Recombinant:Article Title: A molecular cell atlas of endocrine signaling in human neural organoids
Article Snippet: .. REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Anti-TUBB3 Biolegend Cat# 801201; RRID:AB_2313773 Anti-MAP2 Synaptic System Cat #188004; RRID:AB_2138181 Anti-SOX2 R&D Systems Cat #AF2018; RRID:AB_355110 Anti-Ki-67 Abcam Cat #Ab15580; RRID:AB_443209 Chemicals, peptides, and recombinant proteins Fulvestrant Sigma-Aldrich Cat #I4409 Estradiol Sigma-Aldrich Cat #E8875 NH3 Medchem Express Cat #HY-141513 T3 Sigma-Aldrich Cat #T2877 RU38386 Sigma-Aldrich Cat #M8046 Dexamethasone Sigma-Aldrich Cat #D4902 AGN193109 Sigma-Aldrich Cat #SML2034 ATRA Sigma-Aldrich Cat #R2625 3-methoxy-4-nitroflavone Clearsynth Cat #CS-T-79815 Benzo(a)pyrene Sigma-Aldrich Cat #B1760 Hydroxyflutamide Sigma-Aldrich Cat #H4166 Testosterone Sigma-Aldrich Cat #86500 SR9238 Sigma-Aldrich Cat #G6295 GW3965 Sigma-Aldrich Cat #SML1510 Countess cell counting chamber slides Thermo Fischer Scientific Cat #C10283 Trypan Blue Gibco Cat #15250062 TeSR/E8 medium Stemcell Technologies Cat #05990 ReLeSR Stemcell Technologies Cat#05872 Accutase Sigma-Aldrich Cat#A6964 Y-27632 Tocris Cat1254 DMEM/F12 medium Gibco Cat #11330057 Knockout serum Gibco Cat #10828028 Non-essential amino acids Sigma-Aldrich Cat# M7145 2-mercaptoethanol Gibco Cat#31350010 GlutaMax Gibco Cat#35050061 Dorsomorphin Sigma-Aldrich Cat#P5499 SB431542 Medchem Express Cat# HY-10431 neurobasal medium Gibco Cat#12348017 B-27 Supplement minus Vitamin A Gibco Cat# 12587001 FGF2 Prepro Tech Cat# 100-18B EGF Prepro Tech Cat# AF-100-15 BDNF Prepro Tech Cat# 450- 02 Neurotrophin 3 Prepro Tech Cat# 450-03 Matrigel Corning Cat#354277 4% paraformaldehyde/PBS solution SantaCruz Cat# sc-261692 Sodium citrate buffer Ph.6 WVR chemicals Cat#27833.294P Normal donkey serum Jackson ImmunoResearch Cat#017-000-121 Triton X-100 Sigma-Aldrich Cat# T8787 RNAse-free DNase Qiagen Cat#79265 (Continued on next page) Cell Press Blue 1, 100077, August 17, 2026 e1 .. REAGENT or RESOURCE SOURCE IDENTIFIER Papain Stemcell Technologies Cat# 07466 Ovomucoid Worthington Biochemical Cat# LS003087 Penicillin and streptomycin Thermo Fischer Scientific Cat# 15140122 DAPI (4’,6-Diamidino-2-Phenylindole, Dilactate) BioLegend Cat#422801 DPBS w/o Calcium w/o Magnesium Euroclone Cat#ECB4004L Xilene Sigma-Aldrich Cat#534056 ProLongTM Gold Antifade Mountant Invitrogen Cat #P36930 Critical commercial assays PureLink RNA Mini kit Thermo Fischer Scientific Cat#12183025 High Sensitivity RNA ScreenTape Analysis Agilent Cat# 5067-5579 3’ CellPlex Kit Set A 10× Genomics Cat # 1000261 Chromium Next GEM Single Cell 3ʹ Reagent Kits v3.1 10× Genomics Cat#CG000315 Quick StartTM Bradford Protein Assay Kit BioRad Cat# 5000201 Deposited data Bulk RNA-seq dataset 1 BrainSpan http://www.brainspan.org/static/ download.html Bulk RNA-seq dataset 2 Cheroni, 2022 ArrayExpress: E-MTAB-11239 ScRNA-seq dataset 1 He,2024 https://cellxgene.cziscience.com/ collections/de379e5f-52d0-498c-98010f850823c847 ScRNA-seq dataset 2 Wang,2025 https://cellxgene.cziscience.com/ collections/ad2149fc-19c5-41de-8cfe44710fbada73 ScRNA-seq dataset 3 Caporale,2024 ArrayExpress: E-MTAB-14574 Neural organoids Bulk RNA-seq This study ArrayExpress: E-MTAB-17185 Neural organoids scRNA-seq This study ArrayExpress: E-MTAB-17184 Experimental models: Cell lines WTSli018-B-1 (CTL08) Wellcome Trust Institute Male hiPSC UMILi024-A (CTL04) Reprogrammed in house Female hiPSC Software and algorithms Original code This study GitHub: https://github.com/ GiuseppeTestaLab/noha, Zenodo: https:// doi.org/10.5281/zenodo.206146645 ScRNA-seq container analysis This study docker pull alessiavalenti/sc:sc-noha-0.0.1 BulkRNA-seq and steroidomics container analysis This study docker pull testalab/ downstream:EndPoints-1.1.5 Imaging container analysis This study docker pull alessiavalenti/ imaging:TMA-0.0.1 Other NanoDrop One Thermo Fischer Scientific Cat # 840274100 Countess 3 FL Thermo Fischer Scientific Cat#AMQAF2000 4200 TapeStation System Agilent Cat #G2991BA ZEISS Axio Scan.Z1 Zeiss N/A Chromium X 10× Genomics Cat #1000326 Triple Quad 6500+ mass spectrometer SCIEX Cat# Triple Quad 6500+ NovaSeq 6000 sequencer Illumina Cat #20012850 e2 Cell Press Blue 1, 100077, August 17, 2026
Article Title: Molecular and Proteomic Profiling of Human Cerebral Organoid‑Derived Small Extracellular Vesicles
Article Snippet: .. Briefly, hiPSCs were plated on a low‐attachment U‐bottom 96 well plate with embryonic stem cell media (ESCM) (DMEM/F‐12 (Gibco, Thermo Fisher Scientific, 31330‐038), 20% KnockOut Serum Replacement (KOSR) (Gibco, Thermo Fisher Scientific, 10828028), 1× MEM Non‐Essential Amino Acids Solution (MEM‐NEAA) (Lonza, 11140050), 1× GlutaMAX Supplement (Gibco, Thermo Fisher Scientific, 35050061), 1% Penicillin‐Streptomycin (Gibco, Thermo Fisher Scientific, 15140122), 0.385 mM 2‐mercaptoethanol (Gibco, Thermo Fisher Scientific, 21985023), Y‐27632 (50 μM) (Tocris, 1254), 4 ng/mL Human FGF‐basic Recombinant Protein (only for the first 4 days) (Gibco, Thermo Fisher Scientific, Cat no. PHG0024) and incubated with medium change every other day for the generation of the embryoid bodies (EBs). .. On day 6, EBs were transferred into the low attachment 12 well plates with Neural Induction Media (NIM) (DMEM/F‐12) supplemented with 1× N‐2 MAX Media Supplement (R&D Systems, AR009), 1× GlutaMAX, 1× MEM‐NEAA, 1 μg/mL Heparin (Sigma‐Aldrich, H3149) for 4 days.
Cell Counting:Article Title: A molecular cell atlas of endocrine signaling in human neural organoids
Article Snippet: .. REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Anti-TUBB3 Biolegend Cat# 801201; RRID:AB_2313773 Anti-MAP2 Synaptic System Cat #188004; RRID:AB_2138181 Anti-SOX2 R&D Systems Cat #AF2018; RRID:AB_355110 Anti-Ki-67 Abcam Cat #Ab15580; RRID:AB_443209 Chemicals, peptides, and recombinant proteins Fulvestrant Sigma-Aldrich Cat #I4409 Estradiol Sigma-Aldrich Cat #E8875 NH3 Medchem Express Cat #HY-141513 T3 Sigma-Aldrich Cat #T2877 RU38386 Sigma-Aldrich Cat #M8046 Dexamethasone Sigma-Aldrich Cat #D4902 AGN193109 Sigma-Aldrich Cat #SML2034 ATRA Sigma-Aldrich Cat #R2625 3-methoxy-4-nitroflavone Clearsynth Cat #CS-T-79815 Benzo(a)pyrene Sigma-Aldrich Cat #B1760 Hydroxyflutamide Sigma-Aldrich Cat #H4166 Testosterone Sigma-Aldrich Cat #86500 SR9238 Sigma-Aldrich Cat #G6295 GW3965 Sigma-Aldrich Cat #SML1510 Countess cell counting chamber slides Thermo Fischer Scientific Cat #C10283 Trypan Blue Gibco Cat #15250062 TeSR/E8 medium Stemcell Technologies Cat #05990 ReLeSR Stemcell Technologies Cat#05872 Accutase Sigma-Aldrich Cat#A6964 Y-27632 Tocris Cat1254 DMEM/F12 medium Gibco Cat #11330057 Knockout serum Gibco Cat #10828028 Non-essential amino acids Sigma-Aldrich Cat# M7145 2-mercaptoethanol Gibco Cat#31350010 GlutaMax Gibco Cat#35050061 Dorsomorphin Sigma-Aldrich Cat#P5499 SB431542 Medchem Express Cat# HY-10431 neurobasal medium Gibco Cat#12348017 B-27 Supplement minus Vitamin A Gibco Cat# 12587001 FGF2 Prepro Tech Cat# 100-18B EGF Prepro Tech Cat# AF-100-15 BDNF Prepro Tech Cat# 450- 02 Neurotrophin 3 Prepro Tech Cat# 450-03 Matrigel Corning Cat#354277 4% paraformaldehyde/PBS solution SantaCruz Cat# sc-261692 Sodium citrate buffer Ph.6 WVR chemicals Cat#27833.294P Normal donkey serum Jackson ImmunoResearch Cat#017-000-121 Triton X-100 Sigma-Aldrich Cat# T8787 RNAse-free DNase Qiagen Cat#79265 (Continued on next page) Cell Press Blue 1, 100077, August 17, 2026 e1 .. REAGENT or RESOURCE SOURCE IDENTIFIER Papain Stemcell Technologies Cat# 07466 Ovomucoid Worthington Biochemical Cat# LS003087 Penicillin and streptomycin Thermo Fischer Scientific Cat# 15140122 DAPI (4’,6-Diamidino-2-Phenylindole, Dilactate) BioLegend Cat#422801 DPBS w/o Calcium w/o Magnesium Euroclone Cat#ECB4004L Xilene Sigma-Aldrich Cat#534056 ProLongTM Gold Antifade Mountant Invitrogen Cat #P36930 Critical commercial assays PureLink RNA Mini kit Thermo Fischer Scientific Cat#12183025 High Sensitivity RNA ScreenTape Analysis Agilent Cat# 5067-5579 3’ CellPlex Kit Set A 10× Genomics Cat # 1000261 Chromium Next GEM Single Cell 3ʹ Reagent Kits v3.1 10× Genomics Cat#CG000315 Quick StartTM Bradford Protein Assay Kit BioRad Cat# 5000201 Deposited data Bulk RNA-seq dataset 1 BrainSpan http://www.brainspan.org/static/ download.html Bulk RNA-seq dataset 2 Cheroni, 2022 ArrayExpress: E-MTAB-11239 ScRNA-seq dataset 1 He,2024 https://cellxgene.cziscience.com/ collections/de379e5f-52d0-498c-98010f850823c847 ScRNA-seq dataset 2 Wang,2025 https://cellxgene.cziscience.com/ collections/ad2149fc-19c5-41de-8cfe44710fbada73 ScRNA-seq dataset 3 Caporale,2024 ArrayExpress: E-MTAB-14574 Neural organoids Bulk RNA-seq This study ArrayExpress: E-MTAB-17185 Neural organoids scRNA-seq This study ArrayExpress: E-MTAB-17184 Experimental models: Cell lines WTSli018-B-1 (CTL08) Wellcome Trust Institute Male hiPSC UMILi024-A (CTL04) Reprogrammed in house Female hiPSC Software and algorithms Original code This study GitHub: https://github.com/ GiuseppeTestaLab/noha, Zenodo: https:// doi.org/10.5281/zenodo.206146645 ScRNA-seq container analysis This study docker pull alessiavalenti/sc:sc-noha-0.0.1 BulkRNA-seq and steroidomics container analysis This study docker pull testalab/ downstream:EndPoints-1.1.5 Imaging container analysis This study docker pull alessiavalenti/ imaging:TMA-0.0.1 Other NanoDrop One Thermo Fischer Scientific Cat # 840274100 Countess 3 FL Thermo Fischer Scientific Cat#AMQAF2000 4200 TapeStation System Agilent Cat #G2991BA ZEISS Axio Scan.Z1 Zeiss N/A Chromium X 10× Genomics Cat #1000326 Triple Quad 6500+ mass spectrometer SCIEX Cat# Triple Quad 6500+ NovaSeq 6000 sequencer Illumina Cat #20012850 e2 Cell Press Blue 1, 100077, August 17, 2026
