From f0cb3a0e2df10568b26e8b47bd77d03b9b11408a Mon Sep 17 00:00:00 2001 From: jazmingarcia-github Date: Fri, 31 May 2024 12:50:26 -0700 Subject: [PATCH 1/4] Updated Stem cell characteristics for AICS-126_cl41 --- data/cell_line_catalog.json | 240 ++++++++++++++++++------------------ 1 file changed, 118 insertions(+), 122 deletions(-) diff --git a/data/cell_line_catalog.json b/data/cell_line_catalog.json index e89d809..327855d 100644 --- a/data/cell_line_catalog.json +++ b/data/cell_line_catalog.json @@ -12165,64 +12165,65 @@ "Protocols_banking": "/data/pdfs/AICS_Banking_Guidelines_EXTERNAL_v1.3_200210.pdf", "Stranalysis_report": "/data/pdfs/WTC11_STR_analysis.pdf" }, -{ - "cell_line_id": "AICS-0126", - "new": "true", - "clone_number": "41", - "alleleCount": "bi", - "status": "Yes", - "subpaged_status": [ - { "id": "editingdesign", - "done": "true" - }, - { - "id":"genomiccharacterizations", - "done": "true" - }, - { - "id": "validation", - "done": "false" - }, - { - "id": "stemcellcharacteristics", - "done": "false" - }, - { - "id": "supplementaryinformation", - "done": "true" - }, - { - "id": "protocols", - "done": "true" - }, - { - "id": "stranalysis", - "done": "true" - } - ], - "Main_data_available": "true", - "Main_parent_line": "WTC", - "Main_gene_symbol": ["CDH5"], - "Main_gene_name": "cadherin 5", - "Main_protein": "Vascular endothelial (VE)-cadherin", - "Main_gene_id": [1003], - "Main_isoforms": [ - { - "gene_name": "CDH5", - "gene_isoforms": ["NM_001795.5","XM_047433470.1", "XM_047433469.1", "XM_047433471.1"] - } - ], - "Main_structure": "endothelial adherens junctions", - "Main_terminal_tagged": "C-terminus", - "Main_fluorescent_tag": "mEGFP", - "Main_linker": "N/A", - "Main_order_link": "https://www.coriell.org/0/Sections/Search/Sample_Detail.aspx?Ref=AICS-0126-041&Product=iPSC&PgId=166", - "Main_cofa": "https://www.coriell.org/0/PDF/Allen/ipsc/AICS-0126-041_CofA.pdf", - "Main_parental_order_link": "https://catalog.coriell.org/0/Sections/Search/Sample_Detail.aspx?Ref=GM25256&Product=CC", - "Main_donor_plasmid": "", - "Main_eu_hpsc_reg": "https://hpscreg.eu/cell-line/UCSFi001-A-66", + { + "cell_line_id": "AICS-0126", + "new": "true", + "clone_number": "41", + "alleleCount": "bi", + "status": "Yes", + "subpaged_status": [ + { + "id": "editingdesign", + "done": "true" + }, + { + "id": "genomiccharacterizations", + "done": "true" + }, + { + "id": "validation", + "done": "false" + }, + { + "id": "stemcellcharacteristics", + "done": "false" + }, + { + "id": "supplementaryinformation", + "done": "true" + }, + { + "id": "protocols", + "done": "true" + }, + { + "id": "stranalysis", + "done": "true" + } + ], + "Main_data_available": "true", + "Main_parent_line": "WTC", + "Main_gene_symbol": [ "CDH5" ], + "Main_gene_name": "cadherin 5", + "Main_protein": "Vascular endothelial (VE)-cadherin", + "Main_gene_id": [ 1003 ], + "Main_isoforms": [ + { + "gene_name": "CDH5", + "gene_isoforms": [ "NM_001795.5", "XM_047433470.1", "XM_047433469.1", "XM_047433471.1" ] + } + ], + "Main_structure": "endothelial adherens junctions", + "Main_terminal_tagged": "C-terminus", + "Main_fluorescent_tag": "mEGFP", + "Main_linker": "N/A", + "Main_order_link": "https://www.coriell.org/0/Sections/Search/Sample_Detail.aspx?Ref=AICS-0126-041&Product=iPSC&PgId=166", + "Main_cofa": "https://www.coriell.org/0/PDF/Allen/ipsc/AICS-0126-041_CofA.pdf", + "Main_parental_order_link": "https://catalog.coriell.org/0/Sections/Search/Sample_Detail.aspx?Ref=GM25256&Product=CC", + "Main_donor_plasmid": "", + "Main_eu_hpsc_reg": "https://hpscreg.eu/cell-line/UCSFi001-A-66", - "Main_media": [ + "Main_media": [ { "type": "image", "cell_line_id": "AICS-0126", @@ -12239,17 +12240,17 @@ "caption": "One vial of distribution lot was thawed (cells were treated with ROCK inhibitor for 24hrs post-thaw-refer to culture protocol). Cultures were observed daily. Colonies were imaged one and three days post-thaw using a Leica microscope at 4x and 10x magnification.", "link": "/images/AICS-0126/AICS-126_cl41_CDH5.jpg" } - ], - "EditingDesign_crRNA_seq": "GGAGGAGCTGCTGTATTAGG", - "EditingDesign_cas9": "Wildtype spCas9", - "EditingDesign_NCBI_isoform": "", - "EditingDesign_gene_figure": "/images/AICS-0126/CDH5_GFP.png", - "EditingDesign_gene_figure_caption": "Top: CDH5 locus showing 3 CDH5 isoforms; Bottom: Zoom in on mEGFP insertion site at CDH5 C-terminal exon.", - "GenomicCharacterization_junction_schematic": "/images/GenomicCharacterization_junction_schematic_mEGFP.png", - "GenomicCharacterization_tagged_allele_gel": "/images/AICS-0126/CDH5_cl41_full_allele.jpg", - "GenomicCharacterization_tagged_allele_gel_caption": "Junctional PCR primers complementary to sequences flanking the homology arms in the distal genome were used together to amplify the tagged allele. Gel shows untagged allele in an unedited control and tagged allele (larger band) from single PCR reaction. (1% Agarose gel with Thermo Fisher 1kb ladder). Clone 41 is available from Coriell.", + ], + "EditingDesign_crRNA_seq": "GGAGGAGCTGCTGTATTAGG", + "EditingDesign_cas9": "Wildtype spCas9", + "EditingDesign_NCBI_isoform": "", + "EditingDesign_gene_figure": "/images/AICS-0126/CDH5_GFP.png", + "EditingDesign_gene_figure_caption": "Top: CDH5 locus showing 3 CDH5 isoforms; Bottom: Zoom in on mEGFP insertion site at CDH5 C-terminal exon.", + "GenomicCharacterization_junction_schematic": "/images/GenomicCharacterization_junction_schematic_mEGFP.png", + "GenomicCharacterization_tagged_allele_gel": "/images/AICS-0126/CDH5_cl41_full_allele.jpg", + "GenomicCharacterization_tagged_allele_gel_caption": "Junctional PCR primers complementary to sequences flanking the homology arms in the distal genome were used together to amplify the tagged allele. Gel shows untagged allele in an unedited control and tagged allele (larger band) from single PCR reaction. (1% Agarose gel with Thermo Fisher 1kb ladder). Clone 41 is available from Coriell.", - "GenomicCharacterization_junction_table": [ + "GenomicCharacterization_junction_table": [ { "editedGene": "CDH5-mEGFP", "junction": "5'", @@ -12274,26 +12275,26 @@ "expected_size": "2528", "confirmed_sequence": "Yes" } - ], - "GenomicCharacterization_junction_table_caption": "PCR amplified 5', 3', WT, and full allele junctions. 5', 3', and WT junctions were Sanger sequenced to check for precise mEGFP insertion.", + ], + "GenomicCharacterization_junction_table_caption": "PCR amplified 5', 3', WT, and full allele junctions. 