All four sphere-cultured cells were implanted in 6–8-week-old NOD/SCID mice (Charles River Laboratories, Wilmington, MA). Briefly, 3×103 patient-derived sphere cancer cells were suspended in 100ul Matrigel (BD Biosciences, San Jose, CA) and injected into the subcutaneous layer on the backs of NOD/SCID mice. After 3–4 months, when the tumor size reached >1cm3, the mice were anesthetized via an intraperitoneal injection of a mixture of 40mg/kg Zoletil (Virbac Laboratories, BP 27-06511 Carros, France) and 5mg/kg Rum pun (Bayer Korea, South Korea), and the tumors were surgically removed.
100ul matrigel
Manufactured by BD
Sourced in United States
Matrigel is a solubilized basement membrane preparation extracted from the Engelbreth-Holm-Swarm (EHS) mouse sarcoma, a tumor rich in extracellular matrix proteins. The product is supplied as a 100 microliter volume.
Automatically generated - may contain errors
2 protocols using 100ul matrigel
1
Patient-Derived Xenograft Tumor Establishment
2
Matrigel-Coated Invasion Assay
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Invasion assay was performed using invasion chambers with 6.4 mm diameters and 8 mm pore size (Corning, USA) coated with 100 ul matrigel (BD, USA). Briefly, cells resuspended in DMEM culture medium with no FBS were seeded into the upper chamber. DMEM medium containing 20% FBS was filled into the lower chamber as the chemoattractant. After 48 h of incubation, the non-invasive cells were removed from the upper side of the chamber, and the invasive cells were stained using crystal violet. The chamber was then observed and photographed under a light microscope to count invasive cells.
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