Smooth muscle cells of chicken pulmonary artery

Smooth muscle cells of chicken pulmonary artery

Product attributes:

Cell Introduction:

Chicken pulmonary artery fibrosis isolated from pulmonary artery tissue; Pulmonary artery, also known as pulmonary artery trunk. In vertebrates that breathe air, the arteries that direct venous blood from the heart to the lungs. The pulmonary artery originates from the right ventricle and runs obliquely to the left, upper, and rear before the aorta. It divides into the left and right pulmonary arteries below the aortic arch and enters the lungs through the pulmonary hilum. The pulmonary artery trunk is located inside the pericardium and is a thick and short arterial trunk. Originating from the right ventricle, it runs obliquely in the upper left posterior direction in front of the ascending aorta, and divides into the left and right pulmonary arteries below the aortic arch. The left pulmonary artery is relatively short and runs horizontally in front of the left main bronchus, dividing into two branches that enter the upper and lower lobes of the left lung. The right pulmonary artery is long and thick, running horizontally to the right after passing through the ascending aorta and superior vena cava, and branching into three branches at the right pulmonary hilum to enter the upper, middle, and lower lobes of the right lung. Fibroblast cells（Fibroblast）It is the main cellular component of loose connective tissue, differentiated from mesenchymal cells during embryonic development; Fibroblasts are relatively large, with clear contours and mostly flat structures resembling spindle or star shaped protrusions. Their nuclei are regular oval shaped, and their nucleoli are large and distinct. Fibroblasts have strong functional activity, weak alkaline cytoplasm, and significant protein synthesis and secretion activities. Under certain conditions, they can achieve interconversion with fibroblasts; Fibroblasts play a crucial role in the repair of varying degrees of cellular degeneration, necrosis, and tissue defects. The newly isolated pulmonary artery fibroblasts are round in shape and have good refractive index, suspended in culture medium.30minCells adhere to the wall, and some of them begin to protrude pseudopodia, manifested as small protrusions;6hThe posterior cells are basically adhered to the wall, stretched into a spindle shape, with clear nuclei, evenly distributed, scattered growth, and do not aggregate into clusters; Cells grow rapidly,5-7The sky is in a fused state, with tightly arranged cells, some overlapping and growing, flat, and larger cell bodies. The cytoplasm is transparent, and the nucleus is large, oval shaped, and light in color. Cells fuse and connect with each other to form a network; The cells are distributed in a flat spindle or star shape with protrusions. The main functions of pulmonary artery fibroblasts under physiological conditions include: constructing and maintaining the normal morphology of pulmonary arteries; Synthesize and release extracellular matrix; Promptly gather and repair damaged tissues in large quantities after organizational injury.

Method Introduction:

The chicken pulmonary artery smooth muscle isolated in the company's laboratory was prepared using collagenase digestion combined with differential adhesion method, with a total cell count of approximately5×10?cells/Bottle.

Quality inspection:

The smooth muscle of chicken pulmonary artery isolated from the company's laboratory was subjected to alpha massage-SMAImmunofluorescence 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