Pv11 cells, originally isolated from egg masses of P. vanderplanki, were cultivated in accordance with a previously published protocol [12 (link)]. Briefly, we cultivated the cells in IPL-41 medium (Gibco, Grand Island, NY, USA) supplemented with 10% fetal bovine serum (Hyclone, Logan, UT, USA) and 2.6 g/L tryptose phosphate broth. We obtained insect Sf9 cells derived from S. frugiperda, the fall armyworm (Lepidoptera), from Merck, and cultivated them in Sf900 medium (Gibco, Grand Island, NY, USA) without supplements. We maintained both Pv11 and Sf9 cultures in non-humidified incubators at 25 and 28 °C, respectively.
Ipl 41 medium
Manufactured by Thermo Fisher Scientific
Sourced in United States
IPL-41 medium is a culture medium designed for insect cell lines. It provides the necessary nutrients and components to support the growth and maintenance of insect cell cultures.
Automatically generated - may contain errors
Lab products found in correlation
Streptomycin, by Merck Group (7 mentions)
Penicillin, by Merck Group (7 mentions)
Amphotericin b, by Merck Group (7 mentions)
Tryptose phosphate broth, by BD (3 mentions)
Fetal bovine serum (fbs), by MP Biomedicals (3 mentions)
Tryptone phosphate broth, by BD (3 mentions)
Ipl 41 medium, by Merck Group (2 mentions)
Sf 900 medium, by Thermo Fisher Scientific (1 mentions)
X tremegene hp dna transfection reagent, by Roche (1 mentions)
Ham s f12 medium, by Fujifilm (1 mentions)
Penicillin streptomycin mixture, by Merck Group (1 mentions)
Antibiotic antimycotic solution, by Merck Group (1 mentions)
Bacto tryptose phosphate broth, by BD (1 mentions)
Fetal bovine serum (fbs), by Merck Group (1 mentions)
10 protocols using ipl 41 medium
1
Cultivation of Pv11 and Sf9 Insect Cells
2
Efficient RNA Interference in Silkworm Cells
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In this study, we used the cultured silkworm ovary-derived BmN4-SID1 cell line, which has been widely used for efficient RNA interference experiments in silkworms [49 (link)]. The BmN4-SID1 cell line was maintained at 27 °C in IPL-41 medium (Gibco, Waltham, MA, USA) supplemented with 10% fetal bovine serum (Hyclone, Logan, UT, USA). The expression plasmids for EGFP-CENP-N and FLAG-CENP-N were inserted into the genome of BmN4-SID1 cells using the piggyBac transposition system according to the previous report [48 (link)], and the stably transformed cells were selected by resistance to puromycin (CalBiochem, Darmstadt, Germany). For transient transfections, various plasmids were transfected into cells using X-tremeGENE HP DNA transfection reagent (Roche, Basel, Switzerland) according to the manufacturer’s instructions.
3
Cultivation of Pv11 Cell Line
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The Pv11 cell line isolated from the egg masses of P. vanderplanki was cultivated following previously published protocols45 (link). Briefly, we cultivated Pv11 cells in IPL-41 medium (Gibco, USA), supplemented with 10% foetal bovine serum (HyClone, USA) and 2.6 g/l tryptose phosphate broth, in a non-humidified incubator at 25 °C.
4
Culturing Pv11 and CHO Cells
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Pv11 cells and CHO cells were grown as previously described (9 (link), 14 (link), 82 (link)). Pv11 cells were cultured in IPL-41 medium (Thermo Fisher Scientific, Waltham, MA) supplemented with 2.6 g/L tryptose phosphate broth (Becton, Dickinson and Company, Franklin Lakes, NJ), 10% (v/v) fetal bovine serum (FBS) and 0.05% (v/v) of Antibiotic-Antimycotic Solution (10,000 units penicillin, 10 mg streptomycin and 25 μg amphotericin B per mL; Sigma–Aldrich, St. Louis, MO), designated hereafter as complete IPL-41 medium. A CHO-K1 derivative cell line, Flip-InTM-CHO cells (purchased from Thermo Fisher Scientific), was cultured in Ham’s F-12 medium (FUJIFILM Wako, Osaka, Japan) containing 10% (v/v) FBS, 100 units/mL penicillin and 100 μg/mL streptomycin (penicillin-streptomycin mixture; Sigma–Aldrich) at 37 °C, 5% CO2, and 95% relative humidity.
5
Pv11 Cell Line Culture Protocol
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Pv11 cells were originally established in our laboratory [14 (link)]. Pv11 cells and all clonal cell lines were grown in IPL-41 medium (Thermo Fisher Scientific, Waltham, MA, USA) supplemented with 2.6 g/L tryptone phosphate broth (Becton, Dickinson and Company, Franklin Lakes, NJ), 10% (v/v) fetal bovine serum (MP Biomedicals, Santa Ana, CA, USA), and 0.05% (v/v) of an antibiotic and antimycotic mixture (penicillin, amphotericin B, and streptomycin; Merck KGaA, Darmstadt, Germany).
