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Micro-scale cell electro-transfection micro-fluidic chip, electro-transfection sorter and applications thereof

A microfluidic chip, micro cell technology, applied in microorganisms, biochemical instruments, electric/wave energy processing enzymes, etc., can solve the problems of difficult cell purification, inconsistent transfection voltage frequency, large number of plasmids, etc. Quality, ease of cell culture, and the effect of simplified procedures

Pending Publication Date: 2018-05-04
ALLIFE MEDICAL SCI & TECH CO LTD
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  • Summary
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  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In the current preparation process of IPS induced pluripotent stem cells, among the transfection methods used, the transfection vectors used are mainly viruses, RNA, plasmids, etc., and the transfection methods used are mainly electroporation transfection method. The electroporation transfection method has the disadvantage of low transfection efficiency. During the transfection process, most of the cells cannot be transfected due to uneven voltage, resulting in waste, and the transfected cells and untransfected cells exist at the same time. The purification of cells Causes difficulties; in order to ensure the base number of transfected cells requires a very large number of cells (more than 1 million), the number of plasmids that need to be used is extremely large, resulting in a large amount of plasmids being wasted, which inevitably leads to increased costs, so a kind of A new IPS cell transfection culture method for cell transfection
[0004] The main problems in the prior art are as follows: 1. The transfection voltage and frequency are not uniform in the existing electroporation transfection method, which causes differences in the conditions of cell transfection, resulting in differences in the quality of transfected cells; 2. The electrotransfection voltage control of the existing transfection method is troublesome and the electrotransfer efficiency is low; 3. In order to ensure that enough transfected adult cells are finally obtained in the existing electrotransfected cells, it is necessary to use at least 1 million adult cells and consume a large amount of plasmids, and the cost is unreasonable. Avoid rising; 4. Adult cells need to be purified and separated after transfection, and clone culture. In the existing method, transfected cells and untransfected cells exist at the same time, which is difficult for cell purification and consumes a lot of manpower and material resources for later cell culture.

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  • Micro-scale cell electro-transfection micro-fluidic chip, electro-transfection sorter and applications thereof

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Embodiment Construction

[0048] The following is attached Figure 1-8 The present invention is described in further detail.

[0049] A microfluidic chip for microcell electroporation according to the present invention, comprising: an inlet A for adding cells to be transfected and electroporation buffer, and an outlet B for finally flowing out the transfected cells, which can apply negative pressure (Negative pressure) Negative pressure channel C for adsorbing and fixing cells, positive pressure (Positive pressure) can be applied for positive pressure channel D for inputting transfection vector plasmids, and main channel E for normal cell flow and completion of electroporation process ;

[0050] The micro-cell electrotransfer microfluidic chip 7 includes upper and lower layers, and the main channel E between the upper and lower two layers: the upper layer of the micro-cell electrotransfer microfluidic chip 7 is a flat plate for connecting electrodes, Voltage can be applied; there are 96 grooves in th...

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Abstract

The invention relates to a micro-scale cell electro-transfection micro-fluidic chip, an electro-transfection sorter and applications thereof. The micro-scale cell electro-transfection sorter comprisesan electro-transfection unit, a display screen, an exterior box, a power supply unit, a micro control unit, and a main sensor. The electro-transfection unit comprises the chip. The display screen isused to send a command to the micro control unit and receive and display feedback information from the micro control unit and the main sensor. The micro control unit is used to receive the command sent by the display screen and control the electro-transfection unit and the power supply unit. The electro-transfection unit is used to complete cell transfection. The main sensor is used to receive feedback information from the electro-transfection unit and send the feedback information to the display screen and the micro control unit. The micro-scale cell electro-transfection micro-fluidic chip comprises an injection port, a discharge port, a negative pressure channel, a positive pressure channel, and a main channel, and a 96-well plate is arranged behind the discharge port. The provided chipcan guarantee that the transfection states in the main channel are identical during the transfection process, the transfection efficiency is guaranteed, the cell quality is guaranteed by the 96-well plate, and the subsequent cell culture becomes more convenient.

Description

technical field [0001] The invention relates to the field of IPS induced pluripotent stem cells, stem cell induced differentiation, specifically a micro-cell electroporation microfluidic chip, electroporation sorting apparatus and application thereof. Background technique [0002] Through the induction of exogenous transcriptional regulatory factors, adult cells are reprogrammed into embryonic stem cell-like pluripotent cells, which are called induced pluripotent stem cells (iPS cells), and this method is called iPS technology. The birth of iPS technology has pushed stem cell research to a new level and greatly enriched the content of stem cell research. In the past 3 years, iPS technology has developed rapidly, and has achieved success in mice, humans, macaques, rats and pigs. In particular, the research on iPS in mice and humans has been deepening, and the induction method has been continuously optimized. Various cells in animal tissues can be induced into iPS cells. Sin...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12M1/42C12M1/36C12M1/34C12N5/00C12N13/00
CPCC12M23/16C12M35/02C12M41/00C12N5/00C12N13/00
Inventor 顾春雨张会远
Owner ALLIFE MEDICAL SCI & TECH CO LTD
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