Knock-Out:Article Title: A molecular cell atlas of endocrine signaling in human neural organoids
Article Snippet: .. REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Anti-TUBB3 Biolegend Cat# 801201; RRID:AB_2313773 Anti-MAP2 Synaptic System Cat #188004; RRID:AB_2138181 Anti-SOX2 R&D Systems Cat #AF2018; RRID:AB_355110 Anti-Ki-67 Abcam Cat #Ab15580; RRID:AB_443209 Chemicals, peptides, and recombinant proteins Fulvestrant Sigma-Aldrich Cat #I4409 Estradiol Sigma-Aldrich Cat #E8875 NH3 Medchem Express Cat #HY-141513 T3 Sigma-Aldrich Cat #T2877 RU38386 Sigma-Aldrich Cat #M8046 Dexamethasone Sigma-Aldrich Cat #D4902 AGN193109 Sigma-Aldrich Cat #SML2034 ATRA Sigma-Aldrich Cat #R2625 3-methoxy-4-nitroflavone Clearsynth Cat #CS-T-79815 Benzo(a)pyrene Sigma-Aldrich Cat #B1760 Hydroxyflutamide Sigma-Aldrich Cat #H4166 Testosterone Sigma-Aldrich Cat #86500 SR9238 Sigma-Aldrich Cat #G6295 GW3965 Sigma-Aldrich Cat #SML1510 Countess cell counting chamber slides Thermo Fischer Scientific Cat #C10283 Trypan Blue Gibco Cat #15250062 TeSR/E8 medium Stemcell Technologies Cat #05990 ReLeSR Stemcell Technologies Cat#05872 Accutase Sigma-Aldrich Cat#A6964 Y-27632 Tocris Cat1254 DMEM/F12 medium Gibco Cat #11330057 Knockout serum Gibco Cat #10828028 Non-essential amino acids Sigma-Aldrich Cat# M7145 2-mercaptoethanol Gibco Cat#31350010 GlutaMax Gibco Cat#35050061 Dorsomorphin Sigma-Aldrich Cat#P5499 SB431542 Medchem Express Cat# HY-10431 neurobasal medium Gibco Cat#12348017 B-27 Supplement minus Vitamin A Gibco Cat# 12587001 FGF2 Prepro Tech Cat# 100-18B EGF Prepro Tech Cat# AF-100-15 BDNF Prepro Tech Cat# 450- 02 Neurotrophin 3 Prepro Tech Cat# 450-03 Matrigel Corning Cat#354277 4% paraformaldehyde/PBS solution SantaCruz Cat# sc-261692 Sodium citrate buffer Ph.6 WVR chemicals Cat#27833.294P Normal donkey serum Jackson ImmunoResearch Cat#017-000-121 Triton X-100 Sigma-Aldrich Cat# T8787 RNAse-free DNase Qiagen Cat#79265 (Continued on next page) Cell Press Blue 1, 100077, August 17, 2026 e1 .. REAGENT or RESOURCE SOURCE IDENTIFIER Papain Stemcell Technologies Cat# 07466 Ovomucoid Worthington Biochemical Cat# LS003087 Penicillin and streptomycin Thermo Fischer Scientific Cat# 15140122 DAPI (4’,6-Diamidino-2-Phenylindole, Dilactate) BioLegend Cat#422801 DPBS w/o Calcium w/o Magnesium Euroclone Cat#ECB4004L Xilene Sigma-Aldrich Cat#534056 ProLongTM Gold Antifade Mountant Invitrogen Cat #P36930 Critical commercial assays PureLink RNA Mini kit Thermo Fischer Scientific Cat#12183025 High Sensitivity RNA ScreenTape Analysis Agilent Cat# 5067-5579 3’ CellPlex Kit Set A 10× Genomics Cat # 1000261 Chromium Next GEM Single Cell 3ʹ Reagent Kits v3.1 10× Genomics Cat#CG000315 Quick StartTM Bradford Protein Assay Kit BioRad Cat# 5000201 Deposited data Bulk RNA-seq dataset 1 BrainSpan http://www.brainspan.org/static/ download.html Bulk RNA-seq dataset 2 Cheroni, 2022 ArrayExpress: E-MTAB-11239 ScRNA-seq dataset 1 He,2024 https://cellxgene.cziscience.com/ collections/de379e5f-52d0-498c-98010f850823c847 ScRNA-seq dataset 2 Wang,2025 https://cellxgene.cziscience.com/ collections/ad2149fc-19c5-41de-8cfe44710fbada73 ScRNA-seq dataset 3 Caporale,2024 ArrayExpress: E-MTAB-14574 Neural organoids Bulk RNA-seq This study ArrayExpress: