Cell migration was evaluated using a Bio-Coat cell migration chambers (BD Biosciences, MA, USA), which consist of a 24-well companion plate with cell culture inserts containing a filter with 8 μm-diameter pores. Briefly, 2 × 10 4 cell resuspended in 500 μL of EGM-2 medium (Gibco, USA) were seeded in the upper chamber, then vascular endothelial growth factor (VEGF) in EGM-2 medium at 100 mg/L was placed in the lower compartment of the chamber. After incubation for 24 h, the upper surface of the membrane was wiped with a cotton-tipped applicator to remove non-migrating cells, the migrating cells on the lower surface were fixed with 2% paraformaldehyde and stained with Giemsa solution. Migrating cells were counted manually in three random microscopic fields.
Egm 2 medium
Manufactured by Thermo Fisher Scientific
Sourced in United States
The EGM-2 medium is a cell culture medium designed for the growth and maintenance of various mammalian cell lines. It provides the necessary nutrients and supplements to support cell proliferation and viability in in vitro cell culture applications.
Automatically generated - may contain errors
Lab products found in correlation
Fetal bovine serum (fbs), by Thermo Fisher Scientific (2 mentions)
Matrigel, by BD (2 mentions)
Pen strep, by Thermo Fisher Scientific (1 mentions)
α mem, by Thermo Fisher Scientific (1 mentions)
Protein lysis buffer, by Solarbio (1 mentions)
Lactoferrin, by Merck Group (1 mentions)
Elisa kit, by Nanjing Jiancheng (1 mentions)
Huvec cell line, by American Type Culture Collection (1 mentions)
Cell counting kit 8 cck 8, by Solarbio (1 mentions)
Matrigel, by Thermo Fisher Scientific (1 mentions)
U tv0.5xc 3, by Olympus (1 mentions)
Bio coat cell migration chamber, by BD (1 mentions)
5 protocols using egm 2 medium
1
Cell Migration Assay Protocol
2
Hypoxia-Induced Ischemic Injury in HUVECs
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HUVECs were cultured in glucose-free EGM-2 medium (Gibco, USA) for 8 h at 37 °C in a hypoxic incubator (1% O2, 5% CO2) to induce ischemic injury.
3
Rat Hepatocyte and HUVEC Co-Treatment
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Rat hepatocytes (WB-F344) were cultured in α-MEM (GIBCO BRL, Langley, OK, USA) medium, supplemented with 1% penicillin, streptomycin (Pen-Strep, GIBCO BRL), and 5% fetal bovine serum (FBS, GIBCO BRL). Human umbilical vein endothelial cells (HUVECs) were cultured in EGM-2 medium at (GIBCO BRL) 37℃, in a 5% CO2 incubator. Additionally, cells were treated with siRNA-CRP and scrambled siRNA for 12 hours followed by treatment with 100 µM lithocholic acid (LCA, Sigma Aldrich) for 12 hours.
4
Lactoferrin Modulates HUVEC Angiogenesis
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Lactoferrin, of 95% purity, was purchased from Sigma (St. Louis, MO, USA). The HUVEC cell line was purchased from the American Type Culture Collection (ATCC, Manassas, VA, USA). EGM-2 medium and fetal bovine serum (FBS) were obtained from Gibco (Waltham, MA, USA); 1% streptomycin/penicillin was purchased from Thermo Fisher (Waltham, MA, USA). Cell Counting Kit-8 (CCK8) was purchased from Solarbio (Beijing, China). ELISA kits were purchased from Jiancheng (Nanjing, China). The PDXP siRNA fragment was synthesized by Sangon (Shanghai, China). The primary and secondary antibodies were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Protein lysis buffer and the related reagents were purchased from Solarbio. Enhanced chemiluminescence (ECL) reagent was purchased from Tanon (Shanghai, China).
5
In vitro Yolk Sac Vessel Formation from Stem Cells
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To investigate the potential formation of yolk sac vessels from stem cells, an in vitro test was performed using the BD Matrigel™ substrate (BD Biosciences, USA). As a positive control, bovine umbilical vein endothelial cells (BUVECs) cultured in vitro under the same conditions as those of bYS-MSCs. The Matrigel (BD Biosciences) was prepared according to the manufacturer's instructions.
The bYS-MSCs were plated at a concentration of 1 × 105 cells in EGM-2 medium (Gibco-BRL, USA) with 10% FCS on the Matrigel substrate (Covas et al., 2008). Samples were viewed under phase-contrast microscopy (Olympus IX71), and pictures were recorded using a digital camera (Olympus U-TV0.5XC-3) at 0 hour, 10 hours, 24 hours, 5 days, 10 days, and 15 days. The area was quantified using the ImageJ software (NIH Image-BioLab). We did the comparative analysis of the umbilical cord and bYS-MSCs.
The bYS-MSCs were plated at a concentration of 1 × 105 cells in EGM-2 medium (Gibco-BRL, USA) with 10% FCS on the Matrigel substrate (Covas et al., 2008). Samples were viewed under phase-contrast microscopy (Olympus IX71), and pictures were recorded using a digital camera (Olympus U-TV0.5XC-3) at 0 hour, 10 hours, 24 hours, 5 days, 10 days, and 15 days. The area was quantified using the ImageJ software (NIH Image-BioLab). We did the comparative analysis of the umbilical cord and bYS-MSCs.
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