human escs Search Results


human escs h9  (Inserm Transfert)
90
Inserm Transfert human escs h9
Human Escs H9, supplied by Inserm Transfert, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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human embryonic stem cell h9 total rna  (ScienCell)
90
ScienCell human embryonic stem cell h9 total rna
Human Embryonic Stem Cell H9 Total Rna, supplied by ScienCell, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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human embryonic stem cells (escs)  (Blackwell Verlag)
90
Blackwell Verlag human embryonic stem cells (escs)
Human Embryonic Stem Cells (Escs), supplied by Blackwell Verlag, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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human escs lines wa09  (Coriell Institute for Medical Research)
90
Coriell Institute for Medical Research human escs lines wa09
a, EB formation in the presence of DMSO, Y-27632 and CEPT. Human <t>ESCs</t> <t>(WA09)</t> cells were dissociated with Accutase and plated into 6-well ULA plates in E6 medium. Representative phase-contrast images were taken at 24 h post-plating. Scale bar, 50 μm. b, To generate single EBs, hESCs were dissociated with Accutase and plating into AggreWell plates (5,000 cells/well). Images were taken 24 h post-plating. In the presence of DMSO the vast majority cells underwent cell death and EB formation was not observed. Treatment with Y-27632 supported EB formation but significant numbers of dead cells were detected surrounding the EB. Note that CEPT enables EB formation without apparent cell death. Scale bar, 100 μm. c, Quantification of the diameter of single EBs (24 h post-plating). Data are mean ± s.d. (n = 20 EBs for Y-27632 and n = 22 for CEPT), **** P < 0.0001, two-tailed Student’s t -test. d, Single EB formation in 96-well ULA plates. Dissociated hESCs were plated into 96-well ULA plates at 2,000 cells/well. Live and dead cells were stained with calcein green AM and propidium iodide (PI) 24 h after cell seeding. Scale bars, 100 μm. e, Quantification of cell survival in single EBs at day 1 and day 7 by using the CTG 3D assay. Note the significant difference between Y-27632 and CEPT treatment at both timepoints. Data represent mean ± s.d. (n = 24 EBs for each group), **** P < 0.0001, one-way ANOVA. f, CEPT supports differentiation of single EBs into the three germ layers. Individual EBs were cultured in E6 medium to allow for spontaneous differentiation and analyzed on day 7 for the expression of PAX6, SOX17, and Brachyury using an optimized quantitative RT-PCR protocol that enabled detection of low transcript levels in single EBs (see Materials and Methods section for details). Data represent mean ± s.d. (n = 3 experiments and in each experiment 24 EBs were analyzed for each group), * P = 0.0327, two-tailed Student’s t -test. g, h, Cerebral organoids were generated by using Y-27632 and CEPT for the first 24 h. At day 30, organoids were fixed, sectioned, processed for histology (hematoxylin and eosin stain) and immunohistochemistry for FOXG1. Representative images show that CEPT treatment resulted in larger organoids and more abundant FOXG1-expressing cells. Scale bars, 400 μm.
