Pig heart microvascular endothelial cells

Primary cells vs. cell lines:

Cells directly isolated from human or animal tissues using enzymes or mechanical methods. Strictly speaking, cells isolated from the body before passage are called primary cells, and the vast majority of primary cells can divide 10-15 times in vitro (which is related to the length of telomeres). In addition, factors such as sampling and cost usually refer to cultured cells within the first to tenth generations as primary cells.

‍Pig heart microvascular endothelial cells

Product attributes:

Cell Introduction:

Separation of microvascular endothelium from pig heart tissue; The heart is an important organ in the vertebrate body, primarily responsible for providing pressure for blood flow and transporting blood to various parts of the body. The heart is composed of myocardial tissue, consisting of four chambers: the left atrium, left ventricle, right atrium, and right ventricle. The left and right atria and ventricles are separated by a septum, so they are not connected to each other. There are valves (atrioventricular valves) between the atria and ventricles, which allow blood to flow only from the atria into the ventricles and not backwards. The function of the heart is to promote blood flow, provide sufficient blood flow to organs and tissues, supply oxygen and various nutrients, and carry away metabolic end products (such as carbon dioxide, inorganic salts, urea, and uric acid), enabling cells to maintain normal metabolism and function. Heart microvascular endothelial cells are single-layer flat epithelioid cells that make up the surface of the heart microvascular lumen. The bioactive substances they produce and secrete play an important role in maintaining vascular tension, regulating blood pressure, and preventing thrombosis. They have significant pathophysiological significance in the pathogenesis of cardiovascular diseases. In recent years, a large number of studies have shown that the function and pathological changes of myocardial microvascular endothelial cells directly affect the function of myocardial cells, and they are also important targets for many toxins, inflammatory factors, viruses, and other factors. As an in vitro cell model for research, they have been used in myocardial ischemia-It plays an important role in the pathogenesis of reperfusion and the study of paracrine growth factors between cells.

Method Introduction:

Laboratory isolated porcine heart microvascular endothelial cellsCD31Immunofluorescence identification, purity can reach90%Above, and not containingHIV-1TheHBVTheHCVMycoplasma, bacteria, yeast, fungi, etc.

Quality inspection:

Laboratory isolated pig heart microvascular endothelium using collagenase-The mixed digestion method is combined with density gradient centrifugation, and then screened through endothelial cell specific culture medium. The total number of cells is approximately5×105cells/Bottle.

Training Information:

Package conditions:PLL（0.1mg/ml）Gelatin（0.1%）

Culture medium: Basic culture medium, includingFBSTheEGFThebFGFTheIGFTheVEGFTheHeparinTheHydrocortisoneThePenicillinTheStreptomycinwait

Fluid change frequency: every2-3Change the fluid once every day

Growth characteristics: Wall adhesion

Cell morphology: endothelial cell like

Passage characteristics: passable2-3generation

Digestive fluid:0.25%

Cultivation conditions: Gas phase: Air,95%；CO2，5%

Primary cells generally pass through about 10 generations, and most cells age and die. However, a very small number of cells can survive the "crisis" and continue to pass on, while surviving cells can generally pass through 40-50 generations, called cell lines. After 50 generations, there will be a "crisis" again, in which the genetic material of some cells has changed and has the characteristics of carcinogenesis, and may continue to be passaged indefinitely, called a cell line.

Some also believe that a cell line refers to a population of cells that have been successfully passaged from primary cell cultures. It also refers to cultured cells that can be continuously passaged for a long time. This led to the later development of finite cell lines and infinite cell lines. Therefore, cell lines refer narrowly to cells that can be continuously passaged, and broadly to cells that can be passaged. A cell line is a single cell obtained from primary culture or cell line through selection or cloning methods, which has special properties or markers, and is called a cell line.

Cell lines have advantages such as easy cultivation, wide variety, low cost, and unlimited passage. However, they are also prone to misidentification and cross contamination during cultivation and cryopreservation.

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The specific steps of primary cell culture:

1. Preparation: Take various disinfected culture products and place them on the purification table. Disinfect with ultraviolet light for 20 minutes. Wash your hands and wipe your elbows with 75% alcohol before starting work.

2. Layout: Ignite the alcohol lamp and install the straw cap.

3. Processing organization: Place the tissue block in a beaker and rinse 2-3 times with Hanks solution to remove blood stains; If you suspect that the organization may be contaminated, you can first place it in a mixture containing green for 30-60 minutes.

5. Digestion: Either use a constant temperature water bath or place in a 37 ℃ incubator for digestion. Shake every 20 minutes during digestion, and it is better to use an electromagnetic constant temperature stirrer for digestion. The digestion time depends on the size of the tissue block and the hardness of the tissue.

6. Separation: When the digestive fluid becomes turbid during the digestion process, a small amount of digestive fluid can be suctioned out with a pipette and observed under a microscope. If the tissue has dispersed into cell clusters or individual cells, digestion should be immediately terminated, and then the tissue blocks that have not been fully digested should be filtered out through a suitable stainless steel sieve. Centrifuge the digestion solution at low speed (500-1000 rpm) for 5 minutes, aspirate the supernatant, and add an appropriate amount of culture medium containing serum.

7. Counting: Use a counting plate to count. If the cell density in the cell suspension is too high, adjust it by adding culture medium and then divide it into culture bottles. For most cells, the pH requirement is in the range of 7.2-7.4, and the culture medium appears slightly red. If the color is yellowish, it indicates that the liquid has become acidic and can be adjusted with NaHCO3.

8. Cultivation: Incubate in a 37 ℃ incubator. If CO2 incubator is used for cultivation, the bottle mouth should be blocked with a gauze cotton stopper or a screw cap. The gauze stopper is prone to mold growth and needs to be replaced with a new stopper every time the liquid is changed.