Human placental trophoblastic cell lines (JEG-3 and HTR-8/SVneo) were purchased from Procell (Wuhan, China) and KALANG (Shanghai, China), respectively. Cells were cultured in DMEM (Biosun, Shanghai, China) plus 10% fetal bovine serum at 37°C in an atmosphere of 5% CO 2 .
Htr 8 svneo
Manufactured by Procell
Sourced in China
The HTR-8/SVneo is a laboratory equipment designed for high-temperature heat treatment and sintering applications. It features a programmable temperature control system and a compact, durable design.
Automatically generated - may contain errors
Lab products found in correlation
Jeg 3, by Procell (2 mentions)
Lipofectamine 2000, by Thermo Fisher Scientific (2 mentions)
Htr 8 svneo, by Thermo Fisher Scientific (2 mentions)
Dmem h medium, by Thermo Fisher Scientific (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (1 mentions)
Htr8 svneo, by Abcam (1 mentions)
Lipofectamine 2000 reagent, by Thermo Fisher Scientific (1 mentions)
Rpmi 1640 medium, by Thermo Fisher Scientific (1 mentions)
Fetal bovine serum (fbs), by Merck Group (1 mentions)
Penicillin streptomycin, by Merck Group (1 mentions)
Oe nc, by GenePharma (1 mentions)
Tunicamycin, by Abcam (1 mentions)
Mir nc, by Sangon (1 mentions)
Anti mir nc, by Sangon (1 mentions)
Si nc, by Sangon (1 mentions)
Insulin, by Yuanye Bio-Technology (1 mentions)
Anisomycin, by Yuanye Bio-Technology (1 mentions)
Fetal bovine serum (fbs), by Sartorius (1 mentions)
Insulin, by Solarbio (1 mentions)
9 protocols using htr 8 svneo
1
Cultivation of Placental Trophoblast Cells
2
Hypoxia Regulation of Trophoblast Cells
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The trophoblast cell line HTR8/SVneo were purchased from Procell Life Science & Technology Co., Ltd. (Wuhan, China.) and cultured in DMEM-H medium (Gibco, USA) containing, 100 U/ml Penicillin–Streptomycin (Gibco, USA), and 10% fetal bovine serum (FBS, Gibco, USA). Cells which cultured at 37℃ and 5%CO2 conditions were adopted as normal control. And cells in the hypoxia group were placed under hypoxia (1% O2) in anoxic box, which can freely adjust oxygen concentration and flow meter detection, closed and cultured at 37℃ and 5%CO2. Cells were collected 48 h later for subsequent experiments.
3
Gestational Diabetes Trophoblast Modeling
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The human trophoblast cell line HTR8/SVneo was obtained from Procell Life Science & Technology Co., Ltd. HTR8/SVneo cells were grown in RPMI-1640 medium (Gibco; Thermo Fisher Scientific, Inc.) supplemented with 10% fetal bovine serum (MilliporeSigma) and 1% penicillin-streptomycin (MilliporeSigma) in a humidified incubator at 37°C in an atmosphere of 5% CO2. The HTR8/SVneo cells were treated with 25 mM HG for 24 h to mimic the in vitro gestational diabetic environment and treated with 78 ng/ml tunicamycin (Abcam) for 16 h. HTR8/SVneo cells maintained in media containing 5 mM glucose were used as the control group.
pcDNA3.1(+) CTRP9 overexpression vector (Oe-CTRP9) and empty vector NC (Oe-NC; Shanghai GenePharma, Co., Ltd.) at a concentration of 20 µM were transfected into HTR-8/SVneo cells using Lipofectamine® 2000 reagent (Invitrogen; Thermo Fisher Scientific, Inc.), in accordance with the manufacturer's instructions. Cells were transfected at 37°C for 8 h and were used in subsequent experiments 48 h post-transfection.
pcDNA3.1(+) CTRP9 overexpression vector (Oe-CTRP9) and empty vector NC (Oe-NC; Shanghai GenePharma, Co., Ltd.) at a concentration of 20 µM were transfected into HTR-8/SVneo cells using Lipofectamine® 2000 reagent (Invitrogen; Thermo Fisher Scientific, Inc.), in accordance with the manufacturer's instructions. Cells were transfected at 37°C for 8 h and were used in subsequent experiments 48 h post-transfection.
4
Modulating FAM99A and YAP1 in Trophoblasts
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Human placenta trophoblast cell line HTR8/SVneo was bought from Procell (China) and cultivated in RPMI-1640 medium (PM150110A; Procell) with 5% fetal bovine serum (FBS; Procell) in 5% CO2 in an incubator at 37°C.