Human Escs Lines Wa09, supplied by Coriell Institute for Medical Research, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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stepwise differentiation of human escs into definitive endoderm  (ViaCyte Inc)
90
ViaCyte Inc stepwise differentiation of human escs into definitive endoderm
a, EB formation in the presence of DMSO, Y-27632 and CEPT. Human <t>ESCs</t> <t>(WA09)</t> cells were dissociated with Accutase and plated into 6-well ULA plates in E6 medium. Representative phase-contrast images were taken at 24 h post-plating. Scale bar, 50 μm. b, To generate single EBs, hESCs were dissociated with Accutase and plating into AggreWell plates (5,000 cells/well). Images were taken 24 h post-plating. In the presence of DMSO the vast majority cells underwent cell death and EB formation was not observed. Treatment with Y-27632 supported EB formation but significant numbers of dead cells were detected surrounding the EB. Note that CEPT enables EB formation without apparent cell death. Scale bar, 100 μm. c, Quantification of the diameter of single EBs (24 h post-plating). Data are mean ± s.d. (n = 20 EBs for Y-27632 and n = 22 for CEPT), **** P < 0.0001, two-tailed Student’s t -test. d, Single EB formation in 96-well ULA plates. Dissociated hESCs were plated into 96-well ULA plates at 2,000 cells/well. Live and dead cells were stained with calcein green AM and propidium iodide (PI) 24 h after cell seeding. Scale bars, 100 μm. e, Quantification of cell survival in single EBs at day 1 and day 7 by using the CTG 3D assay. Note the significant difference between Y-27632 and CEPT treatment at both timepoints. Data represent mean ± s.d. (n = 24 EBs for each group), **** P < 0.0001, one-way ANOVA. f, CEPT supports differentiation of single EBs into the three germ layers. Individual EBs were cultured in E6 medium to allow for spontaneous differentiation and analyzed on day 7 for the expression of PAX6, SOX17, and Brachyury using an optimized quantitative RT-PCR protocol that enabled detection of low transcript levels in single EBs (see Materials and Methods section for details). Data represent mean ± s.d. (n = 3 experiments and in each experiment 24 EBs were analyzed for each group), * P = 0.0327, two-tailed Student’s t -test. g, h, Cerebral organoids were generated by using Y-27632 and CEPT for the first 24 h. At day 30, organoids were fixed, sectioned, processed for histology (hematoxylin and eosin stain) and immunohistochemistry for FOXG1. Representative images show that CEPT treatment resulted in larger organoids and more abundant FOXG1-expressing cells. Scale bars, 400 μm.
Stepwise Differentiation Of Human Escs Into Definitive Endoderm, supplied by ViaCyte Inc, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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human escs ucla6  (Stem Cell Research Center)
90
Stem Cell Research Center human escs ucla6
a, EB formation in the presence of DMSO, Y-27632 and CEPT. Human <t>ESCs</t> <t>(WA09)</t> cells were dissociated with Accutase and plated into 6-well ULA plates in E6 medium. Representative phase-contrast images were taken at 24 h post-plating. Scale bar, 50 μm. b, To generate single EBs, hESCs were dissociated with Accutase and plating into AggreWell plates (5,000 cells/well). Images were taken 24 h post-plating. In the presence of DMSO the vast majority cells underwent cell death and EB formation was not observed. Treatment with Y-27632 supported EB formation but significant numbers of dead cells were detected surrounding the EB. Note that CEPT enables EB formation without apparent cell death. Scale bar, 100 μm. c, Quantification of the diameter of single EBs (24 h post-plating). Data are mean ± s.d. (n = 20 EBs for Y-27632 and n = 22 for CEPT), **** P < 0.0001, two-tailed Student’s t -test. d, Single EB formation in 96-well ULA plates. Dissociated hESCs were plated into 96-well ULA plates at 2,000 cells/well. Live and dead cells were stained with calcein green AM and propidium iodide (PI) 24 h after cell seeding. Scale bars, 100 μm. e, Quantification of cell survival in single EBs at day 1 and day 7 by using the CTG 3D assay. Note the significant difference between Y-27632 and CEPT treatment at both timepoints. Data represent mean ± s.d. (n = 24 EBs for each group), **** P < 0.0001, one-way ANOVA. f, CEPT supports differentiation of single EBs into the three germ layers. Individual EBs were cultured in E6 medium to allow for spontaneous differentiation and analyzed on day 7 for the expression of PAX6, SOX17, and Brachyury using an optimized quantitative RT-PCR protocol that enabled detection of low transcript levels in single EBs (see Materials and Methods section for details). Data represent mean ± s.d. (n = 3 experiments and in each experiment 24 EBs were analyzed for each group), * P = 0.0327, two-tailed Student’s t -test. g, h, Cerebral organoids were generated by using Y-27632 and CEPT for the first 24 h. At day 30, organoids were fixed, sectioned, processed for histology (hematoxylin and eosin stain) and immunohistochemistry for FOXG1. Representative images show that CEPT treatment resulted in larger organoids and more abundant FOXG1-expressing cells. Scale bars, 400 μm.
