Primary human brain vascular pericytes

Manufactured by ScienCell

Sourced in United States

Primary human brain vascular pericytes are cells that reside within the blood vessel walls of the brain. They play a core role in the regulation of blood flow and the maintenance of the blood-brain barrier.

Automatically generated - may contain errors

Lab products found in correlation

Penicillin, by Thermo Fisher Scientific (4 mentions) Streptomycin, by Thermo Fisher Scientific (4 mentions) Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (4 mentions) Hek293t cell, by American Type Culture Collection (2 mentions) Penicillin, by ScienCell (2 mentions) Streptomycin, by ScienCell (2 mentions) Pericyte specific growth medium, by ScienCell (2 mentions) Fetal bovine serum (fbs), by ScienCell (2 mentions) Recombinant human tgf β, by Thermo Fisher Scientific (1 mentions) Goat anti mouse cd31, by R&D Systems (1 mentions) Human astrocyte, by ScienCell (1 mentions) Ripa buffer, by Merck Group (1 mentions) Protease and phosphatase inhibitor cocktail, by Roche (1 mentions) Pericyte medium, by ScienCell (1 mentions) Poly l lysine (pll), by Bio-Techne (1 mentions) Laminin 521, by BioLamina (1 mentions)

7 protocols using primary human brain vascular pericytes

Culturing Primary Human Brain Pericytes

Check if the same lab product or an alternative is used in the 5 most similar protocols

Primary human brain vascular pericytes (ScienCell, Carlsbad, CA, USA, Cat# 1200) belonging to five different lots were maintained in pericyte-specific growth medium (ScienCell, Cat# 1201) supplemented with fetal bovine serum (FBS; 2%), growth factors, 100 units/mL penicillin, and 100 μg/mL streptomycin. Cells were used from passages 2 to 7. Human embryonic kidney (HEK)-293T cells (ATCC, Manassas, VA, USA, Cat# CRL-11268) were cultured in DMEM (Thermo Fisher Scientific, Carlsbad, CA, USA, Cat#11995–065) supplemented with 10% FBS (ScienCell, Cat# 0500), penicillin 100 units/mL, and 100 μg/mL streptomycin (Thermo Fisher Scientific, Cat# 15140–122). Cultures were maintained in 5% CO₂ at 37°C.

Torices S., Roberts S.A., Park M., Malhotra A, & Toborek M. (2020). Occludin, caveolin‐1, and Alix form a multi‐protein complex and regulate HIV‐1 infection of brain pericytes. The FASEB Journal, 34(12), 16319-16332.

+ Open protocol

+ Expand

Immunofluorescent Characterization of Pericytes

Check if the same lab product or an alternative is used in the 5 most similar protocols

Primary human brain vascular pericytes were obtained from ScienCell (Carlsbad, CA) and grown according to the vendor's instructions. Human recombinant TGF-β was purchased from Peprotech (100-21). Antibodies used in the immunofluorescent staining were purchased from commercial sources as follows: rabbit-anti-mouse collagen IV (Millipore, AB756P), mouse-anti-mouse-α-SMA-FITC (Sigma, F3777), goat-anti-mouse CD31 (R&D, AF3628), Alexa Fluor 555 conjugated goat-anti-Rabbit Ig (Invitrogen, A21429) and Alexa Fluor 488 conjugated goat-anti-human Ig (Invitrogen, A11013). Two monoclonal anti-CD248 antibodies, Clone 8 and 9G5, were generated in house by immunization of rabbits or rats (respectively) with a CD248ECD-Fc fusion protein. These antibodies were selected for all immunostaining and internalization assays as they do not compete with MORAb-004 for CD248 binding in a competition FACS assay (Figure S1C).

Rybinski K., Imtiyaz H.Z., Mittica B., Drozdowski B., Fulmer J., Furuuchi K., Fernando S., Henry M., Chao Q., Kline B., Albone E., Wustner J., Lin J., Nicolaides N.C., Grasso L, & Zhou Y. (2015). Targeting endosialin/CD248 through antibody-mediated internalization results in impaired pericyte maturation and dysfunctional tumor microvasculature. Oncotarget, 6(28), 25429-25440.

+ Open protocol

+ Expand

Vascular Pericytes and Astrocyte Cultures

Check if the same lab product or an alternative is used in the 5 most similar protocols

Primary human brain vascular pericytes and human astrocytes were obtained from ScienCell Research Laboratories (Carlsbad, CA, USA) and cultured for additional three passages before used for experiment. The cells were cultured in complete medium provided by the supplier, containing 2% fetal bovine serum (FBS), antibiotics (penicillin and streptomycin) and pericytes or astrocytes growth supplement. Primary central memory T (T_CM) cells were generated from naïve CD4⁺ T cells isolated from peripheral blood mononuclear cells (PBMC) selected from healthy blood donors, as previously described (Bosque and Planelles, 2009 (link); Piekna-Przybylska et al, 2017 (link)). ATM Kinase Inhibitor (KU55933) and PARP inhibitor I (3-ABA, 3-Aminobenzamide) were obtained from Calbiochem. DNA-PK inhibitor (NU7441) was obtained from Apexbio Technology LLC.

