Human tendon derived cells were harvested from patellar tendon during tendon grafting operations after obtaining written informed consent. From these tissue specimens human primary tenocytes were isolated and cultured. All cells were maintained in Dulbecco's Modified Eagle's Medium (DMEM/F-12 with Glutamax, Gibco-BRL) supplemented with penicillin (100 U ml−1), streptomycine (10 μg ml−1) (both Sigma-Aldrich) and 10% fetal calf serum (Gibco-BRL). Cells were detached by incubation with 0.05% trypsin for 5 min at 37°C. They were identified as tenocytes through their characteristic growth pattern and by detection of scleraxis (SCX) and tenomodulin (TNMD) expression. The studies were performed with cells at passage 2–3 and a total of 3 donors were used for all assays.
Streptomycine
Manufactured by Merck Group
Sourced in Germany, United States
Streptomycine is a laboratory equipment product manufactured by Merck Group. It is a general-purpose tool used for various applications in scientific research and analysis. The core function of Streptomycine is to facilitate the detection and identification of streptococcus bacteria in clinical and research samples.
Automatically generated - may contain errors
Lab products found in correlation
Penicillin, by Merck Group (4 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (2 mentions)
Fetal calf serum (fcs), by Thermo Fisher Scientific (1 mentions)
Dmem f12 with glutamax, by Thermo Fisher Scientific (1 mentions)
Leukemia inhibitory factor, by Merck Group (1 mentions)
Trypsin, by Thermo Fisher Scientific (1 mentions)
Neomycin, by Merck Group (1 mentions)
Tyrode s acid, by Merck Group (1 mentions)
Genticin, by Merck Group (1 mentions)
Pmsg folligon, by MSD (1 mentions)
High pure pcr purification kit, by Roche (1 mentions)
Mitomycin c, by Merck Group (1 mentions)
Pmsg folligon, by MSD animal health (1 mentions)
Falcon 3047, by BD (1 mentions)
Glutamax 1, by Thermo Fisher Scientific (1 mentions)
Ez pcr mycoplasma test kit, by Sartorius (1 mentions)
Lncap 1f5, by Merck Group (1 mentions)
Lncap, by Merck Group (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Rpmi 1640 medium, by Merck Group (1 mentions)
4 protocols using streptomycine
1
Isolation and Characterization of Human Tenocytes
2
Generation of Knockout Mice via ES Cells
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Mitomycin C, Genticin, Penicillin, Streptomycine, Neomycin, M16 and M2 media, Plasmid purification kit, Tyrodes acid and leukemia Inhibitory Factor were purchased from Sigma (Germany). PMSG (Folligon) and hCG (Chorulon) were purchased from MSD Animal Health (New Zealand). DMEM, FCS and trypsin was obtained from Gibco BRL (France). RNA and DNA extraction kits were bought from Intron (Korea). Endofree Plasmid Maxi Kit was purchased from Qiagene (USA). High Pure PCR Purification Kit was from Roche (Germany). EW and R1 embryonic stem cells and pEGFP-C1 were kindly donated by Dr. Karim Nayernia at Institute of Molecular Medicine and Cell Therapy, Düsseldorf, Germany and GENEOCELL, Canada.
To generate knockout mouse, ES cells were cultured and electroporated with the gene targeting vector. The genetically-modified cells were selected and transferred into embryos. The modified ES cells along with the cells of embryo were used to produce chimeric mice. Crosses between the chimeras and wild type mice were resulted heterozygous offspring.
To generate knockout mouse, ES cells were cultured and electroporated with the gene targeting vector. The genetically-modified cells were selected and transferred into embryos. The modified ES cells along with the cells of embryo were used to produce chimeric mice. Crosses between the chimeras and wild type mice were resulted heterozygous offspring.
3
Rat Pancreatic Acinar Tumor Cell Line Culture
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The study was performed on Rat Pancreatic Acinar Tumor Cell Line—AR42J cells (American Type Culture Collection, Rockville, MD, USA). Cell culture was grown in the RPMI 1640 Medium supplemented with Glutamax-I (Gibco BRL, Gaithersburg, MD, USA) and 10% fetal bovine serum (FBS, heat-inactivated; Gibco-BRL, Grand Island, NY, USA) with addition of 100 U/mL penicillin and 100 µg/mL streptomycine (Sigma-Aldrich, St. Louis, MO, USA) in the standard conditions: 37 °C and 5% CO2 [127 (link),128 (link),129 (link),130 (link)].
Twenty-four hours before the experiments, culture medium was replenished with fresh RPMI 1640 containing 2% FBS and without antibiotics. Cells cultures were plated at the initial density of 2 × 106/mL in a 100 mm culture plate (Falcon 3047; Becton Dickinson, Lincoln Park, NJ, USA) and allowed to attach for 12 h. AR42J cells were incubated in GHRL-free or GHRL containing medium and in the presence or absence of caerulein or in a combination of the above. GHRL was given at a concentration of 10−7 M, caerulein at a concentration of 10−8 M, incubation time was 48 h. All experiments were repeated at least six times.
Twenty-four hours before the experiments, culture medium was replenished with fresh RPMI 1640 containing 2% FBS and without antibiotics. Cells cultures were plated at the initial density of 2 × 106/mL in a 100 mm culture plate (Falcon 3047; Becton Dickinson, Lincoln Park, NJ, USA) and allowed to attach for 12 h. AR42J cells were incubated in GHRL-free or GHRL containing medium and in the presence or absence of caerulein or in a combination of the above. GHRL was given at a concentration of 10−7 M, caerulein at a concentration of 10−8 M, incubation time was 48 h. All experiments were repeated at least six times.
4
Cell Line Cultivation and Authentication
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The LNCaP, 22Rv1, PC3, VCaP, RWPE1, RWPE2 and 293 T cell lines were originally obtained from the American Type Culture Collection (ATCC), and have been authenticated by STR profiling. The LNCaP-1F5 and V16A cell lines were kindly provided by Prof. Hansen He, University of Toronto and Prof. Olli A Jänne, University of Helsinki, respectively. All cell lines were tested regularly for mycoplasma (EZ-PCR Mycoplasma Test Kit, 20–700-20, Biological Industries). All cell lines were found to be negative of mycoplasma during our study. LNCaP, LNCaP-1F5, V16A, PC3 and 22Rv1 cells were cultured in RPMI1640 medium (Sigma), VCaP and 293T cells were grown in DMEM (Invitrogen), RWPE1 and RWPE2 cells were cultured in Keratinocyte-Serum Free Medium (KSF, Invitrogen). All mediums were supplemented with 10% FBS (Gibco) and 1% antibiotics (penicillin and streptomycine, Sigma), with the exception of KSF medium that was supplemented with the epidermal growth factor (EGF) and Bovine Pituitary Extract (BPE) included in the kit (17,005–042, Introvigen). To induce androgen signaling and AR activity, relevant PCa cells were treated with dihydrotestosterone (DHT, Sigma) for indicated time.
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