Cell Subculture

Subculture (or passage) refers to the transfer of cells from one culture vessel to another culture vessel. Subculture usually (not always) involves the subdivision of proliferating cells that enables the propagation of a cell line.

Take a T25 flask as an example

• Passaging of suspension cells

1. Collect cells with medium and briefly centrifuge (150–300 ×g, 3–5 min) to pellet cells.
2. Remove the medium and gently resuspend cells with a balanced salt solution. Briefly centrifuge (150–300 ×g, 3–5 min) to pellet cells.
3. Remove salt solution, resuspend cells in fresh medium and count cells.
4. Plate cells onto a new dish at the desired cell density.

• Passaging of adherent cells

1. Remove medium.
2. Wash cells with calcium- and magnesium-free balanced salt solution (phosphate buffered saline, Hanks’ balanced salt solution, etc.)
3. Add detaching agent. Incubate at 37°C until cells are fully detached from the dish (2–20 min depending on the cell line).
4. Resuspend cells in fresh medium, pipette thoroughly to obtain single cell suspension. Count cells.
5. Place cells onto a new dish at the desired cell density

• Passaging of semi-adherent semi-suspended cells

1. Pour out the culture medium in the bottle, add sterile PBS and rinse once, and then add 2mL digestive fluid. Let it stand in the incubator for 3 minutes.
2. Observe the digested cells under an inverted microscope. If the cells become round and are no longer connected to each other into pieces, immediately add an equal proportion of fresh culture medium, blow, make the cell suspension and centrifuge.
3. Discard the supernatant, resuspend with 5ml culture medium, passage in a ratio of 1:3 (depending on the number of cells), and send back to the carbon dioxide incubator for further culture.

Fig. 1 Basic subculturing procedure.

1. Passage of cells allows the expansion of cultured cells (formation of cell lines), cell cloning, easy to preserve, but may lose some special cellular and differentiation characteristics.
2. The greatest benefit of cell line formation from passage cells is that it provides a large amount of durable experimental material, which is convenient for experiments.

Ultra-clean Bench

The ultra-clean workbench is a kind of unidirectional flow air purification equipment that provides a local dust-free and sterile working environment. Most of the ultra-clean workbenches are 100 grades, which are mainly divided into two categories, one is horizontal laminar flow, and the other is vertical laminar flow. It can be used in pharmaceutical, medical and health, university scientific research laboratories and other fields.

Pipette

Pipette is a small piece of apparatus which typically consists of a narrow tube into which fluid is drawn by suction (as for dispensing or measurement) and retained by closing the upper end. Pipettes are a common laboratory tool and come in a multitude of channels, displacements and construction materials. There are different pipettes for all kinds of uses within the lab.

Colony Counter

Biological procedures often rely on an accurate count of bacterial colonies and cells. Colony counters are used to estimate a liquid culture’s density of microorganisms by counting individual colonies on an agar plate, slide, mini gel, or Petri dish. Typical applications include Ames testing, bacterial mutation assays, and E. coli bacterial colonies. The counting can be accomplished manually, or can be semi- or fully automatic.

Biological Microscope

Biological microscopes are used to study organisms and their vital processes. Microscopes used in this field range widely, from relatively simple optical microscopes to very advanced imaging systems used in cell research, forensic medicine, and state-of-the-art high resolution molecular studies. Biological microscopes can be one of many types of technologies. The most common biological microscopes are compound microscopes, inverted microscopes and stereomicroscopes.

Laboratory Centrifuges

A centrifuge is any device that applies a sustained centrifugal force—that is, a force due to rotation. The widest use of centrifuges is for the concentration and purification of materials in suspension or dissolved in fluids. Suspended particles denser than the suspending liquid tend to migrate toward the periphery, while those less dense move toward the centre.

Laboratory Incubators

An incubator is a device used to grow and maintain microbiological cultures or cell cultures. It is made up of a chamber with a regulated temperature. An incubator maintains optimal temperature, humidity, and other conditions such as the CO₂ and oxygen content of the atmosphere inside. Incubators are essential for much experimental work in cell biology, microbiology, and molecular biology.

pH Meters and Osmometers

A pH meter is a scientific instrument that measures the hydrogen-ion activity in water-based solutions, indicating its acidity or alkalinity expressed as pH. The pH meter measures the difference in electrical potential between a pH electrode and a reference electrode. The pH meter is used in many applications ranging from laboratory experimentation to quality control.

STEMart provides you with a variety of cell culture equipment or consumables to meet your various R&D and application needs. If you have any questions or requirements for cell subculture, please feel free to contact us.