mESCs (E14) were cultured in Dulbecco’s modified eagle medium (SH3002101; Hyclone, Chicago, IL, USA), supplemented with 15% fetal bovine serum (35-015-CV; Corning, New York, NY, USA), 1X penicillin-streptomycin (15140122; Gibco, Waltham, MA, USA), 1X GlutaMAX (35050061; Gibco, USA), 1 mM sodium pyruvate (11360070; Gibco, USA), 1X non-essential amino acid (11140050; Gibco, USA), 1X β-mercaptoethanol (21985023; Gibco, USA), and 106 U of leukemia inhibitory factor (ESG1107; Merck, Darmstadt, Germany), on gelatin-coated dishes. The initial EBs were formed in hanging drops (500 cells/drop) in either differentiation media containing GFs (30 ng/mL VEGF-A [293-VE-010; R&D systems, Minneapolis, MN, USA] and 50 ng/mL VEGF-C [2179-VC-025; R&D systems, USA]) (hereafter referred to as EBGF) or differentiation media containing 60 μL/mL of NPAC (hereafter referred to as EBNPAC). The released GF concentration from the NPAC was adjusted to reach ~30 ng/mL (VEGF-A) and ~50 ng/mL (VEGF-C) at differentiation day2 based on the study by La and Yang [15 (link)]. GF-free EB (hereafter referred to as EB) was also produced and used as the control group. Morphological analysis of EB was performed under an SEM. After 48 h of EB formation in hanging drops, 10 EBs/well were transferred to BioCoat Collagen I glass multi-well culture slide (354630; Corning, USA) and incubated for 8 d.
Esg1107
Manufactured by Merck Group
Sourced in United States, Germany
The ESG1107 is a laboratory equipment product manufactured by Merck Group. It is designed to perform a core function, but a detailed description cannot be provided while maintaining an unbiased and factual approach. More information may be available from Merck Group directly.
Automatically generated - may contain errors
Lab products found in correlation
Penicillin streptomycin, by Thermo Fisher Scientific (14 mentions)
Glutamax, by Thermo Fisher Scientific (12 mentions)
Non essential amino acids (neaa), by Thermo Fisher Scientific (9 mentions)
L glutamine, by Thermo Fisher Scientific (8 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (8 mentions)
β mercaptoethanol, by Thermo Fisher Scientific (7 mentions)
Knockout serum replacement (ksr), by Thermo Fisher Scientific (6 mentions)
Knockout dmem, by Thermo Fisher Scientific (6 mentions)
Sodium pyruvate, by Thermo Fisher Scientific (5 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (5 mentions)
Pd0325901, by Axon Medchem (4 mentions)
Chir99021, by Merck Group (4 mentions)
Tms 002 c, by Merck Group (3 mentions)
Tms 001 c, by Merck Group (3 mentions)
Scrc 1011, by American Type Culture Collection (3 mentions)
2 mercaptoethanol, by Thermo Fisher Scientific (3 mentions)
Mitomycin c, by Merck Group (3 mentions)
Pen strep, by Thermo Fisher Scientific (3 mentions)
M3148, by Merck Group (3 mentions)
Es 007 e, by Merck Group (2 mentions)
66 protocols using esg1107
1
Embryoid Body Formation with Controlled Growth Factors
2
Derivation and Maintenance of Mouse ESCs
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Mouse ESCs were cultured on mitomycin-C-treated mouse embryonic fibroblasts (MEFs) in six-well tissue culture plates (92006, TPP, Trasadingen, Switzerland). Mouse ESC maintenance medium was KnockOut™ Dulbecco’s modified Eagle medium [10829018, Thermo Fisher Scientific (Thermo)] supplemented with 20% HyClone™ fetal bovine plasma (SH30070.03, Cytiva), 1% 2-mercaptoethanol (ES-007-E, Merck), 1% nonessential amino acids (TMS-001-C, Merck), 1% nucleosides (ES-008-D, Merck), 1% L-glutamine (TMS-002-C, Merck), 100 U/mL penicillin-streptomycin (15140-122, Thermo), and 1,000 U/mL mouse leukemia inhibitory factor (ESG1107, Merck). Before use, 1 μM mirdametinib [PD0325901/162-25291, FUJIFILM Wako (Wako)] and 3 μM laduviglusib (CHIR-99021/TB4423-GMP, Bio-Techne) were added to mESC maintenance medium. Cells were passaged every 2 to 3 d.
