Conventional culture dishes that are 50 mm in diameter were purchased from Falcon (Sigma-Aldrich, Milan, Italy). Leibovitz’s L-15 medium, α-MEM GlutaMAX medium, insulin transferrin selenium (ITS) 100×, and LIVE/DEAD Fixable Far Red stain were purchased from Invitrogen (Milan, Italy). Penicillin-streptomycin 100×, amphotericin B 250 μg/mL, bovine serum albumin, L-ascorbic acid, L-glutamine 200 mM, Hoechst 33342, fructose, and α-thioglycerol were purchased from Sigma-Aldrich (Milan, Italy). Mayers’s hematoxylin and paraffin wax were purchased from Carlo Erba (Milan, Italy).
Insulin transferrin selenium 100
Manufactured by Thermo Fisher Scientific
Sourced in Italy
Insulin transferrin selenium (ITS) 100× is a concentrated supplement for cell culture media. It provides insulin, transferrin, and selenium, which are important components for cell growth and proliferation.
Automatically generated - may contain errors
Lab products found in correlation
Leibovitz s l 15 medium, by Thermo Fisher Scientific (2 mentions)
Bovine serum albumin (bsa), by Merck Group (2 mentions)
Fructose, by Merck Group (2 mentions)
Hoechst 33342, by Merck Group (2 mentions)
α thioglycerol, by Merck Group (2 mentions)
L ascorbic acid, by Merck Group (2 mentions)
Amphotericin b 250 μg ml, by Merck Group (2 mentions)
L glutamine 200 mm, by Merck Group (2 mentions)
Live dead fixable far red stain, by Thermo Fisher Scientific (1 mentions)
Penicillin streptomycin 100, by Merck Group (1 mentions)
α mem glutamax medium, by Thermo Fisher Scientific (1 mentions)
Paraffin wax, by Carlo Erba (1 mentions)
Eosin y, by Merck Group (1 mentions)
Ldh kit, by Merck Group (1 mentions)
Dulbecco s modified eagle medium, by Thermo Fisher Scientific (1 mentions)
Gentamycin, by Merck Group (1 mentions)
Recombinant human egf, by Thermo Fisher Scientific (1 mentions)
Newborn calf serum nbcs, by Thermo Fisher Scientific (1 mentions)
Penicillin, by Lonza (1 mentions)
Streptomycin, by Lonza (1 mentions)
4 protocols using insulin transferrin selenium 100
1
Cell culture protocol for viability
2
Optimized Tissue Culturing Protocols
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Lumox culture dishes with gas-permeable bottom 50 mm in diameter (PD) were purchased from Sarstedt AG & Co. KG (Nuembrecht, Germany). Conventional dishes 50 mm in diameter were purchased from Falcon (Sigma-Aldrich, Milan, Italy). Leibovitz's L-15 medium, α-MEM Glutamax medium (code number 32571), insulin transferrin selenium (ITS) 100×, and live/dead fixable Far Red stain were purchased from Invitrogen (Milan, Italy). Penicillin streptomycin (Pen-Strep) 100×, amphotericin B 250 μg/ml, bovine serum albumin BSA, L-ascorbic acid, L-glutamine 200 mM, Hoechst 33342, fructose, α-thioglycerol, and eosin-Y were purchased from Sigma Aldrich (Milan, Italy). Mayers's hematoxylin and paraffin wax were from Carlo Erba (Milan, Italy). Estradiol and progesterone solid phase enzyme immunoassay kits were obtained from DRG (Marburg, Germany). LDH kit was obtained from Merck (Sigma-Aldrich, Milan, Italy). The tissue chopper was purchased from McIlwain, Mickle Laboratory Engineering Company Ltd (Surrey, UK). Pump tubing was purchased from Cole-Parmer (Milan, Italy) and the peristaltic pump from Ismatec (Enco, Venice, Italy).
3
Isolation and Culture of Primary HCEC and HCSF
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Human donor eyes were received from the Georgia Eye Bank (Atlanta, GA, USA) within 24 hours after death. Primary HCEC and HCSF were isolated from the donors’ eyes according to the established protocols74 (link),75 (link). HCEC were cultured in high glucose DMEM (Dulbecco’s modified eagle medium, Gibco), 10 ng/mL human recombinant EGF (epidermal growth factor), 10% fetal bovine serum, 1% insulin-transferrin-selenium-100× (Gibco), and 40 µg/mL gentamycin (Sigma). The HCSF were cultured similarly as HCEC but without human recombinant EGF.
4
Synthesis and Characterization of Heterocyclic Compounds
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The imidazo[2,1-b][1,3,4]thiadiazole derivative IS1 and the indole derivatives IS2, IS3 and IS4 were synthesised at the Department of Pharmacy, University of Palermo, Italy, following the synthetic procedures previously described[39 (link),49 (link)]. The compounds were dissolved in dimethyl sulfoxide (DMSO), as reported previously[41 (link)]. Gemcitabine was kindly provided by Eli Lilly Corporation (Indianapolis, IN, USA) and dissolved in sterile water. Cell medium and newborn calf serum (NBCS) were from Gibco (Gaithersburg, MD, USA), while penicillin (50 IU mL-1) and streptomycin (50 µg mL-1) were from Lonza (Switzerland). Insulin-transferrin-selenium 100× was from Gibco (Grand Island, NY, USA) and PB was purchased from Cayman Chemical (Ann Arbor, MI, USA). All other chemicals were purchased from Sigma-Aldrich (Zwijndrecht, The Netherlands).
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