Tumour perivascular cells (TPC) were isolated from human colorectal cancer tissues that were collected after surgical excision at the First Affiliated Hospital of Jinan University (Supplementary Table 1 ). The tissues were washed with pre‐chilled Hank's balanced salt solution (HBSS, Gibco) to remove surface blood, adipose tissues, and tissue fragments. Tumour vessels were separated and minced in pre‐chilled HBSS before endothelial cells were removed under a dissecting microscope. The rinsed vessel pieces were digested with tumour dissociation kit (Miltenyi Biotec) and then performed with gentleMACS Dissociators (Miltenyi Biotec) according to the manufacturer's instructions. The dissociated cells were washed with phosphate buffer saline (PBS, HyClone) twice, followed by culture in Pericyte Medium (PM, ScienCell) containing 2% fetal bovine serum (FBS, ExCell Bio), 1% Pericyte Growth Supplement (PGS), and 1% penicillin‐streptomycin. The cells were maintained in the atmosphere at 37℃ with 5% CO2. Adherent cells were collected and identified with transmission electron microscope (TEM) analysis, immunofluorescence assay, and flow cytometry (FCM) analysis.
Pericyte medium pm
Manufactured by ScienCell
Sourced in United States
Pericyte Medium (PM) is a specialized culture medium formulated to support the growth and maintenance of pericytes in vitro. It is designed to provide the necessary nutrients and growth factors required for the optimal proliferation and differentiation of pericyte cells.
Automatically generated - may contain errors
Lab products found in correlation
Gentlemacs dissociator, by Miltenyi Biotec (1 mentions)
Hanks balanced salt solution (hbss), by Thermo Fisher Scientific (1 mentions)
Fetal bovine serum (fbs), by ExCell Bio (1 mentions)
Tumour dissociation kit, by Miltenyi Biotec (1 mentions)
Hyaluronidase, by Merck Group (1 mentions)
Collagenase, by Merck Group (1 mentions)
Malondialdehyde tetra butyl ammonium salt mda, by Merck Group (1 mentions)
Penicillin, by ScienCell (1 mentions)
Streptomycin, by ScienCell (1 mentions)
Xtt cell viability assay kit, by Merck Group (1 mentions)
Matrigel, by BD (1 mentions)
Hrp conjugated anti rabbit igg 7074, by Cell Signaling Technology (1 mentions)
Pkh26, by Merck Group (1 mentions)
Pkh67, by Merck Group (1 mentions)
Dual luciferase reporter gene assay kit, by Promega (1 mentions)
Mg132, by Targetmol (1 mentions)
Endothelial cell medium (ecm), by ScienCell (1 mentions)
Cycloheximide (chx), by Targetmol (1 mentions)
Alexa fluor 594 conjugated donkey anti rabbit igg ab150064, by Abcam (1 mentions)
Chip pcr kit, by Merck Group (1 mentions)
8 protocols using pericyte medium pm
1
Isolation and Characterization of Colorectal Cancer Tumour Perivascular Cells
2
Isolation and Culture of Pericyte-Derived Cells
3
Evaluating Pericyte Viability under Aβ and MDA
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The human brain vascular pericyte (HBVP) cell line were cultured at 37°C in 5% CO2, 95% air in Pericyte medium (PM) (ScienceCell Inc.) containing 4% FBS, 1% Pericyte growth supplement (PGS), 100 U/mL penicillin and 100 μg/mL streptomycin (ScienceCell Inc.). HBVP are characterized by using antibody specific to | |-smooth muscle actin and are negative for HIV-1, HBV, HCV, mycoplasma, bacteria, yeast, and fungi. In all experiments, cells were grown to 70-90% confluence and subjected to a maximum of 20 cell passages. For the metabolic activity assay, 1×104 HBVP were incubated with 0.5 μg of human soluble Aβ 1-40 monomers (cat. num.: AS-24236-5, AnaSpec) and/or 1 mM of malondialdehyde tetrabutylammonium salt (MDA; cat. num.: 63287, Sigma Aldrich) for 24 h prior to the absorbance based “XTT Cell viability” assay kit (cat. num.: 9095, Cell Signaling). The cell viability assays were performed accordingly to the manufacturer's protocol.