For cryopreservation, CELLBANKER 1plus (CB023, Takara) was used. For thawing, 1 mL of phosphate-buffered saline (PBS, Wako, 048-29805) was added to a cryopreserved cell vial. Mouse ESCs were quickly dispersed by pipetting and transferred to 15-mL centrifuge tubes containing 9 mL of PBS. After centrifugation, the cells were suspended in mESC maintenance medium containing 1 μM PD0325901 and 3 μM CHIR-99021. The cell suspension was then seeded onto MEF feeder cells. The thawed mESCs were used for injection after more than one passage.
For cryopreservation, CELLBANKER 1plus (CB023, Takara) was used. For thawing, 1 mL of phosphate-buffered saline (PBS, Wako, 048-29805) was added to a cryopreserved cell vial. Mouse ESCs were quickly dispersed by pipetting and transferred to 15-mL centrifuge tubes containing 9 mL of PBS. After centrifugation, the cells were suspended in mESC maintenance medium containing 1 μM PD0325901 and 3 μM CHIR-99021. The cell suspension was then seeded onto MEF feeder cells. The thawed mESCs were used for injection after more than one passage.
3
Efficient Mouse Embryonic Germ Cell Culture
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
EGC was established as described previously42 (link). Briefly, sorted PGCs were cultured on Mitomycin C (Kyowa Kirin, Japan)-inactivated Sl/Sl4-m220 feeder cells43 (link) in DMEM (D6429, Sigma-Aldrich) supplemented with 15% KSR (10828010, Thermo Fisher Scientific), 1% Glutamax (35050061, Thermo Fisher Scientific), 1% non-essential amino acids (11140050, Thermo Fisher Scientific), 0.1 mM β-mercaptoethanol (198-15781, FUJIFILM Wako, Japan), 1000 unit/mL mLIF (ESG1107, Merck Millipore), and 12 ng/mL bFGF (450-33, PeproTech, NJ) until day 3. On day 3, the media was replaced with fresh media containing 1 μM PD325901 (04-0006, ReproCELL), 1 μM CHIR99021 (04-0004, ReproCELL, Japan) and 250 nM A83-01(039-24111, FUJIFILM Wako, Japan).
Cells were cultured under 20% O2 and 5% CO2 at 37 °C and the media were refreshed after 1, 3, and 5 days of culture. The efficiency of colony formation was determined after 7 days in culture as the number of colonies per seeded cell in a culture well.
Cells were cultured under 20% O2 and 5% CO2 at 37 °C and the media were refreshed after 1, 3, and 5 days of culture. The efficiency of colony formation was determined after 7 days in culture as the number of colonies per seeded cell in a culture well.
4
Maintenance of STO Feeder Cells and mESCs
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
STO feeder cells (ATCC, SCRC-1049) were maintained in 10-cm tissue culture dishes in culture medium consisting of Dulbecco’s Modified Eagle’s Medium (DMEM) (D6546, Sigma-Aldrich, St. Louis, MO, USA), 100 mL/L foetal bovine serum (FBS) (Gibco, 10270, Thermo Fisher Scientific, Waltham, MA, USA), 10 mL/L MEM non-essential amino acid (Sigma-Aldrich, M7145) and 10 mL/L l -glutamine (Sigma-Aldrich, G7513) at 37 °C in a humidified incubator with 5% CO2. Cells were passaged twice per week at a split ratio of 1:4–1:6 until reaching passage 15. E14-Bra-GFP mESC line used in this study was a generous gift from Georges Lacaud at the Paterson Institute for Cancer Research, Manchester. tdTomato-transduced E14-Bra-GFP mESCs were from our in-house stock. mESCs were maintained in the 6-well feeder plates in culture medium consisting of DMEM, 150 mL/L FBS (Sigma-Aldrich, F2442), 10 mL/L MEM non-essential amino acid, 10 mL/L l -glutamine, 0.1 mmol/L β-mercaptoethanol (Gibco, 31350) and 1000 U/mL mouse leukaemia inhibitory factor (mLIF) (ESG1107, Merck Millipore, Billerica, MA, USA) at 37 °C in a humidified incubator with 5% CO2. Cells were passaged every other day at a split ratio of 1:6–1:10 until reaching passage 40.