Piekna-Przybylska D., Nagumotu K., Reid D.M, & Maggirwar S.B. (2018). HIV-1 infection renders brain vascular pericytes susceptible to the extracellular glutamate. Journal of neurovirology, 25(1), 114-126.

+ Open protocol

+ Expand

Culturing Primary Human Brain Vascular Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols

Primary human brain vascular pericytes were purchased from ScienCell (Carlsbad, CA, USA) and cultured in the pericyte medium (provided by ScienCell). Cell culture dishes were coated with poly-L-lysine (2μg/cm²) and were used in passages 2–5. Human brain microvascular endothelial cells (HBMECs) were obtained also from ScienCell and cultured in endothelial medium (provided by ScienCell) and were used for passages 4–14.

Bai Y., Zhu X., Chao J., Zhang Y., Qian C., Li P., Liu D., Han B., Zhao L., Zhang J., Buch S., Teng G., Hu G, & Yao H. (2015). Pericytes Contribute to the Disruption of the Cerebral Endothelial Barrier via Increasing VEGF Expression: Implications for Stroke. PLoS ONE, 10(4), e0124362.

+ Open protocol

+ Expand

Culturing Primary Human Brain Pericytes

Check if the same lab product or an alternative is used in the 5 most similar protocols

Primary human brain vascular pericytes (ScienCell, Carlsbad, CA, USA, Cat# 1200) belonging to five different lots were maintained in pericyte‐specific growth medium (ScienCell, Cat# 1201) supplemented with fetal bovine serum (FBS; 2%), growth factors, 100 units/mL of penicillin, and 100 µg/mL of streptomycin. Cells were used from passages 2 to 7. Human embryonic kidney (HEK)‐293T cells (ATCC, Manassas, VA, USA, Cat# CRL‐11268) were cultured in DMEM (Thermo Fisher Scientific, Carlsbad, CA, USA, Cat#11995‐065) supplemented with 10% of FBS (ScienCell, Cat# 0500), penicillin 100 units/mL, and 100 µg/mL of streptomycin (Thermo Fisher Scientific, Cat# 15140‐122). Cultures were maintained in 5% of CO₂ at 37°C.

+ Open protocol

+ Expand

Isolation and Culture of Astrocytes and Pericytes

Check if the same lab product or an alternative is used in the 5 most similar protocols

Astrocytes were isolated and cultured from postnatal mouse brains (0–4 days) as described previously [61 (link)]. They were then seeded onto 24-well plates pre-coated with 0.001 % poly-l-lysine for 30 min. Primary human brain vascular pericytes (ScienCell) were also seeded in a similar way. Upon confluency, the cells were treated with 0.1 % BSA/PBS for 16 h which served as control, or with 500 ng/ml recombinant human Ang-2 for 16 h plus/minus 1 µM AKB-9785 for 10 min followed by washing with PBS. Cells were lysed for 15 min on ice in RIPA buffer (Sigma) containing one tablet each of protease and phosphatase inhibitor cocktail (Roche). Following this, the samples were transferred into Eppendorf tubes and centrifuged at 16,000×g at 4 °C for 10 min. The supernatants were transferred into new Eppendorf tubes and stored at −80 °C until further use.

Gurnik S., Devraj K., Macas J., Yamaji M., Starke J., Scholz A., Sommer K., Di Tacchio M., Vutukuri R., Beck H., Mittelbronn M., Foerch C., Pfeilschifter W., Liebner S., Peters K.G., Plate K.H, & Reiss Y. (2016). Angiopoietin-2-induced blood–brain barrier compromise and increased stroke size are rescued by VE-PTP-dependent restoration of Tie2 signaling. Acta Neuropathologica, 131, 753-773.

+ Open protocol

+ Expand

Engineered Human iPSC-Derived BBB Model

Check if the same lab product or an alternative is used in the 5 most similar protocols

For coculturing, astrocytes (0.12 × 10⁶ cells/cm², iCell Astrocyte; FCDI) and neurons (0.04 × 10⁶ cells/cm², iCell GABANeuron; FCDI) were cocultured on 24-well plates coated with 0.1 mg/mL poly-l-lysine (TREVIGEN, MD) and 3.3 μg/mL laminin-521 (BioLamina, Sweden) in 1:1 astrocyte/neuron medium purchased from FCDI on D4. On D8, 5.0 × 10³ cells/cm² of primary human brain vascular pericytes (ScienCell, CA) were seeded on the basolateral side of Transwells by flipping the plate upside-down in a pericyte medium (ScienCell). A coculture BBB model was constructed as shown in Figure 1B. In our human iPSC-derived BBB (hiBBB) models, ECM or neuron medium was used in each monoculture BBB system (Mono-ECM and Mono-N) or coculture BBB system (Co-ECM and Co-N), respectively. In the apical wells, ECM was used for all groups (Fig. 1A, B).

Ohshima M., Kamei S., Fushimi H., Mima S., Yamada T, & Yamamoto T. (2019). Prediction of Drug Permeability Using In Vitro Blood–Brain Barrier Models with Human Induced Pluripotent Stem Cell-Derived Brain Microvascular Endothelial Cells. BioResearch Open Access, 8(1), 200-209.

+ Open protocol

+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!