4
Angiogenesis Regulation in Endothelial Cells
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Endothelial cell medium (ECM) and pericyte medium (PM) were products of ScienCell Research Laboratories (San Diego, CA, USA). Antibodies against Ki67 (#12,202), α-SMA (#19,245), bFGF (#20,102), FGFR1 (#76,123), FGFR2 (#23,328), β-actin (#23,328) and HRP-conjugated anti-rabbit IgG (#7074) were obtained from Cell Signaling Technology (Danvers, MA, USA). Antibodies against Gli1 (AF3455) and CD31 (AF3628), and a Proteome Profiler Human Angiogenesis Array Kit (ARY007) were obtained from R&D Systems (Minneapolis, MN, USA). Alexa Fluor 594-conjugated donkey anti-rabbit IgG (ab150064) and Alexa Fluor488- conjugated donkey anti-mouse IgG (ab150073) were obtained from Abcam (Cambridge, UK). GANT-61, cycloheximide (CHX) and MG132 were purchased from TargetMol (Shanhai, China). Matrigel was obtained from BD Biosciences (Franklin Lakes, NJ). Recombinant human bFGF was purchased from BioVision (Palo Alto, CA, USA). A ChIP PCR kit was obtained from Merck Millipore (Billerica, MA, USA). A dual luciferase reporter gene assay kit was provided by Promega (Madison, WI, USA). Rhodamine-labeled lysinated dextran (70-kDa) was product from Thermo Fisher Scientific (Waltham, MA, USA). PKH 26, PKH 67 and other reagents were obtained from Sigma-Aldrich (St. Louis, MO, USA).
5
Cultivation of Human Brain Cell Types
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Human brain vascular pericytes (abbreviated as HBVP; obtained from ScienCell Research Laboratories, Carlsbad, CA, USA) were cultured on poly‐l ‐lysine‐coated dishes in Pericyte Medium (PM; ScienCell Research Laboratories) supplemented with 5% FBS (Sigma‐Aldrich, St. Louis, MO, USA) and penicillin/streptomycin solution (P/S; ScienCell Research Laboratories) and used between passage numbers 2 and 6. Human astrocytes (HA; ScienCell Research Laboratories) were grown on poly‐l ‐lysine‐coated dishes in astrocyte medium (AM) supplemented with 5% FBS and P/S (all from ScienCell Research Laboratories) and used between passage numbers 2 and 4. Human microvascular cerebral endothelial cells (hCMEC/D3, shortly D3) were cultured on rat‐tail collagen‐coated dishes in endothelial cell basal medium‐2 (EBM‐2) with EGM‐2MV kit including supplements and 2% FBS (all from Lonza, Basel, Switzerland) and used between passage numbers 30 and 40.
6
Isolation and Culturing of Arf-GFP Pericytes
7
Magnetic Nanocapsule Biocompatibility Evaluation
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Biological tests with the resulting magnetic nanocapsules were conducted in both the human cerebral microvascular endothelial hCMEC/D3 (Merck Millipore, SCC006) and Human Brain Vascular Pericytes HBVP (ScienCell, 1200) cell lines. hCMEC/D3 cells were cultured in EndoGRO-MV-VEGF Complete Culture Media Kit (Merck Millipore, SCME003) supplemented with 1% penicillin-streptomycin (P/S, Gibco). HBVP cells were cultured in Pericyte Medium (PM, ScienCell, 1201).
8
Isolation and Characterization of Stem Cells
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The SHEDs were purchased from ALLCELLS (Alameda, CA, USA), and the DPSCs were provided by Songtao Shi (Department of Anatomy and Cell Biology, University of Pennsylvania School of Dental Medicine), and their use was approved by the Ethics Committee of the University of Pennsylvania. Stem cell phenotypic markers, CD45, CD90, CD105, and CD73, were used to evaluate SHED and DPSC stemness using flow cytometry.
Human brain vascular pericytes (cat. #1200) and human umbilical vein endothelial cells (HUVECs) (Cat. #8000) were purchased from ScienCell (Carlsbad, CA, USA) and were cultured in Pericyte Medium (PM, ScienCell) and fully supplemented with endothelial growth medium (EGM-2, Lonza Walkersville, MD, USA), respectively.
Human brain vascular pericytes (cat. #1200) and human umbilical vein endothelial cells (HUVECs) (Cat. #8000) were purchased from ScienCell (Carlsbad, CA, USA) and were cultured in Pericyte Medium (PM, ScienCell) and fully supplemented with endothelial growth medium (EGM-2, Lonza Walkersville, MD, USA), respectively.
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