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5 protocols using jimt 1 cells

1

JIMT-1 Cell Culture Protocol

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JIMT-1 cells obtained from AddexBio (San Diego, CA, USA) were cultured in Dulbecco’s modified Eagle’s medium (DMEM) (Corning, Corning, NY, USA) supplemented with 10% heat-inactivated fetal bovine serum (Gibco, Waltham, MA, USA), 1% L-glutamine, and 1% penicillin/streptomycin (Corning, Corning, NY, USA). Cells were maintained in a 37 °C humidified incubator with 5% CO2.
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2

Comparative Growth of Breast Cancer Cell Lines

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JIMT-1 cells were purchased from Addexbio (Cat #C0006005, San Diego, CA, USA) propagated and maintained in DMEM-F12 (ATCC Cat# 11320-033, Manassas, VA, USA) with fetal bovine serum added to a final concentration of 10%. BT-474 cells were purchased from ATCC (Cat # HTB-20, Manassas, VA, USA) propagated and maintained in RPMI 1640 media (ATCC Cat# 30-2001, Manassas, VA, USA) with fetal bovine serum added to a final concentration of 10%. Both cell lines (Figure 1) were grown in tissue culture flasks in a humidified atmosphere of 95% air and 5% CO2 at 37 °C. Growth medium was renewed every 2 or 3 days and cell densities were maintained at 1 × 105 cells/mL before passage. Under these incubation conditions it had been determined that after seeding of equal amounts of cells, the rates of cell division of the JIMT-1 cells were ~ 2X that of the BT-474 cells.
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3

Optically Driven Cell Repositioning

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Example 4

Medium:

Serum free medium (ThermoFisher Scientific, Cat. No. 12045-096), with a conditioning culture medium additive, B-27® Supplement (2% v/v).

FIGS. 12A-C depict experimental results from before and after optically driven displacement and repositioning of JIMT-1 cells (commercially available from AddexBio Cat. # C0006055), an adherent human breast carcinoma cell line. FIG. 12A showed a series of sequestration pens of microfluidic device 1200 prior to the displacement of cells 1204 from sequestration pen 1202. Optical illumination was directed towards sacrificial feature 1220 (thermal target), a surface of the sequestration pen 1202, and cells 1204 were dislodged. FIG. 14B showed an individual cell 1204a, dislodged and newly repositioned in an empty sequestration pen adjacent to originally occupied sequestration pen 1202. FIG. 14C shows a timepoint 20 h after displacement and repositioning showing that cells 1204a were viable and have doubled from one cell to two, within the 20 h culture period.

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4

Cytotoxic Effects of Doxorubicin and Dexrazoxane

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Doxorubicin HCl (DOX) was purchased from Selleck Chemicals (Houston, TX) while Dexrazoxane (DEX) was from Millipore Sigma-Aldrich Co. (St. Louis, MO). DOX was dissolved in molecular biology grade water while DEX in DMSO as per manufacturer’s instructions. Stock solutions were stored at −80°C for long-term storage while fresh serial dilutions were prepared in cell culture media each time prior to experiments. JIMT-1 cells were procured from AddexBio (San Diego, CA) and cultured in cell culture media comprising of Dulbecco’s Modified Eagle’s Medium (DMEM) with 10% sterile filtered fetal bovine serum (FBS), 1% sodium bicarbonate, 1% MEM Non-essential amino acids, and 1% penicillin/streptomycin antibiotics. MDA-MB-468 cells were purchased from American Type Culture Collection (ATCC) (Manassas, VA) and cultured in DMEM with 10% FBS, and 1% penicillin/streptomycin. Both the cell lines were maintained at 37°C in a humidified atmosphere with 5% CO2 and passaged upon confluency with 0.25%Trypsin/2.21 nM EDTA. Details regarding the remaining reagents have been previously described elsewhere (Mody et al., 2023 (link)).
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5

JIMT-1 Cell Culture Protocol

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JIMT-1 cells were acquired from the AddexBio (San Diego, CA, United States) and maintained in Dulbecco’s Modified Eagle’s Medium (DMEM; Corning Inc., Tewksbury, MA, United States) media supplemented with 10% fetal bovine serum (FBS; Sigma–Aldrich, St. Louis, MO, United States), 2 mM glutamine, sodium pyruvate (Corning Inc.), sodium bicarbonate (Corning Inc.), and penicillin-streptomycin solution (Corning Inc.). Cells were cultured at 37°C, 5% CO2, and split to maintain confluency.
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