Recombinant:Article Title: Modeling human embryo implantation in vitro.
Article Snippet: .. A 6 well plate was coated with 0.5 μg/mL Recombinant Laminin iMatrix511 (AMSbio, AMS.892 012) in 1.5 mL of TSC medium [DMEM/F12 supplemented with 0.1 mM 2-mercaptoethanol, 0.2% FBS, 0.5% Penicillin-Streptomycin, 0.3%, BSA (Gibco 15260-037), 1% ITS-X (Gibco, 51500), 1.5 μg/mL L-ascorbic acid (Sigma A4544), 50 ng/ml EGF (Peprotech AF-100-15), 2 μM CHIR99021 (Cambridge Stem Cell Institute), 0.5 μM A83-01, 1 μM SB431542 (Bio-Techne, 1614), 0.8 mM VPA (Sigma, P4543), and 5 μM Y-27632] at 37 ◦ C for 30mins. .. Fragments were enzymatically digested with 1 mg/ml collagenase type I (Sigma-Aldrich, SCR103) in phenol red-free DMEM (Sigma-Aldrich, D4947) and incubated overnight at 4 ◦ C. After incubation, samples were vertically oriented for 10 minutes, allowing separation of epithelial and stromal compartments according to different fragments size by gravity sedimentation.
Article Title: Lipid desaturation regulates the balance between self-renewal and differentiation in mouse blastocyst-derived stem cells
Article Snippet: .. TSCs in maintenance were cultivated as previously described [ ], on gelatin coated mitomycin inactivated MEFs in 30% TSC medium [RPMI 1640 (Gibco, USA) supplemented with 20% FBS (Gibco, USA), 1% non-essential amino acids (Gibco, USA), 1% L-glutamine (Biological Industries, USA), 1% sodium pyruvate (Biological Industries, USA), 1% pen/strep (Biological Industries, USA), 50 μM β-mercaptoethanol (Gibco, USA)] and 70% MEF conditioned medium (medium collected from mitomycin inactivated MEFs, containing the same supplements) freshly supplemented with 25 ng/mL of human recombinant FGF4 (prepared in house) and 1 μg/mL heparin (Sigma-Aldrich, USA). ..
Affinity Magnetic Separation:Article Title: Modeling human embryo implantation in vitro.
Article Snippet: .. A 6 well plate was coated with 0.5 μg/mL Recombinant Laminin iMatrix511 (AMSbio, AMS.892 012) in 1.5 mL of TSC medium [DMEM/F12 supplemented with 0.1 mM 2-mercaptoethanol, 0.2% FBS, 0.5% Penicillin-Streptomycin, 0.3%, BSA (Gibco 15260-037), 1% ITS-X (Gibco, 51500), 1.5 μg/mL L-ascorbic acid (Sigma A4544), 50 ng/ml EGF (Peprotech AF-100-15), 2 μM CHIR99021 (Cambridge Stem Cell Institute), 0.5 μM A83-01, 1 μM SB431542 (Bio-Techne, 1614), 0.8 mM VPA (Sigma, P4543), and 5 μM Y-27632] at 37 ◦ C for 30mins. .. Fragments were enzymatically digested with 1 mg/ml collagenase type I (Sigma-Aldrich, SCR103) in phenol red-free DMEM (Sigma-Aldrich, D4947) and incubated overnight at 4 ◦ C. After incubation, samples were vertically oriented for 10 minutes, allowing separation of epithelial and stromal compartments according to different fragments size by gravity sedimentation.
Cell Culture:Article Title: Trophoblast stem cell derivation from naive and primed hPSC enables ELF5 functional analysis.
Article Snippet: .. TSC were cultured in plates coated with 5 μg/mL CollagenIV (Col-IV, Corning) in TSC medium (basal DMEM/F12 + GlutaMax supplemented with 0.1 mM 2-mercaptoethanol [Gibco], 0.2% fetal bovine serum [Gibco], 0.5% penicillinstreptomycin [Sigma-Aldrich, P4333], 0.3% BSA [Sigma, A7030-500G], 1% insulin-transferrin-selenium-ethanolamine [Gibco, 51500056], 1.5 μg/mL L-ascorbic acid [Sigma, A4544], 50 ng/mL EGF [Peprotech, AF-100-151MG], 2 μM CHIR99021 [Stemgent, 04-0004-10], 0.5 μM A83-01 [Sigma, SML0788-5MG], 1 μM SB431542 [Cayman Chemicals, 13031], 0.8 mM sodium butyrate [Sigma, (D) ELF5 CRISPR-Cas9 knockout guides targeting exon 2 common for all protein coding isoforms. (E) Immunocytochemistry of ELF5 in TSCWT and TSCKO lines. ..