5', 3', and WT junctions were Sanger sequenced to check for precise mEGFP insertion.", - "GenomicCharacterization_ddpcr": [ + "GenomicCharacterization_ddpcr": [ { "gene_tag": "CDH5-mEGFP", "clone": "cl41", "fp_ratio": "1.07", "plasmid": "N/A" } - ], - "GenomicCharacterization_ddpcr_caption": "FP:RPP30 ratio from ddPCR assay; values = 0.5 +/- 0.1 indicate heterozygous clone, values = 1 +/- 0.1 indicate homozygous clone. RPP30 is known 2n reference gene.", - "GenomicCharacterization_offtargets": "", - "GenomicCharacterization_offtargets_caption": "", - "Validation_WesternBlot": "", - "Validation_WesternBlot_caption": "", - "Validation_Immunofluorescence": "", - "Validation_Immunofluorescence_caption": "", + ], + "GenomicCharacterization_ddpcr_caption": "FP:RPP30 ratio from ddPCR assay; values = 0.5 +/- 0.1 indicate heterozygous clone, values = 1 +/- 0.1 indicate homozygous clone. RPP30 is known 2n reference gene.", + "GenomicCharacterization_offtargets": "", + "GenomicCharacterization_offtargets_caption": "", + "Validation_WesternBlot": "", + "Validation_WesternBlot_caption": "", + "Validation_Immunofluorescence": "", + "Validation_Immunofluorescence_caption": "", - "StemCellCharacterization_pluripotency_analysis": [ + "StemCellCharacterization_pluripotency_analysis": [ { "Marker": "NANOG", "positiveCells": "98.1" @@ -12304,7 +12305,7 @@ }, { "Marker": "OCT4", - "positiveCells": "98.3" + "positiveCells": "96.9" }, { "Marker": "SSEA-1", @@ -12313,52 +12314,47 @@ { "Marker": "SSEA-4", "positiveCells": "99.9" + } + ], + "StemCellCharacterization_pluripotency_analysis_caption": "iPSCs were stained with directly conjugated antibodies from BD Biosciences, acquired using a CytoFLEX (Beckman Coulter), and analyzed using FlowJo software (Treestar, Inc.). Doublets were excluded, then marker-specific gates were set according to corresponding fluorescence-minus-one (FMO) controls.", + "StemCellCharacterization_karyotype": "/images/AICS-0126/AICS-126_cl41_CDH5_karyotype.jpg", + "StemCellCharacterization_karyotype_caption": "After cells banks were created, one vial was thawed and 30 G-banded metaphase cells were karyotyped.", + "StemCellCharacterization_trilineage": [ + { + "Germ_Layer": "Ectoderm", + "Marker": "PAX6", + "percent_positive_cells": 98.4 }, { - "Marker": "TRA-160", - "positiveCells": "100" + "Germ_Layer": "Endoderm", + "Marker": "SOX17", + "percent_positive_cells": 93.8 + }, + { + "Germ_Layer": "Mesoderm", + "Marker": "Brachyury", + "percent_positive_cells": 95.1 } - ], - "StemCellCharacterization_pluripotency_analysis_caption": "iPSCs were stained with directly conjugated antibodies from BD Biosciences, acquired using a FACSAria III Fusion (BD Biosciences), and analyzed using FlowJo software (Treestar, Inc.). Doublets were excluded, then marker-specific gates were set according to corresponding fluorescence-minus-one (FMO) controls.", - "StemCellCharacterization_karyotype": "/images/AICS-0126/AICS-126_cl41_CDH5_karyotype.jpg", - "StemCellCharacterization_karyotype_caption": "After cells banks were created, one vial was thawed and 30 G-banded metaphase cells were karyotyped.", - "StemCellCharacterization_trilineage": [ - { - "Germ_Layer": "Ectoderm", - "Marker": "PAX6", - "percent_positive_cells": 98.4 - }, - { - "Germ_Layer": "Endoderm", - "Marker": "SOX17", - "percent_positive_cells": 93.8 - }, - { - "Germ_Layer": "Mesoderm", - "Marker": "Brachyury", - "percent_positive_cells": 95.1 - } - ], - "StemCellCharacterization_trilineage_caption": "iPSCs were subjected to a 5-7 day, non-terminal, directed differentiation using the STEMdiff™ Trilineage Differentiation Kit (STEMCELL Technologies, Inc.). Total RNA was isolated from each lineage specific differentiation and assayed via ddPCR for the expression of lineage specific transcripts; Pax6(Ectoderm), Sox17(Endoderm) and