6
Rearing P. vanderplanki Larvae for Research
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P. vanderplanki larvae were reared on 1% agar containing 2% commercial milk under controlled conditions (13 h light: 11 h dark, 27ºC) according to the previously described protocol (4 (link)). The strain used in this study was inbred for at least four generations (inbreeding code 4aG21b) and was registered as P. vanderplanki NIAS01. Larvae used in the experiments were starved for 24 h prior to DNA or RNA extraction. Pv11 cells were cultured in IPL-41 medium (Thermo Fisher Scientific, Waltham, MA) supplemented with 2.6 g/L Bacto™ Tryptose Phosphate Broth (Becton, Dickinson and Company, Franklin Lakes, NJ), 10% (v/v) fetal bovine serum (US origin; MP Biomedicals, Santa Ana, CA) and 0.05% (v/v) of an antibiotic and antimycotic mixture (penicillin, amphotericin B and streptomycin; Merck KGaA, Darmstadt, Germany). Cell passage was conducted every seven days.
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P. vanderplanki larvae were reared on 1% agar containing 2% commercial milk under controlled conditions (13 h light: 11 h dark, 27ºC) according to the previously described protocol (4 (link)). The strain used in this study was inbred for at least four generations (inbreeding code 4aG21b) and was registered as P. vanderplanki NIAS01. Larvae used in the experiments were starved for 24 h prior to DNA or RNA extraction. Pv11 cells were cultured in IPL-41 medium (Thermo Fisher Scientific, Waltham, MA) supplemented with 2.6 g/L Bacto™ Tryptose Phosphate Broth (Becton, Dickinson and Company, Franklin Lakes, NJ), 10% (v/v) fetal bovine serum (US origin; MP Biomedicals, Santa Ana, CA) and 0.05% (v/v) of an antibiotic and antimycotic mixture (penicillin, amphotericin B and streptomycin; Merck KGaA, Darmstadt, Germany). Cell passage was conducted every seven days.
7
Pv11 and Pv11-KH Cell Culture Protocol
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Pv11 and Pv11-KH cells were grown as described previously [18 (link)]. Briefly, for their culture, IPL-41 medium (Thermo Fisher Scientific, Waltham, MA, USA) supplemented with 2.6 g/L tryptose phosphate broth (Becton, Dickinson and Company, Franklin Lakes, NJ, USA), 10% (v/v) fetal bovine serum, and 0.05% (v/v) of an antibiotic and antimycotic mixture (penicillin, amphotericin B, and streptomycin; MilliporeSigma, Burlington, MA, USA) was used, and the medium is designated hereafter as complete IPL-41 medium. A density of 3 × 105 cells/mL were seeded into a cell culture flask with plug seal cap and grown at 25 °C for 6–7 days.
8
Culturing and Trehalose Treatment of Pv11 Cells
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Pv11 cells were originally established in our laboratory [27 (link)]. The cell culture was maintained as described [15 (link)]. Briefly, Pv11 cells were cultured using IPL-41 medium (Thermo Fisher Scientific, Waltham, MA, USA) supplemented with 2.6 g/L tryptone phosphate broth (Becton, Dickinson and Company, Franklin Lakes, NJ, USA), 10% (v/v) fetal bovine serum (MP Biomedicals, Santa Ana, CA, USA) and 0.05% (v/v) of an antibiotic and antimycotic mixture (penicillin, amphotericin B, and streptomycin; Merck KGaA, Darmstadt, Germany). As trehalose treatment, the cells were incubated in trehalose mixture (600 mM trehalose (Hayashibara Co., Ltd., Okayama, Japan) containing 10% (v/v) IPL-41 medium).
9
Culturing Pv11 and Pv11-KH Cell Lines
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Pv11 and Pv11-KH cells were grown in IPL-41 medium (Thermo Fisher Scientific, Waltham, MA) supplemented with 2.6 g/L tryptose phosphate broth (Becton, Dickinson and Company, Franklin Lakes, NJ), 10% (v/v) fetal bovine serum, and 0.05% (v/v) of an antibiotic and antimycotic mixture (penicillin, amphotericin B, and streptomycin; MilliporeSigma, Burlington, MA)5 (link), designated hereafter as complete IPL-41 medium.
10
Pv11 Cell Culture and Trehalose Treatment
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Pv11 cells were originally established in our laboratory [27] . The cell culture was maintained as described [15] (link). Briefly, Pv11 cells were cultured using IPL-41 medium (Thermo Fisher Scientific, Waltham, MA) supplemented with 2.6 g/L tryptone phosphate broth (Becton, Dickinson and Company, Franklin Lakes, NJ), 10% (v/v) fetal bovine serum (MP Biomedicals, Santa Ana, CA), and 0.05% (v/v) of an antibiotic and antimycotic mixture (penicillin, amphotericin B, and streptomycin; Merck KGaA, Darmstadt, Germany). As trehalose treatment, the cells were incubated in trehalose mixture (600 mM trehalose (Hayashibara Co., Ltd., Okayama, Japan) containing 10% (v/v) IPL-41 medium).
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