culture medium is additionally arranged, and the culture medium does not need to be pumped from a cell factory, so that a transfection compound and the culture medium are mixed in the liquid storage bag, and the mixture is further fully mixed, so that the operation efficiency is improved, and a certain buffering effect can be achieved. In addition, a waste liquid container is further arranged, so that a closed mixing device is formed, and the risk of environmental pollution is reduced.

Description

technical field [0001] The invention relates to the field of lentivirus preparation, in particular to a method and device for preparing lentivirus. Background technique [0002] Calcium phosphate method to prepare lentivirus, the more common way is to mix deionized water, calcium chloride, and plasmid in a container, then slowly add HBS solution inward, and at the same time mix it evenly by magnetic stirring to form a lentivirus containing Calcium phosphate-DNA complex, then add the transfection complex to the cell factory, mix the inner medium with the transfection complex by shaking the cell factory. Since the process of mixing transfection reagents to form a calcium phosphate-DNA complex is an open environment (inside a biological safety cabinet, the container is open), when adding materials such as plasmids, it is easy to introduce microorganisms and there is a risk of contamination. The transfection complex needs to be transferred into the cell factory and mixed with t...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12M3/00C12M1/24C12M1/12C12M1/04C12M1/02
CPCC12M23/08C12M23/14C12M23/38C12M23/58C12M27/02C12M29/04C12M33/04C12M33/14C12M37/04
Inventor 黄一杨周琴杨智张茜真沈俊杰晏波波郑姣李运燕王玲琳王戬
Owner CHONGQING PRECISION BIOTECH CO LTD
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