Thp 1

AML cell lines with t(8;21) chromosome translocation (Kasumi-1 and SKNO-1) were used. Kasumi-1 was obtained from American Type Culture Collection (ATCC, CRL-274) and cultured in RPMI1640 media (Gibco, Waltham, MA) supplemented with 20% fetal bovine serum (FBS). SKNO-1 was obtained from the JCRB cell bank (Osaka, Japan, JCRB 1170) and cultured in RPMI1640 supplemented with 10% FBS and 10ng/mL recombinant human granulocyte-macrophage colony-stimulating factor (GM-CSF) (Peprotech, Rocky Hill, NJ). THP-1 was obtained from the Korean Cell Line Bank (Seoul, South Korea, 40202) and cultured in RPMI 1640 media supplemented with 0.05mM 2-mercaptoethanol (Gibco) and 10% FBS.

The human monocyte cell line, THP-1, and CRC cell lines (HCT15 and Caco2) were obtained from the Korean Cell Line Bank. The cells were grown in Roswell Park Memorial Institute (RPMI)-1640 medium (HyClone, Logan, UT, USA) containing 10% fetal bovine serum (FBS), 1% penicillin (100 U/mL) and streptomycin (100 mg/mL; Gibco, Grand Island, NY, USA), at 37 °C, in a humid environment with 5% CO₂.

Unless indicated otherwise, all cell culture products and plastics were supplied by WELGENE (Gyeongsanbuk-do, Republic of Korea) and SPL LIFE SCIENCE (Gyeonggi-do, Republic of Korea). For BMDMs, bone marrow cells were isolated from the tibia and femur of mice (C57BL/6 mice, 6–12 weeks old, NARA BIOTECH, Seoul, Republic of Korea), and incubated in the DMEM media containing fetal bovine serum (FBS, 10%), antibiotics and L929 cell-conditioned media (30%, a source of a macrophage colony-stimulating factor) for 7 days. Human monocyte-like cells (THP-1; KOREA CELL LINE BANK, Seoul, Republic of Korea) were differentiated into macrophages in RPMI 1640 (RPMI) containing FBS (10%), antibiotics, and phorbol 12-myristate 13-acetate (200 nM, PMA; INVIVOGEN, San Diego, CA, USA) for 24 h. All cells were incubated at 37 °C in an atmosphere containing 5% CO₂.

SW480, U937, THP‐1, Jurkat T (clone E6‐1), and CT26 cells were purchase from Korean Cell Line Bank (KCLB, Seoul, Korea). SW480, U937, THP‐1, and Jurkat T cells were maintained in complete RPMI media with 10% FBS and CT26 was maintained in complete Dulbecco's Modified Eagle's Medium (DMEM) media with 10% FBS. MC38 cell was purchase from Kerafast (Boston, MA) and it was maintained in complete DMEM media with 10% FBS. ROS induced senescent tumor cells were generated by H₂O₂ (200 × 10⁻⁶m) treatment for 4 days.

The human lung epithelial cell line A549 (CCL-185™) was purchased from ATCC (Manassas, VA, USA), and the human monocyte cell line THP-1 (40202) was purchased from Korean Cell Line Bank (Seoul, South Korea). A549 cells were cultured in RPMI 1640 medium (Welgene Inc., Daegu, South Korea) supplemented with 10% fetal bovine serum (GE Healthcare Life Sciences, Victoria, Australia), 50 U/mL penicillin, and 50 μg/mL streptomycin (Life Technologies, Carlsbad, CA), at 37°C in an incubator with 5% CO₂. THP-1 cells were cultured in RPMI 1640 medium supplemented with 10% fetal bovine serum, as described above.

MCF7, Jukart (Korean Cell Line Bank), and differentiated THP-1 (THP-1, Korean Cell Line Bank) were exposed to either vehicle or Cas9-loaded EVs for 2 h at 37 °C. The cells were collected and fixed. A flow cytometer examined the expression of Cas9-GFP in the harvested cells.