5
Culturing Mouse and Drosophila Cell Lines
6
Culturing Yeast, Caulobacter, and Mouse ESCs
7
Mouse PSC Culture and Passage
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The mouse PSCs (ATCC; SCRC-1011) were seeded at a density of 50 000 cells per cm2 on a monolayer of mouse embryonic fibroblasts (MEF), which were previously mitotically inactive for PSC growing. We used embryonic stem cell basal medium (ATCC; SCRR-2010) supplemented with 15% fetal bovine serum, 0.1 mM 2-mercaptoethanol (Invitrogen; 21985023), and 1000 U/ml mouse leukemia inhibitory factor (EMD Millipore; ESG1107). The culture dishes were incubated at 37 °C in a humidified 5% CO2 and 95% air incubator. When the cultures in colonies reached 70% confluency, doses of 5 × 105 PSCs were obtained and resuspended in 500 μl of isotonic salt solution (ISS).
8
Mouse Embryonic Stem Cell Culture
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The mouse embryonic SCs (mESCs) (ATCC, Manassas, VA, USA; SCRC-1011) were seeded at a density of 50,000 cells per cm2 on a monolayer of mouse embryonic fibroblasts which were previously mitotically inactive for mESC growing. We used mESC basal medium (ATCC, catalog number SCRR-2010) supplemented with 15% fetal bovine serum, 0.1 mM 2-mercaptoethanol (Invitrogen, Carlsbad, CA, USA; catalog number 21985023), and 1,000 U/mL mouse leukemia inhibitory factor (Chemicon, now part of EMD Millipore, Billerica, MA, USA; catalog number ESG1107). The culture dishes were incubated at 37°C in a humidified 5% CO2 and 95% air incubator [17 (link)]. When the cultures reached 70% confluency, doses of 20,000 and 50,000 mESCs were obtained and resuspended in 0.025 mL of 0.09% balanced salt solution (BSS).
9
Culturing Bra-GFP/Rosa26-E2C and GFP-KSCs
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Bra-GFP/Rosa26-E2C mESCs (Zhou et al., 2018 (link)) were maintained in 0.1% gelatinised six-well tissue culture plates with mitomycin-C (Sigma-Aldrich, M4287) inactivated STO (ATCC, SCRC-1049) feeder cells at 37°C in a humidified incubator with 5% CO2 in Dulbecco's modified Eagle's medium (DMEM) (Sigma-Aldrich, D6546) supplemented with 15% fetal bovine serum (FBS) (Sigma-Aldrich, F2442), 1% minimum essential medium (MEM) non-essential amino acid (Sigma-Aldrich, M7145), 2 mmol l−1 L-glutamine (Sigma-Aldrich, G7513), 0.1 mmol l−1 β-mercaptoethanol (Gibco, 31350) and 1000 U ml−1 mouse leukaemia inhibitory factor (mLIF) (Merck Millipore, ESG1107). Cells were passaged every other day and those at passage 13–22 were used for experiments.
GFP-expressing mouse neonatal kidney-derived stem cells (GFP-KSCs) (E. Ranghini, PhD thesis, 2011) were maintained in 60-mm tissue culture dishes at 37°C in a humidified incubator with 5% CO2 in DMEM supplemented with 10% FBS (Gibco, 10270), 1% MEM non-essential amino acid (Sigma-Aldrich, M7145), 2 mmol l−1 L-glutamine (Sigma-Aldrich, G7513) and 0.1 mmol l−1 β-mercaptoethanol (Gibco, 31350). Cells were passaged 2–3 times per week and those at passage 17–20 were used for experiments.
GFP-expressing mouse neonatal kidney-derived stem cells (GFP-KSCs) (E. Ranghini, PhD thesis, 2011) were maintained in 60-mm tissue culture dishes at 37°C in a humidified incubator with 5% CO2 in DMEM supplemented with 10% FBS (Gibco, 10270), 1% MEM non-essential amino acid (Sigma-Aldrich, M7145), 2 mmol l−1 L-glutamine (Sigma-Aldrich, G7513) and 0.1 mmol l−1 β-mercaptoethanol (Gibco, 31350). Cells were passaged 2–3 times per week and those at passage 17–20 were used for experiments.
10
Generating mESCs from WT and Morc3 Mutant Embryos
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
mESCs were generated from WT and Morc3MD41/MD41 preimplantation embryos and were cultured on 0.1% gelatin without feeders in mESC medium [Knockout DMEM (10829-018; Gibco), 10% FBS (DE14-801F; BioWhittaker), NEAA (11140; Gibco), l -Glutamine (25030-123; Gibco), Sodium Pyruvate (11360; Gibco), 2-Mercaptoethanol (31350; Gibco) and Leukemia Inhibitory Factor (ESG1107; Millipore)] plus MEK inhibitor PD0325901 (1 mM) and GSK3 inhibitor CHIR99021 (3 mM, Axon Medchem). Cell cultures tested negative for mycoplasma on a regular basis.
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!