E-MTAB-17185 Neural organoids scRNA-seq This study ArrayExpress: E-MTAB-17184 Experimental models: Cell lines WTSli018-B-1 (CTL08) Wellcome Trust Institute Male hiPSC UMILi024-A (CTL04) Reprogrammed in house Female hiPSC Software and algorithms Original code This study GitHub: https://github.com/ GiuseppeTestaLab/noha, Zenodo: https:// doi.org/10.5281/zenodo.206146645 ScRNA-seq container analysis This study docker pull alessiavalenti/sc:sc-noha-0.0.1 BulkRNA-seq and steroidomics container analysis This study docker pull testalab/ downstream:EndPoints-1.1.5 Imaging container analysis This study docker pull alessiavalenti/ imaging:TMA-0.0.1 Other NanoDrop One Thermo Fischer Scientific Cat # 840274100 Countess 3 FL Thermo Fischer Scientific Cat#AMQAF2000 4200 TapeStation System Agilent Cat #G2991BA ZEISS Axio Scan.Z1 Zeiss N/A Chromium X 10× Genomics Cat #1000326 Triple Quad 6500+ mass spectrometer SCIEX Cat# Triple Quad 6500+ NovaSeq 6000 sequencer Illumina Cat #20012850 e2 Cell Press Blue 1, 100077, August 17, 2026
Article Title: Molecular and Proteomic Profiling of Human Cerebral Organoid‑Derived Small Extracellular Vesicles
Article Snippet: .. Briefly, hiPSCs were plated on a low‐attachment U‐bottom 96 well plate with embryonic stem cell media (ESCM) (DMEM/F‐12 (Gibco, Thermo Fisher Scientific, 31330‐038), 20% KnockOut Serum Replacement (KOSR) (Gibco, Thermo Fisher Scientific, 10828028), 1× MEM Non‐Essential Amino Acids Solution (MEM‐NEAA) (Lonza, 11140050), 1× GlutaMAX Supplement (Gibco, Thermo Fisher Scientific, 35050061), 1% Penicillin‐Streptomycin (Gibco, Thermo Fisher Scientific, 15140122), 0.385 mM 2‐mercaptoethanol (Gibco, Thermo Fisher Scientific, 21985023), Y‐27632 (50 μM) (Tocris, 1254), 4 ng/mL Human FGF‐basic Recombinant Protein (only for the first 4 days) (Gibco, Thermo Fisher Scientific, Cat no. PHG0024) and incubated with medium change every other day for the generation of the embryoid bodies (EBs). .. On day 6, EBs were transferred into the low attachment 12 well plates with Neural Induction Media (NIM) (DMEM/F‐12) supplemented with 1× N‐2 MAX Media Supplement (R&D Systems, AR009), 1× GlutaMAX, 1× MEM‐NEAA, 1 μg/mL Heparin (Sigma‐Aldrich, H3149) for 4 days.
Knockdown:Article Title: Long-read RNA sequencing identifies non-coding isoform switching as a regulator of cell fate
Article Snippet: The knock-down was induced by adding 1 μg/mL doxycycline hyclate (Sigma-Aldrich #D9891) and ESCs were harvested after 5 days for RNA extraction. .. For human SNHG26 knock-down, primed hESC lines were seeded at a density of 2.6 ×104 cells/cm2 on GeltrexTM-coated plates (hESCqualified, growth factor-reduced, ThermoFisher Scientific, #A1413302) inmTeSRTM1 medium (StemCell Technologies, #85850) supplemented with 10 μM Y-27632 (Tocris Bioscience #1254). .. The following day, medium was replaced by fresh mTeSRTM1 medium and cells were cotransfected with 500 ng of hyPBase transposase, 1 μg of PB-TetO-AIOdCas9-BFP-KRAB vector and 875 ng of control or four SNHG26-specific pooled sgRNAs using LipofectamineTM Stem Transfection Reagent (ThermoFisher Scientific #STEM00015) according to the manufacturer’s instructions.
Cell Culture:Article Title: The gene-regulatory evolution of the human skeleton.