Small interfering RNA targeting FAM99A (si-FAM99A#1, si-FAM99A#2, and si-FAM99A#3), FAM99A overexpression plasmid (FAM99A), YAP1 overexpression plasmid (YAP1), miR-134-5p mimics (miR-134-5p), miR-134-5p inhibitor (anti-miR-134-5p), and their corresponding negative controls (si-NC, miR-NC, and anti-miR-NC) were all obtained from Sangon Biotech (China). Cell transfection was conducted using Lipofectamine 2000 (Invitrogen, USA).
Small interfering RNA targeting FAM99A (si-FAM99A#1, si-FAM99A#2, and si-FAM99A#3), FAM99A overexpression plasmid (FAM99A), YAP1 overexpression plasmid (YAP1), miR-134-5p mimics (miR-134-5p), miR-134-5p inhibitor (anti-miR-134-5p), and their corresponding negative controls (si-NC, miR-NC, and anti-miR-NC) were all obtained from Sangon Biotech (China). Cell transfection was conducted using Lipofectamine 2000 (Invitrogen, USA).
5
Human Trophoblast Cell Cultivation
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The human extravillous trophoblast cell line HTR-8/SVneo and the choriocarcinoma cell lines JAR and JEG-3 were purchased from Procell Life Science & Technology Co., Ltd. All cells were maintained in Ham's F 12 medium (Hyclone) supplemented with 10% heat-inactivated FBS and 1% penicillin-streptomycin at 37°C in a 5% CO2 incubator.
6
Insulin Resistance Induction and JNK Signaling Regulation
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Immortalized human chorionic trophoblast line HTR-8/SVneo was purchased from Procell (Wuhan, Hubei, China), and cultured in RPMI-1640 medium supplemented with 5% fetal bovine serum (FBS) (Biological industries, Kibbutz Beit-Haemek, Israel) at 37°C with 5% CO2.
The transfection was performed using Lipofectamine 2000 (Invitrogen, Carlsbad, CA, USA) with serum-free medium.
The cells were incubated with insulin (Solarbio) of 10-6 mol/l for 48 h to induce IR, and insulin of 10-7 mol/l was used to stimulate cells to verify the IR.
JNK signaling agonist Anisomycin (1 μg/ml) (Yuanye, Shanghai, China) and antagonist SP600125 (20 μM) (Aladdin, Shanghai, China) was used to treat cells for 30 min to activate or inactivate the JNK signaling.
The transfection was performed using Lipofectamine 2000 (Invitrogen, Carlsbad, CA, USA) with serum-free medium.
The cells were incubated with insulin (Solarbio) of 10-6 mol/l for 48 h to induce IR, and insulin of 10-7 mol/l was used to stimulate cells to verify the IR.
JNK signaling agonist Anisomycin (1 μg/ml) (Yuanye, Shanghai, China) and antagonist SP600125 (20 μM) (Aladdin, Shanghai, China) was used to treat cells for 30 min to activate or inactivate the JNK signaling.
7
Culturing Human Placental Trophoblast Cells
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Human placenta trophoblast cells (HTR-8/SVneo) were purchased from Procell (Wuhan, China) and maintained in Roswell Park Memorial Institute-1640 (RPMI-1640; Biosun, Shanghai, China) added with 10% fetal bovine serum (FBS; Biosun) and 1% penicillin/streptomycin (Phygene, Fuzhou, China) at 37°C in a humid incubator with 5% CO2.
8
Visualizing FITC-ROPPIP Uptake in HTR-8/Svneo Cells
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Human trophoblast cells (HTR-8/Svneo) were acquired from Wuhan Procell Life Technology Co., Ltd. (catalog number: CL-0599, Wuhan, China). The cells were cultured with Roswell Park Memorial Institute (RPMI) 1640 media (Sigma, USA) supplemented with 10% fetal bovine serum (FBS), 100 U/mL penicillin, and 100 mg/mL streptomycin double-antibody, which was placed in a constant temperature and humidity incubator containing 5% carbon dioxide (CO2) at 37 °C. The culture media was changed every day and the cells were passaged when they reached 90% confluence. HTR-8/Svneo cells were administrated with 100 µM FITC-ROPPIP and incubated for 1 h at 37 °C in the dark, then imaged with a Confocal laser scanning microscopy (magnification, ×200).
9
Gestational Diabetes Mellitus Cell Model
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Human trophoblast cells (HTR-8/SVneo) provided by Procell (Wuhan, China) were cultured in RPMI1640 medium containing 10% FBS at 37°C incubator mixed with 5% CO 2 . Cell model of GDM was constructed by treating HTR-8/SVneo cells with HG (30 mM; Sigma-Aldrich, St. Louis, MO, USA) for 24 h as previously described [16] [17] [18] . Cells in the control group were treated with normal glucose (5.5 mM).
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