Human Escs Ucla6, supplied by Stem Cell Research Center, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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human escs  (Avior Integrated Products)
90
Avior Integrated Products human escs
a, EB formation in the presence of DMSO, Y-27632 and CEPT. Human <t>ESCs</t> <t>(WA09)</t> cells were dissociated with Accutase and plated into 6-well ULA plates in E6 medium. Representative phase-contrast images were taken at 24 h post-plating. Scale bar, 50 μm. b, To generate single EBs, hESCs were dissociated with Accutase and plating into AggreWell plates (5,000 cells/well). Images were taken 24 h post-plating. In the presence of DMSO the vast majority cells underwent cell death and EB formation was not observed. Treatment with Y-27632 supported EB formation but significant numbers of dead cells were detected surrounding the EB. Note that CEPT enables EB formation without apparent cell death. Scale bar, 100 μm. c, Quantification of the diameter of single EBs (24 h post-plating). Data are mean ± s.d. (n = 20 EBs for Y-27632 and n = 22 for CEPT), **** P < 0.0001, two-tailed Student’s t -test. d, Single EB formation in 96-well ULA plates. Dissociated hESCs were plated into 96-well ULA plates at 2,000 cells/well. Live and dead cells were stained with calcein green AM and propidium iodide (PI) 24 h after cell seeding. Scale bars, 100 μm. e, Quantification of cell survival in single EBs at day 1 and day 7 by using the CTG 3D assay. Note the significant difference between Y-27632 and CEPT treatment at both timepoints. Data represent mean ± s.d. (n = 24 EBs for each group), **** P < 0.0001, one-way ANOVA. f, CEPT supports differentiation of single EBs into the three germ layers. Individual EBs were cultured in E6 medium to allow for spontaneous differentiation and analyzed on day 7 for the expression of PAX6, SOX17, and Brachyury using an optimized quantitative RT-PCR protocol that enabled detection of low transcript levels in single EBs (see Materials and Methods section for details). Data represent mean ± s.d. (n = 3 experiments and in each experiment 24 EBs were analyzed for each group), * P = 0.0327, two-tailed Student’s t -test. g, h, Cerebral organoids were generated by using Y-27632 and CEPT for the first 24 h. At day 30, organoids were fixed, sectioned, processed for histology (hematoxylin and eosin stain) and immunohistochemistry for FOXG1. Representative images show that CEPT treatment resulted in larger organoids and more abundant FOXG1-expressing cells. Scale bars, 400 μm.
Human Escs, supplied by Avior Integrated Products, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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human escs  (Stem Cell Research Center)
90
Stem Cell Research Center human escs
a, EB formation in the presence of DMSO, Y-27632 and CEPT. Human <t>ESCs</t> <t>(WA09)</t> cells were dissociated with Accutase and plated into 6-well ULA plates in E6 medium. Representative phase-contrast images were taken at 24 h post-plating. Scale bar, 50 μm. b, To generate single EBs, hESCs were dissociated with Accutase and plating into AggreWell plates (5,000 cells/well). Images were taken 24 h post-plating. In the presence of DMSO the vast majority cells underwent cell death and EB formation was not observed. Treatment with Y-27632 supported EB formation but significant numbers of dead cells were detected surrounding the EB. Note that CEPT enables EB formation without apparent cell death. Scale bar, 100 μm. c, Quantification of the diameter of single EBs (24 h post-plating). Data are mean ± s.d. (n = 20 EBs for Y-27632 and n = 22 for CEPT), **** P < 0.0001, two-tailed Student’s t -test. d, Single EB formation in 96-well ULA plates. Dissociated hESCs were plated into 96-well ULA plates at 2,000 cells/well. Live and dead cells were stained with calcein green AM and propidium iodide (PI) 24 h after cell seeding. Scale bars, 100 μm. e, Quantification of cell survival in single EBs at day 1 and day 7 by using the CTG 3D assay. Note the significant difference between Y-27632 and CEPT treatment at both timepoints. Data represent mean ± s.d. (n = 24 EBs for each group), **** P < 0.0001, one-way ANOVA. f, CEPT supports differentiation of single EBs into the three germ layers. Individual EBs were cultured in E6 medium to allow for spontaneous differentiation and analyzed on day 7 for the expression of PAX6, SOX17, and Brachyury using an optimized quantitative RT-PCR protocol that enabled detection of low transcript levels in single EBs (see Materials and Methods section for details). Data represent mean ± s.d. (n = 3 experiments and in each experiment 24 EBs were analyzed for each group), * P = 0.0327, two-tailed Student’s t -test. g, h, Cerebral organoids were generated by using Y-27632 and CEPT for the first 24 h. At day 30, organoids were fixed, sectioned, processed for histology (hematoxylin and eosin stain) and immunohistochemistry for FOXG1. Representative images show that CEPT treatment resulted in larger organoids and more abundant FOXG1-expressing cells. Scale bars, 400 μm.
