Porcine preadipocytes

Primary cell culture conditions:

Temperature: Usually conducted at 37 ℃ to simulate the internal temperature.

PH value: The pH value of the culture medium should be maintained between 7.2 and 7.4 to maintain the normal physiological state of the cells.

Gas environment: Conducted in an incubator containing 5% CO2 to promote cell growth.

Culture medium: Use a medium containing appropriate nutrients, such as Eagle's MEM, DMEM, etc.

Aseptic operation: The entire cultivation process needs to be carried out under sterile conditions to prevent contamination.

This product is only for scientific experimental research use! Cannot be used for clinical or animal diagnosis!

Cell Introduction:

Porcine preadipolar separation from adipose tissue; Adipose tissue is mainly composed of a large number of clustered adipocytes, which are separated into lobules by thin layers of loose connective tissue; The stored fat can be quickly broken down into glycerol and fatty acids when needed, and transported to various tissues through the bloodstream for utilization. They affect insulin sensitivity, blood pressure levels, endothelial function, fibrinolytic activity, and inflammatory response, and are involved in various important pathophysiological processes; Adipose tissue has evolved from a simple energy storage organ in the past to an extremely important endocrine system. Adipose tissue contains two main types of cells in the body: mature adipocytes that accumulate lipid droplets in the cytoplasm; Another type is preadipocytes that do not accumulate lipid droplets in the cytoplasm but have this potential. Preadipocytes are spindle shaped and have the ability to divide and proliferate; Mature adipocytes are round in shape and have lost their ability to divide and proliferate. Due to the fact that preadipocytes are a specific type of precursor cell with the ability to proliferate and differentiate into adipocytes, they are closely related to obesity. Pre adipocytes are a type of fibroblast that continuously absorb lipids during their evolution and eventually become mature adipocytes. Pre adipocytes have strong resistance to external mechanical damage and are expected to become beneficial fillers for soft tissue defects.

Method Introduction:

The pre fat of pigs separated in the company's laboratory is treated with oil redOStaining detection, purity can reach90%Above, and not containingHIV-1TheHBVTheHCVMycoplasma, bacteria, yeast, fungi, etc.

Quality inspection:

The pre fat of pigs isolated in the company laboratory was prepared by collagenase digestion method, with a total cell count of approximately5×105cells/Bottle.

Training Information:

The culture medium containsFBSGrowth additivesPenicillinTheStreptomycinwait

Fluid change frequency per2-3Change the fluid once every day

Growth characteristics: Wall adhesion

Cell morphology spindle shaped, polygonal

Passage characteristics can be passed down3Dai Zuo

digestive juice0.25%

Cultivation conditions gas phase: air,95%；CO2，5%

The specific steps for primary cell culture are as follows:

Organizational processing: Remove animal tissue from the body, cut it into pieces, and treat it with digestive enzymes to disperse it into individual cells.

Cell counting: Use a blood cell counter or other methods to count cells to ensure appropriate cell density.

Inoculation culture: Inoculate the cell suspension into a culture bottle, add an appropriate amount of culture medium, and incubate in a 37 ℃, 5% CO2 incubator.

Liquid exchange and passage: Regularly exchange the culture medium, and carry out passage culture when the cells reach a certain density to maintain the growth state of the cells.

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Four cultivation methods for primary cells:

　　① Organizational block cultivation method

Tissue block culture is a commonly used, simple, and highly successful primary culture method. The basic method is to inoculate the cut small tissue clusters into a culture bottle (or dish), and the bottle wall can be pre coated with a thin layer of collagen to facilitate the adhesion of the tissue blocks to the bottle wall, allowing surrounding cells to grow outward along the bottle wall.

　　② Digestive cultivation method

　③ Suspension cell culture method

For cells that grow in suspension, such as leukemia cells, lymphocytes, bone marrow cells, cancer cells and immune cells in pleural and ascites, digestion is not required. They can be isolated by low-speed centrifugation and cultured directly, or cultured by inoculating after lymphocyte stratification.

　④ Organ culture

Organ culture refers to the direct cultivation of organs or tissue blocks obtained from donors under specific environmental conditions outside the body without tissue separation. Organ culture can maintain the relative integrity of organ tissues and can be used to observe the connections, arrangements, and interactions between cells, as well as the biological regulatory effects of local environments