Article Title: Trophoblast stem cell derivation from naive and primed hPSC enables ELF5 functional analysis
Article Snippet: .. TSC were cultured in plates coated with 5 μg/mL Collagen-IV (Col-IV, Corning) in TSC medium (basal DMEM/F12 + GlutaMax supplemented with 0.1 mM 2-mercaptoethanol [Gibco], 0.2% fetal bovine serum [Gibco], 0.5% penicillin-streptomycin [Sigma-Aldrich, P4333], 0.3% BSA [Sigma, A7030-500G], 1% insulin-transferrin-selenium-ethanolamine [Gibco, 51500056], 1.5 μg/mL L-ascorbic acid [Sigma, A4544], 50 ng/mL EGF [Peprotech, AF-100-15-1MG], 2 μM CHIR99021 [Stemgent, 04-0004-10], 0.5 μM A83-01 [Sigma, SML0788-5MG], 1 μM SB431542 [Cayman Chemicals, 13031], 0.8 mM sodium butyrate [Sigma, B5887-1G], and 5 μM Y-27632 [Selleckchem, S1049]). .. ESCp and iPSC were cultured on wells coated with Matrigel (Corning) in E8 (Gibco) or E8 Flex (Gibco) medium.
CRISPR:Article Title: Trophoblast stem cell derivation from naive and primed hPSC enables ELF5 functional analysis.
Article Snippet: .. TSC were cultured in plates coated with 5 μg/mL CollagenIV (Col-IV, Corning) in TSC medium (basal DMEM/F12 + GlutaMax supplemented with 0.1 mM 2-mercaptoethanol [Gibco], 0.2% fetal bovine serum [Gibco], 0.5% penicillinstreptomycin [Sigma-Aldrich, P4333], 0.3% BSA [Sigma, A7030-500G], 1% insulin-transferrin-selenium-ethanolamine [Gibco, 51500056], 1.5 μg/mL L-ascorbic acid [Sigma, A4544], 50 ng/mL EGF [Peprotech, AF-100-151MG], 2 μM CHIR99021 [Stemgent, 04-0004-10], 0.5 μM A83-01 [Sigma, SML0788-5MG], 1 μM SB431542 [Cayman Chemicals, 13031], 0.8 mM sodium butyrate [Sigma, (D) ELF5 CRISPR-Cas9 knockout guides targeting exon 2 common for all protein coding isoforms. (E) Immunocytochemistry of ELF5 in TSCWT and TSCKO lines. ..
Knock-Out:Article Title: Trophoblast stem cell derivation from naive and primed hPSC enables ELF5 functional analysis.
Article Snippet: .. TSC were cultured in plates coated with 5 μg/mL CollagenIV (Col-IV, Corning) in TSC medium (basal DMEM/F12 + GlutaMax supplemented with 0.1 mM 2-mercaptoethanol [Gibco], 0.2% fetal bovine serum [Gibco], 0.5% penicillinstreptomycin [Sigma-Aldrich, P4333], 0.3% BSA [Sigma, A7030-500G], 1% insulin-transferrin-selenium-ethanolamine [Gibco, 51500056], 1.5 μg/mL L-ascorbic acid [Sigma, A4544], 50 ng/mL EGF [Peprotech, AF-100-151MG], 2 μM CHIR99021 [Stemgent, 04-0004-10], 0.5 μM A83-01 [Sigma, SML0788-5MG], 1 μM SB431542 [Cayman Chemicals, 13031], 0.8 mM sodium butyrate [Sigma, (D) ELF5 CRISPR-Cas9 knockout guides targeting exon 2 common for all protein coding isoforms. (E) Immunocytochemistry of ELF5 in TSCWT and TSCKO lines. ..
Immunocytochemistry:Article Title: Trophoblast stem cell derivation from naive and primed hPSC enables ELF5 functional analysis.
Article Snippet: .. TSC were cultured in plates coated with 5 μg/mL CollagenIV (Col-IV, Corning) in TSC medium (basal DMEM/F12 + GlutaMax supplemented with 0.1 mM 2-mercaptoethanol [Gibco], 0.2% fetal bovine serum [Gibco], 0.5% penicillinstreptomycin [Sigma-Aldrich, P4333], 0.3% BSA [Sigma, A7030-500G], 1% insulin-transferrin-selenium-ethanolamine [Gibco, 51500056], 1.5 μg/mL L-ascorbic acid [Sigma, A4544], 50 ng/mL EGF [Peprotech, AF-100-151MG], 2 μM CHIR99021 [Stemgent, 04-0004-10], 0.5 μM A83-01 [Sigma, SML0788-5MG], 1 μM SB431542 [Cayman Chemicals, 13031], 0.8 mM sodium butyrate [Sigma, (D) ELF5 CRISPR-Cas9 knockout guides targeting exon 2 common for all protein coding isoforms. (E) Immunocytochemistry of ELF5 in TSCWT and TSCKO lines. ..
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