The I. scapularis ISE6 cell line was cultured in L15C300 medium supplemented with 10% heat-inactivated fetal bovine serum (FBS, 24 MilliporeSigma), 10% tryptose phosphate broth (BD), and 0.1% bovine lipoprotein concentrate (MP Biomedicals) at 34 °C [22 (link)]. Tick cells were grown to confluence and sub-cultured in capped T25 flasks (Greiner bio-one). All cell cultures were verified by PCR to be Mycoplasma-free (Southern Biotech).
Mycoplasma free
Manufactured by Southern Biotech
Mycoplasma-free is a laboratory equipment designed to detect and eliminate mycoplasma contamination in cell cultures. It provides a reliable and efficient solution for maintaining the integrity of cell-based research and production processes.
Automatically generated - may contain errors
Lab products found in correlation
Capped t25 flasks, by Greiner (2 mentions)
Glutamax, by Thermo Fisher Scientific (2 mentions)
Fetal bovine serum (fbs), by Merck Group (2 mentions)
Fetal bovine serum (fbs), by Gemini Bio (2 mentions)
6 well plate, by Merck Group (2 mentions)
24 well plate, by Corning (2 mentions)
L glutamine, by Quality Biological (2 mentions)
Bovine lipoprotein concentrate, by MP Biomedicals (1 mentions)
T25 flask, by Greiner (1 mentions)
Tryptose phosphate broth, by BD (1 mentions)
Rpmi 1640 medium, by Quality Biological (1 mentions)
Hl 60, by American Type Culture Collection (1 mentions)
3 protocols using mycoplasma free
1
Cultivation of Ixodes scapularis ISE6 Cells
2
Cultivation of Tick Cell Lines
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The I. scapularis IDE12 and ISE6 cell lines were obtained from Dr. Ulrike Munderloh at University of Minnesota. Cells were cultured in L15C300 medium supplemented with 10% heat inactivated fetal bovine serum (FBS, Sigma), 10% tryptose phosphate broth (TPB, Difco), 0.1% bovine cholesterol lipoprotein concentrate (LPPC, MP Biomedicals) at 34 °C in a non-CO2 incubator. ISE6 cells were grown to confluency in capped T25 flasks (Greiner bio-one) and either seeded at 3 × 106 cells/well in 6-well plates (Millipore Sigma) or 3 × 105 cells/well in 24-well plates (Corning). IDE12 cells were also grown to confluency in T25 flasks and either seeded at 1 × 106 cells/well in 6-well plates (Millipore Sigma) or 3 × 105 cells/well in 24-well plates (Corning). The human leukemia cell line, HL-60, was obtained from ATCC and maintained in RPMI-1640 medium with L-Glutamine (Quality Biological) supplemented with 10% FBS (Gemini Bio-Products) and 1% GlutaMax (Gibco), in vented T25 flasks (CytoOne) at 37 °C and 5% CO2. All cell cultures were confirmed to be Mycoplasma-free (Southern Biotech).
3
Culturing I. scapularis Cell Lines
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The I. scapularis IDE12 and ISE6 cell lines were obtained from Dr. Ulrike Munderloh at University of Minnesota. Cells were cultured in L15C300 medium supplemented with 10% heat inactivated fetal bovine serum (FBS, Sigma), 10% tryptose phosphate broth (TPB, Difco), 0.1% bovine cholesterol lipoprotein concentrate (LPPC, MP Biomedicals) at 34°C in a non-CO2 incubator. ISE6 cells were grown to confluency in capped T25 flasks (Greiner bio-one) and either seeded at 3×106 cells/well in 6-well plates (Millipore Sigma) or 3×105 cells/well in 24-well plates (Corning). IDE12 cells were also grown to confluency in T25 flasks and either seeded at 1×106 cells/well in 6-well plates (Millipore Sigma) or 3×105 cells/well in 24-well plates (Corning). The human leukemia cell line, HL-60, was obtained from ATCC and maintained in RPMI-1640 medium with L-Glutamine (Quality Biological) supplemented with 10% FBS (Gemini Bio-Products) and 1% GlutaMax (Gibco), in vented T25 flasks (CytoOne) at 37°C and 5% CO2. All cell cultures were confirmed to be Mycoplasma-free (Southern Biotech).
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