Brachyury(Mesoderm).", + ], + "StemCellCharacterization_trilineage_caption": "iPSCs were subjected to a 5-7 day, non-terminal, directed differentiation using the STEMdiff™ Trilineage Differentiation Kit (STEMCELL Technologies, Inc.). Total RNA was isolated from each lineage specific differentiation and assayed via ddPCR for the expression of lineage specific transcripts; Pax6(Ectoderm), Sox17(Endoderm) and Brachyury(Mesoderm).", - "StemCellCharacterization_differentiation": - { - "troponin_percent_positive": "65.2", - "Day_of_beating_percent": "100 (4)", - "Day_of_beating_range": "D8 - D11" - }, - "StemCellCharacterization_differentiation_caption": "iPSCs were differentiated to cardiomyocytes and observed for initiation of beating starting at day 6. At ~day 12, cells were fixed and stained with anti-cardiac Troponin T (BD Biosciences) and gating was based on an isotype control. Ranges observed across multiple experiments are shown for Troponin T and Day of beating initiation; number of experiments is shown in (). ", - "StemCellCharacterization_rnaseq_fig": "", - "StemCellCharacterization_rnaseq_fig_caption": "", - "Supplementary_acknowledgements": "/data/pdfs/aics_acknowledgements.pdf", - "Protocols_culturing": "/data/pdfs/AICS_WTC_culture SOP_external_v1-7_200210.pdf", - "Protocols_transfection": "/data/pdfs/AICS_SOP_RNP Transfection.pdf", - "Protocols_imaging": "/data/pdfs/AICS_SOP_WTC cell plating on glass for imaging.pdf", - "Protocols_ddpcr": "/data/pdfs/ddPCR_CNV_ASSAY_170526_Final.pdf", - "Protocols_cardio_diff": "/data/pdfs/sop_for_cardiomyocyte_differentiation_methods_v1.2_021020.pdf", - "Protocols_banking":"/data/pdfs/AICS_Banking_Guidelines_EXTERNAL_v1.3_200210.pdf", - "Stranalysis_report":"/data/pdfs/WTC11_STR_analysis.pdf" -}, + "StemCellCharacterization_differentiation": { + "troponin_percent_positive": "65.2", + "Day_of_beating_percent": "100 (4)", + "Day_of_beating_range": "D8 - D11" + }, + "StemCellCharacterization_differentiation_caption": "iPSCs were differentiated to cardiomyocytes and observed for initiation of beating starting at day 6. At ~day 12, cells were fixed and stained with anti-cardiac Troponin T (BD Biosciences) and gating was based on an isotype control. Ranges observed across multiple experiments are shown for Troponin T and Day of beating initiation; number of experiments is shown in (). ", + "StemCellCharacterization_rnaseq_fig": "", + "StemCellCharacterization_rnaseq_fig_caption": "", + "Supplementary_acknowledgements": "/data/pdfs/aics_acknowledgements.pdf", + "Protocols_culturing": "/data/pdfs/AICS_WTC_culture SOP_external_v1-7_200210.pdf", + "Protocols_transfection": "/data/pdfs/AICS_SOP_RNP Transfection.pdf", + "Protocols_imaging": "/data/pdfs/AICS_SOP_WTC cell plating on glass for imaging.pdf", + "Protocols_ddpcr": "/data/pdfs/ddPCR_CNV_ASSAY_170526_Final.pdf", + "Protocols_cardio_diff": "/data/pdfs/sop_for_cardiomyocyte_differentiation_methods_v1.2_021020.pdf", + "Protocols_banking": "/data/pdfs/AICS_Banking_Guidelines_EXTERNAL_v1.3_200210.pdf", + "Stranalysis_report": "/data/pdfs/WTC11_STR_analysis.pdf" + }, { "cell_line_id": "AICS-0126", "new": "true", From 28b31dc8e7a90fca3002f9203acc20a723e724a8 Mon Sep 17 00:00:00 2001 From: jazmingarcia-github Date: Fri, 31 May 2024 13:12:05 -0700 Subject: [PATCH 2/4] Updated Stem Cell Characteristics for AICS-0126 clones made the sub pages to be true --- data/cell_line_catalog.json | 8 ++------ 1 file changed, 2 insertions(+), 6 deletions(-) diff --git a/data/cell_line_catalog.json b/data/cell_line_catalog.json index 327855d..943c34a 100644 --- a/data/cell_line_catalog.json +++ b/data/cell_line_catalog.json @@ -12186,7 +12186,7 @@ }, { "id": "stemcellcharacteristics", - "done": "false" + "done": "true" }, { "id": "supplementaryinformation", @@ -12375,7 +12375,7 @@ }, { "id": "stemcellcharacteristics", - "done": "false" + "done": "true" }, { "id": "supplementaryinformation", @@ -12502,10 +12502,6 @@ { "Marker": "SSEA-4", "positiveCells": "100.0" - }, - { - "Marker": "TRA-160", - "positiveCells": "100.0" } ], "StemCellCharacterization_pluripotency_analysis_caption": "iPSCs were stained with directly conjugated antibodies from BD Biosciences, acquired using a FACSAria III Fusion (BD Biosciences), and analyzed using FlowJo software (Treestar, Inc.). Doublets were excluded, then marker-specific gates were set according to corresponding fluorescence-minus-one (FMO) controls.", From 40d49af1fe2c70fbe59bd784ff296f08f4095c86 Mon Sep 17 00:00:00 2001 From: jazmingarcia-github Date: Fri, 31 May 2024 13:25:13 -0700 Subject: [PATCH 3/4] Added Assay Caption for SCC AICS-0126 cl 68 --- data/cell_line_catalog.json | 234 ++++++++++++++++++------------------ 1 file changed, 117 insertions(+), 117 deletions(-) diff --git a/data/cell_line_catalog.json b/data/cell_line_catalog.json index 943c34a..b45418a 100644 --- a/data/cell_line_catalog.json +++ b/data/cell_line_catalog.json @@ -12355,64 +12355,65 @@ "Protocols_banking": "/data/pdfs/AICS_Banking_Guidelines_EXTERNAL_v1.3_200210.pdf", "Stranalysis_report": "/data/pdfs/WTC11_STR_analysis.pdf" }, -{ - "cell_line_id": "AICS-0126", - "new": "true", - "clone_number": "68", - "alleleCount": "mono", - "status": "Yes", - "subpaged_status": [ - { "id": "editingdesign", - "done": "true" - }, - { - "id":"genomiccharacterizations", - "done": "true" - }, - { - "id": "validation", - "done": "false" - }, - { - "id": "stemcellcharacteristics", - "done": "true" - }, - { - "id": "supplementaryinformation", - "done": "true" - }, - { - "id": "protocols", - "done": "true" - }, - { - "id": "stranalysis", - "done": "true" - } - ], - "Main_data_available": "true", - "Main_parent_line": "WTC", - "Main_gene_symbol": ["CDH5"], - "Main_gene_name": "cadherin 5", - "Main_protein": "Vascular endothelial (VE)-cadherin", - "Main_gene_id": [1003], - "Main_isoforms": [ - { - "gene_name": "CDH5", - "gene_isoforms": ["NM_001795.5","XM_047433470.1", "XM_047433469.1", "XM_047433471.1"] - } - ], - "Main_structure": "endothelial adherens junctions", - "Main_terminal_tagged": "C-terminus", - "Main_fluorescent_tag": "mEGFP", - "Main_linker": "N/A", - "Main_order_link": "https://www.coriell.org/0/Sections/Search/Sample_Detail.aspx?Ref=AICS-0126-068&Product=iPSC&PgId=166", - "Main_cofa": "https://www.coriell.org/0/PDF/Allen/ipsc/AICS-0126-068_CofA.pdf", - "Main_parental_order_link": "https://catalog.coriell.org/0/Sections/Search/Sample_Detail.aspx?Ref=GM25256&Product=CC", - "Main_donor_plasmid": "", - "Main_eu_hpsc_reg": "https://hpscreg.eu/cell-line/UCSFi001-A-66", + { + "cell_line_id": "AICS-0126", + "new": "true", + "clone_number": "68", + "alleleCount": "mono", + "status": "Yes", + "subpaged_status": [ + { + "id": "editingdesign", + "done": "true" + }, + { + "id": "genomiccharacterizations", + "done": "true" + }, + { + "id": "validation", + "done": "false" + }, + { + "id": "stemcellcharacteristics", + "done": "true" + }, + { + "id": "supplementaryinformation", + "done": "true" + }, + { + "id": "protocols", + "done": "true" + }, + { + "id": "stranalysis", + "done": "true" + } + ], + "Main_data_available": "true", + "Main_parent_line": "WTC", + "Main_gene_symbol": [ "CDH5" ], + "Main_gene_name": "cadherin 5", + "Main_protein": "Vascular endothelial (VE)-cadherin", + "Main_gene_id": [ 1003 ], + "Main_isoforms": [ + { + "gene_name": "CDH5", + "gene_isoforms": [ "NM_001795.5", "XM_047433470.1", "XM_047433469.1", "XM_047433471.1" ] + } + ], + "Main_structure": "endothelial adherens junctions", + "Main_terminal_tagged": "C-terminus", + "Main_fluorescent_tag": "mEGFP", + "Main_linker": "N/A", + "Main_order_link": "https://www.coriell.org/0/Sections/Search/Sample_Detail.aspx?Ref=AICS-0126-068&Product=iPSC&PgId=166", + "Main_cofa": "https://www.coriell.org/0/PDF/Allen/ipsc/AICS-0126-068_CofA.pdf", + "Main_parental_order_link": "https://catalog.coriell.org/0/Sections/Search/Sample_Detail.aspx?Ref=GM25256&Product=CC", + "Main_donor_plasmid": "", + "Main_eu_hpsc_reg": "https://hpscreg.eu/cell-line/UCSFi001-A-66", - "Main_media": [ + "Main_media": [ { "type": "image", "cell_line_id": "AICS-0126", @@ -12429,16 +12430,16 @@ "caption": "One vial of distribution lot was thawed (cells were treated with ROCK inhibitor for 24hrs post-thaw-refer to culture protocol). Cultures were observed daily. Colonies were imaged one and three days post-thaw using a Leica microscope at 4x and 10x magnification.", "link": "/images/AICS-0126/AICS-126_cl68_CDH5.jpg" } - ], - "EditingDesign_crRNA_seq": "GGAGGAGCTGCTGTATTAGG", - "EditingDesign_cas9": "Wildtype spCas9", - "EditingDesign_NCBI_isoform": "", - "EditingDesign_gene_figure": "/images/AICS-0126/CDH5_GFP.png", - "EditingDesign_gene_figure_caption": "Top: CDH5 locus showing 3 CDH5 isoforms; Bottom: Zoom in on mEGFP insertion site at CDH5 C-terminal exon.", - "GenomicCharacterization_junction_schematic": "/images/GenomicCharacterization_junction_schematic_mEGFP.png", - "GenomicCharacterization_tagged_allele_gel": "/images/AICS-0126/CDH5_cl68_full_allele.jpg", + ], + "EditingDesign_crRNA_seq": "GGAGGAGCTGCTGTATTAGG", + "EditingDesign_cas9": "Wildtype spCas9", + "EditingDesign_NCBI_isoform": "", + "EditingDesign_gene_figure": "/images/AICS-0126/CDH5_GFP.png", + "EditingDesign_gene_figure_caption": "Top: CDH5 locus showing 3 CDH5 isoforms; Bottom: Zoom in on mEGFP insertion site at CDH5 C-terminal exon.", + "GenomicCharacterization_junction_schematic": "/images/GenomicCharacterization_junction_schematic_mEGFP.png", + "GenomicCharacterization_tagged_allele_gel": "/images/AICS-0126/CDH5_cl68_full_allele.jpg", "GenomicCharacterization_tagged_allele_gel_caption": "Junctional PCR primers complementary to sequences flanking the homology arms in the distal genome were used together to co-amplify tagged and untagged alleles. Gel shows untagged allele (smaller band) and tagged allele (larger band) from single PCR reaction. (1% Agarose gel with Thermo Fisher 1kb ladder). Clone 68 is available from Coriell.", - "GenomicCharacterization_junction_table": [ + "GenomicCharacterization_junction_table": [ { "editedGene": "CDH5-mEGFP", "junction": "5'", @@ -12463,26 +12464,26 @@ "expected_size": "Edited allele = 2528, Unedited allele = 1811", "confirmed_sequence": "Yes" } - ], - "GenomicCharacterization_junction_table_caption": "PCR amplified 5', 3', WT, and full allele junctions. 