Porcine preadipocytes

Porcine preadipocytes

Product attributes:

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, ultimately becoming mature adipocytes. Pre adipocytes have strong resistance to external mechanical damage and are expected to become beneficial fillers for soft tissue defects.

Cultivation conditions for primary cells:

　　1、 Culture of adherent cells (including semi adherent cells) in a static state

1. Cells should be thoroughly rinsed to remove the toxicity of digestive juices as much as possible;

2. When inoculating cells, the concentration should be slightly higher, at least 5 × 108 cells/L;

3. The culture medium can be Eagle (MEM Corning 10-010-CVR) or DMEM (Corning 10-013-CVR) for cultivation;

4. The concentration of fetal bovine serum is 10% -80%

5. It should be cultured in a 37 ℃ 5% CO2 incubator;

6. Try to minimize oscillation in the first 2 days to prevent newly adhered cells from shedding and floating;

7. When the cells are basically attached to the wall and gradually form a network, the primary cells should be replaced with fluid;

8. When using suspended cells from bone marrow or peripheral blood for one week of static culture, the cell suspension should be centrifuged at low speed and then replaced.

　2、 Suspension cell culture

1. During primary culture, red blood cells should be removed as much as possible;

2. Short term cultivation can be carried out in RPMI1640 medium containing 10% calf serum;

3. Cell concentration can be within the range of 5-8 × 109/L for bottle separation experiments;

4. During long-term cultivation, growth factors should be added to lymphocytes, and a small amount of original patient serum should be added to leukemia cells to promote cell growth;

5. Cell fluid exchange usually requires a half volume exchange every 3 days;

6. Cell passage can only be carried out when cell proliferation accelerates and cell density is high.

Method Introduction:

The pre fat of pigs isolated in the company laboratory was prepared by collagenase digestion method, with a total cell count of approximately5×10?cells/Bottle.

Quality inspection:

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.

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%

Cell passage and cryopreservation:

There are generally four methods for culturing 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 focus on observing the connections, arrangements, and interactions between cells, as well as the biological regulatory effects of local environments.

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