Article Snippet: .. In brief, iPS cells were counted and seeded at 9,000 cells per well in triplicate onto Matrigel-coated 24-well plates (10 μl ml−1 in DMEM, at least 1 h at 37 °C) and cultured for 24 h in mTeSR1 (STEMCELL Technologies, 85850) supplemented with Y-27632 (Tocris, 1254, 10 μM) and penicillin–streptomycin (25 IU ml−1). ..
Stem Cell:Article Title: Molecular and Proteomic Profiling of Human Cerebral Organoid‑Derived Small Extracellular Vesicles
Article Snippet: .. Briefly, hiPSCs were plated on a low‐attachment U‐bottom 96 well plate with embryonic stem cell media (ESCM) (DMEM/F‐12 (Gibco, Thermo Fisher Scientific, 31330‐038), 20% KnockOut Serum Replacement (KOSR) (Gibco, Thermo Fisher Scientific, 10828028), 1× MEM Non‐Essential Amino Acids Solution (MEM‐NEAA) (Lonza, 11140050), 1× GlutaMAX Supplement (Gibco, Thermo Fisher Scientific, 35050061), 1% Penicillin‐Streptomycin (Gibco, Thermo Fisher Scientific, 15140122), 0.385 mM 2‐mercaptoethanol (Gibco, Thermo Fisher Scientific, 21985023), Y‐27632 (50 μM) (Tocris, 1254), 4 ng/mL Human FGF‐basic Recombinant Protein (only for the first 4 days) (Gibco, Thermo Fisher Scientific, Cat no. PHG0024) and incubated with medium change every other day for the generation of the embryoid bodies (EBs). .. On day 6, EBs were transferred into the low attachment 12 well plates with Neural Induction Media (NIM) (DMEM/F‐12) supplemented with 1× N‐2 MAX Media Supplement (R&D Systems, AR009), 1× GlutaMAX, 1× MEM‐NEAA, 1 μg/mL Heparin (Sigma‐Aldrich, H3149) for 4 days.
Incubation:Article Title: Molecular and Proteomic Profiling of Human Cerebral Organoid‑Derived Small Extracellular Vesicles
Article Snippet: .. Briefly, hiPSCs were plated on a low‐attachment U‐bottom 96 well plate with embryonic stem cell media (ESCM) (DMEM/F‐12 (Gibco, Thermo Fisher Scientific, 31330‐038), 20% KnockOut Serum Replacement (KOSR) (Gibco, Thermo Fisher Scientific, 10828028), 1× MEM Non‐Essential Amino Acids Solution (MEM‐NEAA) (Lonza, 11140050), 1× GlutaMAX Supplement (Gibco, Thermo Fisher Scientific, 35050061), 1% Penicillin‐Streptomycin (Gibco, Thermo Fisher Scientific, 15140122), 0.385 mM 2‐mercaptoethanol (Gibco, Thermo Fisher Scientific, 21985023), Y‐27632 (50 μM) (Tocris, 1254), 4 ng/mL Human FGF‐basic Recombinant Protein (only for the first 4 days) (Gibco, Thermo Fisher Scientific, Cat no. PHG0024) and incubated with medium change every other day for the generation of the embryoid bodies (EBs). .. On day 6, EBs were transferred into the low attachment 12 well plates with Neural Induction Media (NIM) (DMEM/F‐12) supplemented with 1× N‐2 MAX Media Supplement (R&D Systems, AR009), 1× GlutaMAX, 1× MEM‐NEAA, 1 μg/mL Heparin (Sigma‐Aldrich, H3149) for 4 days.
Centrifugation:Article Title: Retinal organoid–based evaluation of icariin-loaded extracellular vesicles for hypoxia-induced retinal injury via mitochondrial modulation
Article Snippet: Passaging was conducted every 4–5 days by treating the hESC colonies with Accutase (Stem Cell Technologies) at 37 °C for 5 min when 70%–80% confluency. .. Following the detachment of hESCs as single cells, they were resuspended in mTeSR1 supplemented with 20 μM Y-27632 (Tocris, Bristol, City of Bristol, United Kingdom), pelleted by centrifugation at 1,350 rpm for 10 min at room temperature, and subsequently passaged at a ratio 1:10 to 1:50. .. After passaging, the remaining cells are stored at −80 °C in a stock solution containing 90% KnockOut Serum Replacement (KOSR; Gibco, Waltham, MA, USA) and 10% dimethyl sulfoxide (DMSO; Duchefa Biochemie, Haarlem, North Holland, Netherlands).
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