Primary Cell Culture

Primary Culture is the cell culture system that is formed by culture cells directly obtained from tissue. Primary cell culture is more strictly referred to as the culture before successful passage. At this point, the cell retains the basic properties of the original cell. In practice, the first to the tenth generation of cultured cells are generally referred to as primary cell culture. Primary culture is mainly tissue block culture and dispersed cell culture.

• Organization shearing

1. Clean the obtained tissue with D-Hanks or Hanks solution to remove blood stains on the surface.
2. After cleaning again, cut the tissue into several small pieces with a scalpel, add an appropriate amount of buffer, and cut the tissue repeatedly with elbow eye scissors until the tissue becomes a paste, about 1mm3 in size.
3. Let it sit for a while, remove the top liquid with a straw, add the appropriate buffer and clean again.

• Digestion and separation
  Digest and separate small tissue pieces into cell clusters or dispersed single cells for further culturing. Commonly used digestive enzymes are trypsin and collagenase.
• Cultivation

1. Count cell suspensions with a counting plate. Adjust the number of cells to (2-5) × 105 cells/ml with the culture medium, or the density required for the experiment, and distribute them in culture flasks.
2. Place cells in a CO2 incubator, 5% CO2, at 37°C. The primary culture cells can adhere to the bottle wall, and start to grow after 3-5 days.
3. Add the new medium with 1/2 of the original medium, and change the medium after culturing for 2-3 days. Generally, the bottle wall can be overgrown in 7-14 days for passage.

Fig. 1 The basic flow chart for primary cell culture.

The in vitro time of primary cultured cells is short, and the characters are similar to those in vivo, so it is suitable for research.

• Provide a powerful means to study the growth, metabolism and reproduction of organism cells, and create conditions for subsequent subculture.
• For various experiments, such as drug testing, cell differentiation and virology experiments.
• Used as in vitro tools for preclinical and investigative biological studies, such as the study of inter- and intracellular communication, developmental biology, and elucidation of disease mechanisms such as cancer, Parkinson's disease, and diabetes.

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.

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.

Tweezers, Scissors and Cutters

Scissors are hand-held instruments designed to cut thin materials, such as paper and cloth, consisting of two blades laid one on top of the other. They may be intended for use in specific applications such as cutting tissue or bone, dissections, etc. Scissors are used in the laboratory to cut various materials. Some are for general use and others are designed specifically for tissue dissection or surgery.

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.

Water Baths

A water bath is laboratory equipment made from a container filled with heated water. It is used to incubate samples in water at a constant temperature over a long period of time. Most water baths have a digital or an analogue interface to allow users to set a desired temperature. A laboratory water bath is a preferred heat source for heating flammable chemicals instead of an open flame to prevent ignition.

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.

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.

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.

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 primary cell culture, please feel free to contact us.