Human Escs, supplied by Stem Cell Research Center, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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keep-cryopreserved human escs  (STEMCELL Technologies Inc)
90
STEMCELL Technologies Inc keep-cryopreserved human escs
a, EB formation in the presence of DMSO, Y-27632 and CEPT. Human <t>ESCs</t> <t>(WA09)</t> cells were dissociated with Accutase and plated into 6-well ULA plates in E6 medium. Representative phase-contrast images were taken at 24 h post-plating. Scale bar, 50 μm. b, To generate single EBs, hESCs were dissociated with Accutase and plating into AggreWell plates (5,000 cells/well). Images were taken 24 h post-plating. In the presence of DMSO the vast majority cells underwent cell death and EB formation was not observed. Treatment with Y-27632 supported EB formation but significant numbers of dead cells were detected surrounding the EB. Note that CEPT enables EB formation without apparent cell death. Scale bar, 100 μm. c, Quantification of the diameter of single EBs (24 h post-plating). Data are mean ± s.d. (n = 20 EBs for Y-27632 and n = 22 for CEPT), **** P < 0.0001, two-tailed Student’s t -test. d, Single EB formation in 96-well ULA plates. Dissociated hESCs were plated into 96-well ULA plates at 2,000 cells/well. Live and dead cells were stained with calcein green AM and propidium iodide (PI) 24 h after cell seeding. Scale bars, 100 μm. e, Quantification of cell survival in single EBs at day 1 and day 7 by using the CTG 3D assay. Note the significant difference between Y-27632 and CEPT treatment at both timepoints. Data represent mean ± s.d. (n = 24 EBs for each group), **** P < 0.0001, one-way ANOVA. f, CEPT supports differentiation of single EBs into the three germ layers. Individual EBs were cultured in E6 medium to allow for spontaneous differentiation and analyzed on day 7 for the expression of PAX6, SOX17, and Brachyury using an optimized quantitative RT-PCR protocol that enabled detection of low transcript levels in single EBs (see Materials and Methods section for details). Data represent mean ± s.d. (n = 3 experiments and in each experiment 24 EBs were analyzed for each group), * P = 0.0327, two-tailed Student’s t -test. g, h, Cerebral organoids were generated by using Y-27632 and CEPT for the first 24 h. At day 30, organoids were fixed, sectioned, processed for histology (hematoxylin and eosin stain) and immunohistochemistry for FOXG1. Representative images show that CEPT treatment resulted in larger organoids and more abundant FOXG1-expressing cells. Scale bars, 400 μm.
