Telomerase-immortalized human endometrial stromal cells (T-HESC, ATCC CRL-4003) [31 (link)] were cultured in DMEM/Nutrient Mixture Ham's F-12 with l-glutamine (Sigma Aldrich, St. Louis, MO), 1% BD ITS+ Universal Culture Supplement Premix (BD Biosciences, San Jose, CA), 1.5 g/liter sodium bicarbonate, 2% penicillin / streptomycin, and 10% charcoal-treated FBS (HyClone), which is referred to as HESC medium herein. For the cell experiments, T-HESCs were decidualized (dT-HESCs) as described [19 (link), 20 (link)] by incubating the cells with 0.5 mM 8-bromo-cyclic amp (cAMP) (Sigma Aldrich) for three to six days. Completely confluent monolayers were used for all experiments to ensure that only decidual cell surfaces were available for bacterial interactions.
T hesc
Manufactured by Merck Group
Sourced in United States
T-HESCs are a type of laboratory equipment used for the culture and maintenance of induced pluripotent stem cells (iPSCs). They provide a controlled environment for the growth and differentiation of these specialized cells, which are used in various research and therapeutic applications.
Automatically generated - may contain errors
Lab products found in correlation
T hesc, by American Type Culture Collection (4 mentions)
Dmem ham s nutrient mixture f 12, by Merck Group (2 mentions)
8 bromo cyclic amp camp, by Merck Group (2 mentions)
L glutamine, by Merck Group (2 mentions)
Medroxyprogesterone acetate mpa, by Merck Group (2 mentions)
Its premix, by BD (1 mentions)
8 bromoadenosine 3 5 cyclic monophosphate 8 br camp, by Merck Group (1 mentions)
Stempro 34, by Thermo Fisher Scientific (1 mentions)
Carbenoxolone, by Merck Group (1 mentions)
Phenol red, by Sartorius (1 mentions)
Suramin, by Santa Cruz Biotechnology (1 mentions)
Foetal bovine serum (fbs), by Sartorius (1 mentions)
Apyrase, by Merck Group (1 mentions)
Penicillin streptomycin, by Sartorius (1 mentions)
Its g, by Thermo Fisher Scientific (1 mentions)
Arl67156, by Merck Group (1 mentions)
Adenosine triphosphate (atp), by Merck Group (1 mentions)
Its universal cell culture supplement premix, by BD (1 mentions)
Crl 4003, by American Type Culture Collection (1 mentions)
6 protocols using t hesc
1
Decidualization of Immortalized Endometrial Cells
2
In Vitro Decidualization of T-HESCs
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The T-HESCs were purchased from ATCC (CRL-4003TM). T-HESCs were cultured in DMEM/F12 medium without phenol red (D2906, Sigma) supplemented with 10% charcoal/dextran treated fetal bovine serum (SH30068.03, HyClone), 1.5 g/L sodium bicarbonate, 1% ITS+ Premix (354,352, BD), and 500 ng/mL puromycin. T-HESCs were grown in an incubator at 37 °C under a 5% CO2 atmosphere at constant humidity. To generate an in vitro decidualization model, T-HESCs were cultured in 2% charcoal/dextran treated FBS medium with or without 1 μM MPA and 0.5 mM 8-Br-cAMP [26 (link)]. After three days, the culture medium was renewed with the same treatment. On day 6, the decidualization was confirmed by observing the enlarged rounded cell shape under a microscope and measuring the expressions of IGFBP-1 and PRL by quantitative reverse-transcription PCR (qRT-PCR) or the IGFBP-1 protein in the supernatant by ELISA.
