How to do cell subculture? What are the necessary experimental consumables?

in cell experiments

Passage culture plays a key role

Its success or failure will be directly related to the experimental results.

Reliable consumables ensure experimental success

has a vital position

Aijin provides a full set of cell passaging experiment consumables

Will help you easily handle every step of cell passaging

From preparation stage to cultivation process

Comprehensively guarantee the smooth progress of your cell experiments

Chapter 1

Principles of cell subculture

Cells are constantly dividing in a culture container. When adherent cells grow into a dense monolayer and suspended cells fill the entire volume of culture medium, cell growth slows down or stops due to contact inhibition due to mutual contact between cells, depletion of nutrients in the culture medium and accumulation of cell metabolites.

At this time, the cells need to be diluted and aliquoted into new culture containers to expand the culture, that is, cell passage. Suspension cells can be divided into bottles directly, while adherent cells need to be digested and divided into bottles.

Chapter 2

Key consumables needed

During the entire cell subculture process, different experimental consumables are required at different stages. We can provide a variety of key consumables to help the experiment proceed smoothly.

Cell culture dish/bottle/plate

Aijin cell culture bottles/dishes/plates are made of imported medical-grade materials with high transparency, which can provide a good environment and observation conditions for cell growth, ensuring the visualization effect and growth stability of cells.

Serological pipette

Used for transferring trypsin, culture medium and other liquids and pipetting and mixing of cells. The clear scale on the surface of Aijin pipette facilitates accurate reading of liquids, which can help experimenters complete liquid operations efficiently and reduce errors.

centrifuge tube

For cell centrifugation operations, 15mL centrifuge tubes are commonly used. Aijin centrifuge tubes are available in sterile packaging, have good chemical resistance, are pyrogen-free, endotoxin-free, and DNase/RNase-free, ensuring no contamination during cell processing and stable centrifugation results.

Triangle culture shake flask

Suitable for the culture of suspension cells. Aijin triangular culture shake flasks are individually sterilely packaged and have excellent durability and sterility, and can meet the special culture needs of suspended cells.

Chapter 3

Specific steps for cell subculture

①Preparation stage

Preheat the culture medium (such as DMEM containing 10% fetal bovine serum), PBS buffer, trypsin and other reagents, and disinfect the experimental environment, equipment and consumables.

② Trypsin digestion

For adherent cells, generally after the cell density reaches 80%-90%, use 0.25% trypsin for digestion at 37°C. Observe under an inverted microscope. When most of the cells become round and fall off, and the cells move like quicksand, complete culture medium can be added to terminate the digestion.

③ Disperse cells by pipetting

Use a pipette or Pasteur pipette to repeatedly pipette the digested cells into a suspension until most of them have been blown into single cells under a microscope. Aijin pipettes have smooth walls, which can effectively reduce liquid adhesion, avoid bubbles, improve sampling accuracy, and lay the foundation for subsequent operations.

④Centrifugation

Transfer the cell suspension to a centrifuge tube and centrifuge at 1000 rpm/min for 5 minutes. Remove the supernatant and add new culture medium to resuspend the cells.

⑤Dilution and packaging

Dispense the cell suspension into new culture bottles, usually at a ratio of 1:3, and then add the remaining culture medium according to the volume of the culture bottle.

⑥Continue to cultivate

Place the new culture bottle ₂in a 37°C, 5% CO incubator to continue culturing. Generally, it will continue to be passaged after it reaches full growth in 2-3 days.

In short, cell subculture is a delicate process. Aijin provides cell experiment consumable solutions, covering a series of products such as cell culture bottles, pipettes, centrifuge tubes, etc., which can effectively improve experimental efficiency and ensure healthy cell growth. We hope to meet your consumable needs in cell subculture to the greatest extent!