Cell Cryopreservation

The principle of cell cryopreservation is "slow freezing", that is, cells are gradually frozen under the condition of slow cooling.

If the cell suspension is directly frozen at ultra-low temperature, the cells will suffer freezing damage and die. Protectants (e.g., DMSO, glycerol) added to the cryopreserved solution and isopropyl alcohol added to the cryopreserved box play a programmed cooling role.

• Use program freeze box

1. Prepare the program freeze solution and put it at room temperature for use or at 4℃ for pre-cooling.
2. Collect cells in logarithmic growth phase by centrifugation (adherent cells were digested and centrifuged, and suspended cells were centrifuged directly).
3. Resuspend cells with the prepared frozen storage solution, divide into the frozen storage tube according to 1-1.5 mL/ tube, tighten the tube cover, and make the mark.
4. Put the cryopreservation tube into the cryopreservation box, and put the cryopreservation box into the refrigerator at -80℃ overnight (24h).
5. Remove the freezing tube from the freezing box and transfer it to liquid nitrogen for long-term storage.

Fig. 1 Schematic diagram of using freezer container.

• Use serum-free non-programmed cryopreservation solution

1. Take out the serum-free non-programmed frozen solution from the refrigerator and set aside for later use.
2. Collect cells in logarithmic growth phase by centrifugation (adherent cells were digested and centrifuged, and suspended cells were centrifuged directly).
3. Discard the supernatant, add appropriate amount of serum-free non-programmed freezing solution, and resuspend the cells.
4. Divide into the frozen pipe according to the amount of 1-1.5mL/ tube, tighten the pipe cover, and make a mark.
5. Transfer the frozen storage tube directly to -80℃ refrigerator, and transfer it to liquid nitrogen for long-term storage after 24h.

Fig. 2 Schematic diagram of using serum-free non-programmed cryopreservation solution.

• Manual gradient cooling

1. Prepare the program freeze solution and put it at room temperature for use or at 4℃ for pre-cooling.
2. Collect cells in logarithmic growth phase by centrifugation (adherent cells were digested and centrifuged, and suspended cells were centrifuged directly).
3. Resuspend the cells with the prepared frozen storage solution, divided into the frozen storage tube according to 1-1.5 mL/ tube, tighten the tube cover, and make the mark.
4. Place the frozen storage tube in the order of [4℃ (20min) →-20℃ (30min) →-80℃ (24h) → liquid nitrogen].

Fig. 3 Schematic diagram of manual gradient cooling operation.

• Preserve the cellular properties, and revive cells for experiments when needed.
• Preserve cells, and prevent cells from being lost due to contamination of the cells being cultured or other unexpected events.

1. Cryopreservation of cells or organs; cryosurgery; biochemistry and molecular biology; food sciences; ecology and plant physiology.
2. Medical applications, such as blood transfusion, bone marrow transplantation, artificial insemination, and in vitro fertilization (IVF).
3. The possible banking of cells for human leukocyte antigen typing for organ transplantation, the allowance of sufficient time for transport of cells and tissues among different medical centers, and the provision of research sources for identifying unknown transmissible diseases or pathogens.
4. The long-term storage of stem cells is still the initial step toward tissue engineering, which holds promise for the regeneration of soft tissue esthetic function and for the treatment of known diseases that have currently no therapy option.

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.

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.

Liquid Nitrogen Container

Liquid nitrogen storage equipment is used to store biologic, genomic, and diagnostic samples in liquid nitrogen. Liquid nitrogen tanks can generally be divided into storage and transport tanks. The storage tank is mainly used for static storage of indoor liquid nitrogen, and it is not suitable for long-distance transportation under working conditions. The transportation tank has a special shockproof design to meet the transportation conditions.

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.

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.

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 Refrigeration

Laboratory refrigerators are designed with special features and options for use in the laboratory, including a range of refrigerator and freezer accessories. The primary function of a laboratory refrigerator is to maintain a defined, internal storage temperature in order to provide secure storage and protection of temperature-sensitive products, samples, specimens, chemicals, drugs, solutions, and other substances.

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 cryopreservation, please feel free to contact us.