3
Optimized Culture and Decidualization of Trophoblast Cells
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LAD2 cells (kindly provided by Dr. Kirschbaum, passages 3 - 10) were cultured in StemPro-34 containing recombinant human stem cell factor (SCF; Peprotech, Rocky Hill, NJ, USA) (25 (link)). HTR-8/SVneo cells (kindly provided by Dr. Graham, passage 3 - 12) (26 (link)) and T HESCs (purchased from the American Type Culture Collection; Manassas, VA, USA, passages 3 - 10) were cultured in RPMI1640 medium supplemented with 10% fetal bovine serum, 2 mM L-glutamine, 100 units/mL penicillin, and 100 μg/mL streptomycin. Decidualization of T HESCs was induced by culturing with 1 µM medroxyprogesterone acetate (MPA; #M1629; Sigma-Aldrich, St. Louis, MO, USA) and 0.5 mM 8-bromoadenosine 3’:5’-cyclic monophosphate (8-Br cAMP; #B7880; Sigma) for 5–6 days (27 (link)). We confirmed the decidualization of T HESCs based on the presence of morphological changes and increased PRL mRNA.
4
Culturing and Treating Endometrial Cell Lines
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T‐HESCs (human telomerase reverse transcriptase‐immortalized human endometrial stromal cells) and AN3‐CA cells were purchased from American Type Culture Collection (ATCC). T‐HESCs were cultured in DMEM/F12 medium without phenol red (Sigma) supplemented with 10% charcoal/dextran‐treated foetal bovine serum (cFBS) (Biological Industries), 1.5 g/L sodium bicarbonate, 1% ITS‐G (Gibco), and 500 ng/mL puromycin. Ishikawa cells were also obtained from ATCC. Ishikawa cells were cultured in phenol red‐free DMEM/F12 medium supplemented with sodium pyruvate, penicillin/streptomycin, and 10% foetal bovine serum (FBS) (Biological Industries). AN3‐CA cells were cultured in Eagle's Minimum Essential Medium (Sigma) supplemented with 10% FBS, 2.2 g/L sodium bicarbonate, 0.292 g/L L‐glutamine, and penicillin/streptomycin. All cells were maintained in an incubator at 37°C under a 5% CO2 atmosphere. For further experiments, cultured cells were treated with ATP (sigma), suramin (100 μmol/L, Santa Cruz Biotechnology), apyrase (1 U/mL, Sigma), ARL67156 (200 μmol/L, Sigma), carbenoxolone (Sigma), IL8 recombinant protein (R&D), cathepsin B recombinant proteins (R&D), trypsin (Amerso) and lactate (Aladdin).
5
Immortalized Endometrial Stromal Cell Culture
6
Endometrial Biopsy and Decidualization Protocol
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Endometrial biopsies were obtained from 7 pre-menopausal women with regular menstrual cycles (5 in proliferative phase and 2 in secretory phase) and from 1 peri-menopause woman, all undergoing histeroscopy for diagnostic purposes. The selected women did not show any evident endometrial pathology or suffer from any endocrine disorder or systemic disease and have not received any steroid treatment for at least 3 months prior to tissue collection. Uterine samples were obtained by Vacuum Aspiration Biopsy Random Assay (Vabra) (see Supplementary Information for the details of tissue culture preparation).
The immortalized human endometrial stromal cell line T-HESC, originated from normal human endometrial stromal cell immortalized with hTERT27 (link), was obtained from ATCC (CRL-4003™). Cells were maintained in complete growth medium according to the manufacturer’s instructions. To induce T-HESC cell decidualization, stromal cells were stimulated with cAMP (0.5 mM; Sigma) and medroxyprogesterone acetate (MPA) (1 μM; Sigma) for 7 days.
The immortalized human endometrial stromal cell line T-HESC, originated from normal human endometrial stromal cell immortalized with hTERT27 (link), was obtained from ATCC (CRL-4003™). Cells were maintained in complete growth medium according to the manufacturer’s instructions. To induce T-HESC cell decidualization, stromal cells were stimulated with cAMP (0.5 mM; Sigma) and medroxyprogesterone acetate (MPA) (1 μM; Sigma) for 7 days.
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