Keep Cryopreserved Human Escs, supplied by STEMCELL Technologies Inc, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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human escs  (Active Motif)
90
Active Motif human escs
a, EB formation in the presence of DMSO, Y-27632 and CEPT. Human <t>ESCs</t> <t>(WA09)</t> cells were dissociated with Accutase and plated into 6-well ULA plates in E6 medium. Representative phase-contrast images were taken at 24 h post-plating. Scale bar, 50 μm. b, To generate single EBs, hESCs were dissociated with Accutase and plating into AggreWell plates (5,000 cells/well). Images were taken 24 h post-plating. In the presence of DMSO the vast majority cells underwent cell death and EB formation was not observed. Treatment with Y-27632 supported EB formation but significant numbers of dead cells were detected surrounding the EB. Note that CEPT enables EB formation without apparent cell death. Scale bar, 100 μm. c, Quantification of the diameter of single EBs (24 h post-plating). Data are mean ± s.d. (n = 20 EBs for Y-27632 and n = 22 for CEPT), **** P < 0.0001, two-tailed Student’s t -test. d, Single EB formation in 96-well ULA plates. Dissociated hESCs were plated into 96-well ULA plates at 2,000 cells/well. Live and dead cells were stained with calcein green AM and propidium iodide (PI) 24 h after cell seeding. Scale bars, 100 μm. e, Quantification of cell survival in single EBs at day 1 and day 7 by using the CTG 3D assay. Note the significant difference between Y-27632 and CEPT treatment at both timepoints. Data represent mean ± s.d. (n = 24 EBs for each group), **** P < 0.0001, one-way ANOVA. f, CEPT supports differentiation of single EBs into the three germ layers. Individual EBs were cultured in E6 medium to allow for spontaneous differentiation and analyzed on day 7 for the expression of PAX6, SOX17, and Brachyury using an optimized quantitative RT-PCR protocol that enabled detection of low transcript levels in single EBs (see Materials and Methods section for details). Data represent mean ± s.d. (n = 3 experiments and in each experiment 24 EBs were analyzed for each group), * P = 0.0327, two-tailed Student’s t -test. g, h, Cerebral organoids were generated by using Y-27632 and CEPT for the first 24 h. At day 30, organoids were fixed, sectioned, processed for histology (hematoxylin and eosin stain) and immunohistochemistry for FOXG1. Representative images show that CEPT treatment resulted in larger organoids and more abundant FOXG1-expressing cells. Scale bars, 400 μm.
Human Escs, supplied by Active Motif, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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human escs - by Bioz Stars, 2026-03
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human escs  (KU Leuven)
90
KU Leuven human escs
a, EB formation in the presence of DMSO, Y-27632 and CEPT. Human <t>ESCs</t> <t>(WA09)</t> cells were dissociated with Accutase and plated into 6-well ULA plates in E6 medium. Representative phase-contrast images were taken at 24 h post-plating. Scale bar, 50 μm. b, To generate single EBs, hESCs were dissociated with Accutase and plating into AggreWell plates (5,000 cells/well). Images were taken 24 h post-plating. In the presence of DMSO the vast majority cells underwent cell death and EB formation was not observed. Treatment with Y-27632 supported EB formation but significant numbers of dead cells were detected surrounding the EB. Note that CEPT enables EB formation without apparent cell death. Scale bar, 100 μm. c, Quantification of the diameter of single EBs (24 h post-plating). Data are mean ± s.d. (n = 20 EBs for Y-27632 and n = 22 for CEPT), **** P < 0.0001, two-tailed Student’s t -test. d, Single EB formation in 96-well ULA plates. Dissociated hESCs were plated into 96-well ULA plates at 2,000 cells/well. Live and dead cells were stained with calcein green AM and propidium iodide (PI) 24 h after cell seeding. Scale bars, 100 μm. e, Quantification of cell survival in single EBs at day 1 and day 7 by using the CTG 3D assay. Note the significant difference between Y-27632 and CEPT treatment at both timepoints. Data represent mean ± s.d. (n = 24 EBs for each group), **** P < 0.0001, one-way ANOVA. f, CEPT supports differentiation of single EBs into the three germ layers. Individual EBs were cultured in E6 medium to allow for spontaneous differentiation and analyzed on day 7 for the expression of PAX6, SOX17, and Brachyury using an optimized quantitative RT-PCR protocol that enabled detection of low transcript levels in single EBs (see Materials and Methods section for details). Data represent mean ± s.d. (n = 3 experiments and in each experiment 24 EBs were analyzed for each group), * P = 0.0327, two-tailed Student’s t -test. g, h, Cerebral organoids were generated by using Y-27632 and CEPT for the first 24 h. At day 30, organoids were fixed, sectioned, processed for histology (hematoxylin and eosin stain) and immunohistochemistry for FOXG1. Representative images show that CEPT treatment resulted in larger organoids and more abundant FOXG1-expressing cells. Scale bars, 400 μm.
