Cell Culture:Article Title: Effects of Natural Progesterone and Synthetic Progestin on Germ Layer Gene Expression in a Human Embryoid Body Model
Article Snippet: Use of human embryonic stem cell lines was approved by Institutional Review Board of Institute of Reproductive Medicine and Population, Medical Research Center, Seoul National University (219932-201307-LR-10-01-1) and animal experiments were approved by Institutional Animal Care and Use Committee (IACUC) of Seoul National University Hospital (15-0016-S1A0) and all the animal experiments were performed according to the ethical guidelines of the Association for Assessment and Accreditation of Laboratory Animal Care (AAALAC). .. As previously reported [ ], undifferentiated human embryonic stem cell line, SNUhES3 (46, XY) was cultured on a mitotically inactivated STO (CRL-1503, ATCC, Manassas, VA, USA) feeder layer and passaged every 7 days by mechanical dissociation [ ] under stereo-microscope (Nikon, Tokyo, Japan). .. The hESCs culture medium was composed of DMEM/F12 (Invitrogen, Waltham, MA, USA), 20% knockout serum replacement (KO-SR, Invitrogen, Carlsbad, CA, USA), 1% nonessential amino acids (Invitrogen, Carlsbad, CA, USA), 50 μg/mL streptomycin (Invitrogen, Carlsbad, CA, USA), 50 U/mL penicillin (Invitrogen, Carlsbad, CA, USA), 0.1 mM β-mercaptoethanol (Sigma-Aldrich, St. Louis, MO, USA), and 4 ng/mL basic fibroblast growth factor (bFGF, Invitrogen, Carlsbad, CA, USA).
Article Title: Biological and RNA regulatory function of MOV10 in mammalian germ cells
Article Snippet: .. For cell culture, we isolated Thy1-positive cells from 6- to 8-day-old B6;129S-Gt (Rosa) 26Sor/J mice and cultured these on 12-well plates with mitotically inactivated STO (SIM mouse embryo-derived thioguanine and ouabain-resistant feeder, SNLP76/7-4, ATCC) feeder layers in a defined serum-free, consisting of minimal essential medium (MEMa, Life Technology) supplemented with 2% bovine serum albumin (BSA, Sigma-Aldrich, St. Louis, MO, USA), 20 ng/ml GDNF (R&D Systems, Minneapolis, MN, USA), 150 ng/ml GFRA1 (R&D Systems) and 1 ng/ml basic fibroblast growth factor (FGF2; BD Biosciences), 10 μg/ml transferrin (Sigma-Aldrich), 50 μM free fatty acid mixture (5.6 mM linolenic acid, 13.4 mM oleic acid, 2.8 mM palmitoleic acid, 35.6 mM linoleic acid, 31.0 mM palmitic acid, 76.9 mM stearic acid; all from Sigma-Aldrich), 30 nM Na 2 SeO 3 (Sigma-Aldrich), 2 mM L-glutamine (Life Technology), 50 μM 2-mercaptoethanol (Sigma-Aldrich), 5 μg/ml insulin (Sigma-Aldrich), 10 mM HEPES (Sigma-Aldrich), and 60 μM putrescine (Sigma-Aldrich). ..
Article Title: Red Ginseng Extract Facilitates the Early Differentiation of Human Embryonic Stem Cells into Mesendoderm Lineage
Article Snippet: .. Briefly, hESCs were cultured on mitotically inactivated STO (CRL-1503; ATCC, USA) feeder layer by mechanical dissociation. hESC culture medium consisted of DMEM/F12 (Invitrogen), 20% knockout serum replacement (KO-SR; Invitrogen), 1% nonessential amino acids (Invitrogen), 50 U/ml penicillin (Invitrogen), 50 μ g/ml streptomycin (Invitrogen), 0.1 mM β -mercaptoethanol (Sigma, USA), and 4 ng/ml basic fibroblast growth factor (bFGF; Invitrogen). .. For EB formation, undifferentiated hESCs were treated with Collagenase type IV (Invitrogen) for 30 min at 37°C.
