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2.1. Cell Culture and Establishment of MTX-Resistant Osteosarcoma Subclones

JZ Jingyong Zhang
SW Shubao Wang
YB Yang Bai
AA Aasi Mohammad Ali
JD Jiewen Deng
YC Yushi Chen
YF Yonghui Fu
MH Ming He
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The human osteoblast hFOB1.19 (catalog: CRL-11372), human osteosarcoma cell lines U2OS (catalog: HTB-96), and MG63 (catalog: CRL-1427) were sourced from the American Type Culture Collection (Manassas, VA, USA). U2OS and MG63 were cultured in Dulbecco’s modified Eagle medium (catalog: BE12-604F, Lonza, Basel, Switzerland) supplemented with 10% fetal bovine serum (catalog: F7524, Sigma-Aldrich, St. Louis, MO, USA). hFOB1.19 cells were cultivated in hFOB1.19 complete medium (catalog: EP-ML-0353, Elabscience, Wuhan, China). All cells were kept at 37 °C in a 5% CO2 environment.

Parental U2OS and MG63 cells were initially exposed to 3 ng/mL (0.01 µM) MTX [15]. Subjecting the parental cell lines to stepwise raised MTX concentrations, MTX-resistant subclones were produced.

Copyright and License information:  ©2023 by the authors.
Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).

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