Human Escs, supplied by KU Leuven, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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human escs - by Bioz Stars, 2026-03
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h9 human escs  (ReproCELL)
90
ReproCELL h9 human escs
a, EB formation in the presence of DMSO, Y-27632 and CEPT. Human <t>ESCs</t> <t>(WA09)</t> cells were dissociated with Accutase and plated into 6-well ULA plates in E6 medium. Representative phase-contrast images were taken at 24 h post-plating. Scale bar, 50 μm. b, To generate single EBs, hESCs were dissociated with Accutase and plating into AggreWell plates (5,000 cells/well). Images were taken 24 h post-plating. In the presence of DMSO the vast majority cells underwent cell death and EB formation was not observed. Treatment with Y-27632 supported EB formation but significant numbers of dead cells were detected surrounding the EB. Note that CEPT enables EB formation without apparent cell death. Scale bar, 100 μm. c, Quantification of the diameter of single EBs (24 h post-plating). Data are mean ± s.d. (n = 20 EBs for Y-27632 and n = 22 for CEPT), **** P < 0.0001, two-tailed Student’s t -test. d, Single EB formation in 96-well ULA plates. Dissociated hESCs were plated into 96-well ULA plates at 2,000 cells/well. Live and dead cells were stained with calcein green AM and propidium iodide (PI) 24 h after cell seeding. Scale bars, 100 μm. e, Quantification of cell survival in single EBs at day 1 and day 7 by using the CTG 3D assay. Note the significant difference between Y-27632 and CEPT treatment at both timepoints. Data represent mean ± s.d. (n = 24 EBs for each group), **** P < 0.0001, one-way ANOVA. f, CEPT supports differentiation of single EBs into the three germ layers. Individual EBs were cultured in E6 medium to allow for spontaneous differentiation and analyzed on day 7 for the expression of PAX6, SOX17, and Brachyury using an optimized quantitative RT-PCR protocol that enabled detection of low transcript levels in single EBs (see Materials and Methods section for details). Data represent mean ± s.d. (n = 3 experiments and in each experiment 24 EBs were analyzed for each group), * P = 0.0327, two-tailed Student’s t -test. g, h, Cerebral organoids were generated by using Y-27632 and CEPT for the first 24 h. At day 30, organoids were fixed, sectioned, processed for histology (hematoxylin and eosin stain) and immunohistochemistry for FOXG1. Representative images show that CEPT treatment resulted in larger organoids and more abundant FOXG1-expressing cells. Scale bars, 400 μm.
H9 Human Escs, supplied by ReproCELL, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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Image Search Results


a, EB formation in the presence of DMSO, Y-27632 and CEPT. Human ESCs (WA09) cells were dissociated with Accutase and plated into 6-well ULA plates in E6 medium. Representative phase-contrast images were taken at 24 h post-plating. Scale bar, 50 μm. b, To generate single EBs, hESCs were dissociated with Accutase and plating into AggreWell plates (5,000 cells/well). Images were taken 24 h post-plating. In the presence of DMSO the vast majority cells underwent cell death and EB formation was not observed. Treatment with Y-27632 supported EB formation but significant numbers of dead cells were detected surrounding the EB. Note that CEPT enables EB formation without apparent cell death. Scale bar, 100 μm. c, Quantification of the diameter of single EBs (24 h post-plating). Data are mean ± s.d. (n = 20 EBs for Y-27632 and n = 22 for CEPT), **** P < 0.0001, two-tailed Student’s t -test. d, Single EB formation in 96-well ULA plates. Dissociated hESCs were plated into 96-well ULA plates at 2,000 cells/well. Live and dead cells were stained with calcein green AM and propidium iodide (PI) 24 h after cell seeding. Scale bars, 100 μm. e, Quantification of cell survival in single EBs at day 1 and day 7 by using the CTG 3D assay. Note the significant difference between Y-27632 and CEPT treatment at both timepoints. Data represent mean ± s.d. (n = 24 EBs for each group), **** P < 0.0001, one-way ANOVA. f, CEPT supports differentiation of single EBs into the three germ layers. Individual EBs were cultured in E6 medium to allow for spontaneous differentiation and analyzed on day 7 for the expression of PAX6, SOX17, and Brachyury using an optimized quantitative RT-PCR protocol that enabled detection of low transcript levels in single EBs (see Materials and Methods section for details). Data represent mean ± s.d. (n = 3 experiments and in each experiment 24 EBs were analyzed for each group), * P = 0.0327, two-tailed Student’s t -test. g, h, Cerebral organoids were generated by using Y-27632 and CEPT for the first 24 h. At day 30, organoids were fixed, sectioned, processed for histology (hematoxylin and eosin stain) and immunohistochemistry for FOXG1. Representative images show that CEPT treatment resulted in larger organoids and more abundant FOXG1-expressing cells. Scale bars, 400 μm.

