Primary cells of human heart microvascular endothelial cells

Primary cells of human heart microvascular endothelial cells

Training Information:

Package conditionsPLL（0.1mg/ml）Gelatin（0.1%）

culture mediumcontainFBSGrowth additivesPenicillinTheStreptomycinwait

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

Growth characteristics: Wall adhesion

Cell morphology endothelial cell like

Passage characteristics can be passed down2-3generation

Digestive fluid0.25%

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

Isolation of microvascular endothelial cells from human 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, with oxygen and various nutrients, and carry away metabolic end products (such as carbon dioxide, inorganic salts, urea, and uric acid), allowing 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:

The human heart microvascular endothelial cells isolated in the company laboratory were treated with collagenase-Prepared by combining protease digestion with density gradient centrifugation, and then screened through endothelial cell specific culture medium, the total number of cells is approximately5×10?cells/Bottle.

Quality inspection:

The human heart microvascular endothelial cells isolated from the company laboratoryCD31Immunofluorescence identification, purity can reach90%Above, and not containingHIV-1TheHBVTheHCVMycoplasma, bacteria, yeast, fungi, etc.

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, arrangement, and mutual influence between cells, as well as the biological regulatory effects of local environments.

CM-R034Rat hepatic artery endothelial cells, Chinese hamster ovary cellsK1(Subtype cloning);CHO-K1 SMPD1 Others MousemouseSMPD1 / ASMRod-shaped virus-Insect cell lysate PLAT Others MousemousetPA / PLATHuman cell lysate mouse primary glomerular endothelial cells rabbit primary glomerular endothelial cells

beta-Amyloid(1-16 0.5mgbeta-Amyloid(1-16) peptide (rat, mouse)β - amyloid peptide(1-16)(Rats, mice)

ITGA8 & ITGB1 Protein Humanrecombinant humanITGA8 & ITGB1 Heterodimerprotein

HPXRecombinant miceHemopexin / HPXproteinProtein

IL12B Protein RatRecombinant ratsIL12B / IL-12Bprotein

NRG1recombinant humanNRG1-alphaprotein(ECD, Fclabel) Protein

Human heterogeneous ribonucleoprotein complex/resistRA33antibody(HNP / RA 33) ELISAtest kit

Human ansfer factor (TF) ELISA KitHuman transfer factor(TF)test kit

Humahymidylatesyhetase,TSELISAKitHuman TS(TS)test kit96T/48TImport packaging

HumanIerleukin17,IL-17Reagent kit for human interleukin17(IL-17)Kit specifications:96T/48T

Organic soluble total protein preparation kit20time

Humaniestinalfattyacidbindingprotein，iFABPELISAKitHuman intestinal fatty acid binding protein(iFABP)Kit specifications:96T/48T

Human progesterone/pregnant(PROG)ELISAKit ELISA. 96T/48T

Human hypersensitive free estriol(UE 3) elists ELISA. 96T/48T

Human norepinephrine(that) elisay ELISA. 96T/48T

Human prostaglandinF(PGF)ELISAKit ELISA. 96T/48T

Primary cells of human heart microvascular endothelial cellsGuinea pig interleukin12(IL-12/P70)ELISAtest kit96T/48T

Human cytochrome P450c21B/21- hydroxylase (CYP21B) ELISA KitHuman cytochromeP450c21B/21-hydroxylase(CYP21B)test kit

Humanai-Bactericidalpermeabilityincreasingproteinaibody,BPI-AbELISAKitHuman antimicrobial permeability protein antibody(BPI-Ab)test kit96T/48TImport packaging

HumanCyclosporineA,CsAReagent kit personnelA(CsA)Kit specifications:96T/48T

organizationHCV(HEPATITISVIRUSC)Virus qualitative test kit20time

Humanpokeweedmitogen，% s's neighbourhoodHuman PWM(PWM)Kit specifications:96T/48T

HAVCR1Recombinant crab eating monkeyKIM-1 / TIM1 / HACVR1proteinProtein

MTR-1A (Melatonin receptor-1A 0.5mgMTR-1A (Melatonin receptor-1A)Melatonin receptor(antigen)

PDE4Brecombinant humanPDE4B / DPDE4protein(His & GSTlabel) Protein

CFL1 Protein Humanrecombinant humanCFL1 / N-cofilinprotein

ENTPD5 Protein MouseRecombinant miceEntpd 5protein

After receiving the cells, please proceed with the following steps for the customer. (The number of passages for this cell is limited to about 2. It is recommended that the customer arrange subsequent experiments as soon as they receive the cells.)

1. Remove the cell vial, disinfect it with 75% alcohol, remove the sealing film, and place it in a 37 ℃, 5% CO2 incubator for 6-8 hours to stabilize the cell state.

2. When the cells reach 80% confluence, prepare for subculture.

3. Cell passage:

1) Suck out the culture medium from the cell vial and wash the cells once with PBS.

2) Add about 1mL of 0.125% digestive solution to the culture bottle and digest at 37 ℃ for about 3 minutes; Under the microscope, after the cells shrink and become round, discard the digestion solution and add culture medium to terminate digestion.

3) Gently blow and mix with a straw, inoculate and passage in appropriate ratios such as 1:2 or 1:3, then add fresh culture medium to 5mL and culture in a 37 ℃ 5% CO2 cell incubator.

4) After the cells adhere to the wall, culture and observe, and replace with fresh culture medium every 2-3 days.