Porcine adipose derived mesenchymal stem cells

Porcine adipose derived mesenchymal stem cells

Product attributes:

Cell Introduction:

Separation of porcine adipose derived mesenchymal stem 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. There are also some stem cell populations in adipose tissue that have self-renewal ability and multi-directional differentiation potential, known as adipose derived mesenchymal stem cells, abbreviated as adipose derived mesenchymal stem cells. Compared with bone marrow mesenchymal stem cells, they are highly similar in terms of source, cell population characteristics, and differentiation potential. However, adipose derived mesenchymal stem cells are more likely to obtain a sufficient number of cells and cause less damage to the body during the acquisition process, making them a more ideal source of autologous stem cells. Adipose tissue was isolated to obtain adipose derived mesenchymal stem cells with spindle shaped morphology as the main form,BrdUCan label its nucleus.

Method Introduction:

The porcine adipose derived mesenchymal stem isolated in the company's laboratory was prepared using collagenase digestion method, with a total cell count of approximately5×10?cells/Bottle.

Quality inspection:

Porcine adipose derived mesenchymal stem cells isolated in the company laboratoryCD90Immunofluorescence identification, 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 fibroblast like

Passage characteristics can be passed down5Dai Zuo;3Excellent condition within the generation

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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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