Journal: bioRxiv

Article Title: A Versatile Polypharmacology Platform Promotes Cytoprotection and Viability of Human Pluripotent and Differentiated Cells

doi: 10.1101/815761

Figure Lengend Snippet: a, EB formation in the presence of DMSO, Y-27632 and CEPT. Human ESCs (WA09) cells were dissociated with Accutase and plated into 6-well ULA plates in E6 medium. Representative phase-contrast images were taken at 24 h post-plating. Scale bar, 50 μm. b, To generate single EBs, hESCs were dissociated with Accutase and plating into AggreWell plates (5,000 cells/well). Images were taken 24 h post-plating. In the presence of DMSO the vast majority cells underwent cell death and EB formation was not observed. Treatment with Y-27632 supported EB formation but significant numbers of dead cells were detected surrounding the EB. Note that CEPT enables EB formation without apparent cell death. Scale bar, 100 μm. c, Quantification of the diameter of single EBs (24 h post-plating). Data are mean ± s.d. (n = 20 EBs for Y-27632 and n = 22 for CEPT), **** P < 0.0001, two-tailed Student’s t -test. d, Single EB formation in 96-well ULA plates. Dissociated hESCs were plated into 96-well ULA plates at 2,000 cells/well. Live and dead cells were stained with calcein green AM and propidium iodide (PI) 24 h after cell seeding. Scale bars, 100 μm. e, Quantification of cell survival in single EBs at day 1 and day 7 by using the CTG 3D assay. Note the significant difference between Y-27632 and CEPT treatment at both timepoints. Data represent mean ± s.d. (n = 24 EBs for each group), **** P < 0.0001, one-way ANOVA. f, CEPT supports differentiation of single EBs into the three germ layers. Individual EBs were cultured in E6 medium to allow for spontaneous differentiation and analyzed on day 7 for the expression of PAX6, SOX17, and Brachyury using an optimized quantitative RT-PCR protocol that enabled detection of low transcript levels in single EBs (see Materials and Methods section for details). Data represent mean ± s.d. (n = 3 experiments and in each experiment 24 EBs were analyzed for each group), * P = 0.0327, two-tailed Student’s t -test. g, h, Cerebral organoids were generated by using Y-27632 and CEPT for the first 24 h. At day 30, organoids were fixed, sectioned, processed for histology (hematoxylin and eosin stain) and immunohistochemistry for FOXG1. Representative images show that CEPT treatment resulted in larger organoids and more abundant FOXG1-expressing cells. Scale bars, 400 μm.

Article Snippet: All human ESCs lines (WA01, WA07, WA09) and iPSCs (LiPSC-GR1.1 and the following lines from Coriell Institute for Medical Research: GM25256 , GM23476, GM2610 and AICS-0023 clone 20) were maintained under feeder-free condition using Essential 8 (E8) medium and vitronectin (VTN-N) (Thermo Fisher Scientific).

Techniques: Two Tailed Test, Staining, Cell Culture, Expressing, Quantitative RT-PCR, Generated, H&E Stain, Immunohistochemistry