Murine Cancer Cell Lines and Macrophages

A murine colon cancer cell line Colon26 (aka CT26) [23 (link),67 (link)] and a murine calvaria-derived osteoblastic cell line MC3T3-E1 [68 (link)] were obtained from Cell Bank at RIKEN BioResource Research Center (Tsukuba, Japan). A rapidly metastatic subline, LuM1, was generated from Colon26 as described previously [23 (link),35 (link),36 (link),42 (link),43 (link)]. MMP3-knockout LuM1 (MMP3-KO) cells were generated using the genome-editing method as described below. A murine macrophage-like cell line RAW-D (a subline of RAW264.7) was kindly provided by Prof. Toshio Kukita (Kyushu University, Fukuoka, Japan) [69 (link)]. Colon26, LuM1, MMP3-KO LuM1, and MC3T3-E1 were maintained in RPMI1640 with 10% fetal bovine serum (FBS) and penicillin, streptomycin, and amphotericin B. RAW-D cells were cultured in minimum essential medium (MEM; Wako Pure Chemicals, Osaka, Japan) containing 10% FBS, supplemented with penicillin (100 U/mL) and streptomycin (100 mg/mL).

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Okusha Y., Eguchi T., Tran M.T., Sogawa C., Yoshida K., Itagaki M., Taha E.A., Ono K., Aoyama E., Okamura H., Kozaki K.I., Calderwood S.K., Takigawa M, & Okamoto K. (2020). Extracellular Vesicles Enriched with Moonlighting Metalloproteinase Are Highly Transmissive, Pro-Tumorigenic, and Trans-Activates Cellular Communication Network Factor (CCN2/CTGF): CRISPR against Cancer. Cancers, 12(4), 881.

Publication 2020

MC3T3-E1 MEM

Amphotericin b Calvaria Cell line Cells Colon cancer Cultured medium D cells Fetal bovine serum Macrophage Mmp3 Murine Penicillin Raw264 7 cell line Streptomycin

Corresponding Organization : Okayama University

Other organizations : Harvard University, Beth Israel Deaconess Medical Center, Tokushima University, Ain Shams University, Okayama University Hospital

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Variable analysis

independent variables

Generation of MMP3-knockout LuM1 (MMP3-KO) cells using the genome-editing method

dependent variables

Not explicitly mentioned

control variables

Colon26 cell line
LuM1 cell line
MC3T3-E1 cell line
RAW-D cell line
Cell culture conditions (RPMI1640 with 10% FBS and penicillin, streptomycin, and amphotericin B for Colon26, LuM1, MMP3-KO LuM1, and MC3T3-E1; MEM with 10% FBS, penicillin, and streptomycin for RAW-D)

positive controls

Not specified

negative controls

Not specified

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