Article Title: A long non-coding RNA interacts with Gfra1 and maintains survival of mouse spermatogonial stem cells
Article Snippet: Briefly, germ cells were isolated from 6- to 8-day-old C57BL/6 or B6;129S-Gt(Rosa) 26Sor/J mice (Jackson Laboratory, Bar Harbor, ME, USA), and Thy1-positive cells were enriched using magnetic-activated cell separation (Miltenyi Biotech, Bergisch Gladbach, Germany). .. Cells were plated at a density of 1.5 to 2 × 10 5 per well on 12-well plates with mitotically inactivated STO (SIM mouse embryo-derived thioguanine- and ouabain-resistant feeder, SNLP76/7-4, ATCC, Manassas, VA, USA) feeder layers and cultured in a defined serum-free medium consisting of minimal essential medium (MEMa, Life Technology, Carlsbad, CA, USA) supplemented with 2% bovine serum albumin (BSA, Sigma-Aldrich, St. Louis, MO, USA), 20 ng/ml GDNF (R&D Systems, Minneapolis, MN, USA), 150 ng/ml GFRA1 (R&D Systems) and 1 ng/ml basic fibroblast growth factor (FGF2; BD Biosciences, San Jose, CA, USA), 10 μ g/ml transferrin (Sigma-Aldrich), 50 μ M free fatty acid mixture (5.6 mM linolenic acid, 13.4 mM oleic acid, 2.8 mM palmitoleic acid, 35.6 mM linoleic acid, 31.0 mM palmitic acid, 76.9 mM stearic acid; all from Sigma-Aldrich), 30 nM Na 2 SeO 3 (Sigma-Aldrich), 2 mM l -glutamine (Life Technology), 50 μ M 2-mercaptoethanol (Sigma-Aldrich), 5 μ g/ml insulin (Sigma-Aldrich), 10 mM HEPES (Sigma-Aldrich) and 60 μ M putrescine (Sigma-Aldrich). ..
Isolation:Article Title: Biological and RNA regulatory function of MOV10 in mammalian germ cells
Article Snippet: .. For cell culture, we isolated Thy1-positive cells from 6- to 8-day-old B6;129S-Gt (Rosa) 26Sor/J mice and cultured these on 12-well plates with mitotically inactivated STO (SIM mouse embryo-derived thioguanine and ouabain-resistant feeder, SNLP76/7-4, ATCC) feeder layers in a defined serum-free, consisting of minimal essential medium (MEMa, Life Technology) supplemented with 2% bovine serum albumin (BSA, Sigma-Aldrich, St. Louis, MO, USA), 20 ng/ml GDNF (R&D Systems, Minneapolis, MN, USA), 150 ng/ml GFRA1 (R&D Systems) and 1 ng/ml basic fibroblast growth factor (FGF2; BD Biosciences), 10 μg/ml transferrin (Sigma-Aldrich), 50 μM free fatty acid mixture (5.6 mM linolenic acid, 13.4 mM oleic acid, 2.8 mM palmitoleic acid, 35.6 mM linoleic acid, 31.0 mM palmitic acid, 76.9 mM stearic acid; all from Sigma-Aldrich), 30 nM Na 2 SeO 3 (Sigma-Aldrich), 2 mM L-glutamine (Life Technology), 50 μM 2-mercaptoethanol (Sigma-Aldrich), 5 μg/ml insulin (Sigma-Aldrich), 10 mM HEPES (Sigma-Aldrich), and 60 μM putrescine (Sigma-Aldrich). ..
Knock-Out:Article Title: Red Ginseng Extract Facilitates the Early Differentiation of Human Embryonic Stem Cells into Mesendoderm Lineage
Article Snippet: .. Briefly, hESCs were cultured on mitotically inactivated STO (CRL-1503; ATCC, USA) feeder layer by mechanical dissociation. hESC culture medium consisted of DMEM/F12 (Invitrogen), 20% knockout serum replacement (KO-SR; Invitrogen), 1% nonessential amino acids (Invitrogen), 50 U/ml penicillin (Invitrogen), 50 μ g/ml streptomycin (Invitrogen), 0.1 mM β -mercaptoethanol (Sigma, USA), and 4 ng/ml basic fibroblast growth factor (bFGF; Invitrogen). .. For EB formation, undifferentiated hESCs were treated with Collagenase type IV (Invitrogen) for 30 min at 37°C.
Article Title: DNA methyltransferase-3–dependent nonrandom template segregation in differentiating embryonic stem cells
Article Snippet: .. In brief, R1 cells were maintained on mitomycin C (Sigma-Aldrich) inactivated STO (CRL-1503; ATCC) feeder cells and fed daily with DMEM high-glucose medium supplemented with 15% KnockOut serum replacement, 2 mM GlutaMAX-I supplement, 1 mM MEM sodium pyruvate, 0.1 mM MEM nonessential amino acids (all obtained from Invitrogen), 55 μM 2-mercaptoethanol, and 1,000 U/ml ESGRO leukemia inhibitory factor (LIF; EMD Millipore). ..
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