Primary cells of human dermal microvascular endothelial cells

Primary cells of human dermal 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 juice0.25%

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

Human dermal microvascular endothelial cells isolated from dermal tissue; The dermis is located deep in the epidermis and is connected to subcutaneous tissue downwards. The collagen fibers and elastic fibers of the dermal connective tissue are interwoven and buried in the matrix. It is distributed with various connective tissue cells and a large number of collagen fibers and elastic fibers, making the skin both elastic and resilient. It consists of two layers: the nipple layer and the mesh layer. The structural composition of genuine leather is collagen, elastic fibers, and matrix. Microvascular endothelial cells（MEC）It plays a very important role in the inflammatory response, tumor growth, and wound healing process. In the field of wound healing research, due to the maturity of in vitro culture techniques for epidermal cells and dermal fibroblasts, extensive and in-depth research has been conducted on these two types of cells. In contrast, research on the other main cell involved in wound healing, dermal microvascular endothelial cells, is limited due to the difficulty of their in vitro isolation and culture techniques.

Method Introduction:

The human dermal 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:

Human dermal 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 focus on observing the connections, arrangements, and interactions between cells, as well as the biological regulatory effects of local environments.

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.

COC1Human ovarian cancer cells SCaBER(Human bladder squamous cell carcinoma cells) RNASET2 Others HumanpersonRNASET2Rod-shaped virus-Insect cell lysate SHH Others HumanpersonSHH / Sonic hedgehog (aa 1-197)Mouse primary renal proximal tubular epithelial cells derived from human cell lysate and rat primary olfactory bulb neuronal cells

Placental alkalinity0acid enzyme(ALPP)recombinant proteinRecombinant Alkaline Phosphatase, Placental (ALPP)

HIST1H3A Protein HumanRecombinant humanHistone H3.1 / HIST1H3A / H3FAprotein

KLK13Recombinant humanKLK13 / Kallikrein-13proteinProtein

HA Protein H11N2Recombinant influenza AH11N2 (A/duck/Yangzhou/906/2002)hemagglutinin(Hemagglutinin / HA)protein

SEMA3ARecombinant humanSemaphorin 3A / SEMA3Aprotein(Fclabel) Protein

Rat tissue metalloproteinase inhibitor4(TIMP4)Reagent kit, English name:TIMP4 ELISA Kit

Mouse ai thyroid stimulating hormone receptor aibody (Ab) ELISA KitMouse anti thyroid stimulating hormone receptor antibody(Ab)test kit

MousehepatitisBviruscoreaibody,HBcAbELISAKitMouse hepatitis B core antibody(HBcAb)test kit96T/48TImport packaging

CLIAKitforHumanMaixGlaProtein,MGPELISAKitHuman matrix gamma carboxyglobulin

transmission electron microscope(TEM)Production processing embedding fluidALiquid:10milliliterBLiquid:30milliliter

ELISAKitGzms-BRat granule enzymeB

Rabbit element(Renin)ELISAKit ELISA. 96T/48T

Rabbit insulin-like growth factor2 (igf- 2) elists ELISA. 96T/48T

Rabbit insulin-like growth factor1 (igf- 1) elists ELISA. 96T/48T

Rabbit insulin(INS)ELISAKit ELISA. 96T/48T

Primary cells of human dermal microvascular endothelial cellsMouse hyaluronic acid(HA)ELISAtest kit96T/48T

Human heparan sulfate (HS) ELISA KitHuman heparin(HS)test kit

Humanformylmethionine,fMetELISAKitHuman formyl sulfide(fMet)test kit96T/48TImport packaging

ChickensolubleE-selectin,sE-selectinChicken soluble reagent kitEchoice of plain food(sE-selectin)Kit specifications:96T/48T

Glass slide cellsPDGF-BProtein expression fluorescence microscopy kit10/20time

Mouseferritin,FEELISAKitMouse ferritin(FE)Kit specifications:96T/48T

HARecombinant influenza AH7N9 (A/Shanghai/1/2013)hemagglutininHA1 (Hemagglutinin)proteinProtein

phospho-eIF4E(pSer209 0.5mgphospho-eIF4E(pSer209)(phospho-Eukaryotic translation initiation factor 4E) 0acidificationeIF-4E(Ser209)antigen

PVRL1Recombinant humanCD111 / Nectin-1 / PVRL1proteinProtein

HEXB Protein HumanRecombinant humanHexosaminidase B / HEXBprotein

CTSA Protein HumanRecombinant humanCathepsin A / CTSAprotein