5', 3', and WT junctions were Sanger sequenced to check for precise mEGFP insertion.", + ], + "GenomicCharacterization_junction_table_caption": "PCR amplified 5', 3', WT, and full allele junctions. 5', 3', and WT junctions were Sanger sequenced to check for precise mEGFP insertion.", - "GenomicCharacterization_ddpcr": [ + "GenomicCharacterization_ddpcr": [ { "gene_tag": "CDH5-mEGFP", "clone": "cl68", "fp_ratio": "0.577", "plasmid": "N/A" } - ], - "GenomicCharacterization_ddpcr_caption": "FP:RPP30 ratio from ddPCR assay; values = 0.5 +/- 0.1 indicate heterozygous clone, values = 1 +/- 0.1 indicate homozygous clone. RPP30 is known 2n reference gene.", - "GenomicCharacterization_offtargets": "", - "GenomicCharacterization_offtargets_caption": "", - "Validation_WesternBlot": "", - "Validation_WesternBlot_caption": "", - "Validation_Immunofluorescence": "", - "Validation_Immunofluorescence_caption": "", + ], + "GenomicCharacterization_ddpcr_caption": "FP:RPP30 ratio from ddPCR assay; values = 0.5 +/- 0.1 indicate heterozygous clone, values = 1 +/- 0.1 indicate homozygous clone. RPP30 is known 2n reference gene.", + "GenomicCharacterization_offtargets": "", + "GenomicCharacterization_offtargets_caption": "", + "Validation_WesternBlot": "", + "Validation_WesternBlot_caption": "", + "Validation_Immunofluorescence": "", + "Validation_Immunofluorescence_caption": "", - "StemCellCharacterization_pluripotency_analysis": [ + "StemCellCharacterization_pluripotency_analysis": [ { "Marker": "NANOG", "positiveCells": "98.6" @@ -12503,47 +12504,46 @@ "Marker": "SSEA-4", "positiveCells": "100.0" } - ], - "StemCellCharacterization_pluripotency_analysis_caption": "iPSCs were stained with directly conjugated antibodies from BD Biosciences, acquired using a FACSAria III Fusion (BD Biosciences), and analyzed using FlowJo software (Treestar, Inc.). Doublets were excluded, then marker-specific gates were set according to corresponding fluorescence-minus-one (FMO) controls.", - "StemCellCharacterization_karyotype": "/images/AICS-0126/AICS-126_cl68_CDH5_karyotype.jpg", - "StemCellCharacterization_karyotype_caption": "After cells banks were created, one vial was thawed and 30 G-banded metaphase cells were karyotyped.", - "StemCellCharacterization_trilineage": [ - { - "Germ_Layer": "Ectoderm", - "Marker": "PAX6", - "percent_positive_cells": 98.4 - }, - { - "Germ_Layer": "Endoderm", - "Marker": "SOX17", - "percent_positive_cells": 93.8 - }, - { - "Germ_Layer": "Mesoderm", - "Marker": "Brachyury", - "percent_positive_cells": 95.1 - } - ], - "StemCellCharacterization_trilineage_caption": "iPSCs were subjected to a 5-7 day, non-terminal, directed differentiation using the STEMdiff™ Trilineage Differentiation Kit (STEMCELL Technologies, Inc.). Total RNA was isolated from each lineage specific differentiation and assayed via ddPCR for the expression of lineage specific transcripts; Pax6(Ectoderm), Sox17(Endoderm) and Brachyury(Mesoderm).", - - "StemCellCharacterization_differentiation": + ], + "StemCellCharacterization_pluripotency_analysis_caption": "iPSCs were stained with directly conjugated antibodies from BD Biosciences, acquired using a CytoFLEX (Beckman Coulter), and analyzed using FlowJo software (Treestar, Inc.). Doublets were excluded, then marker-specific gates were set according to corresponding fluorescence-minus-one (FMO) controls.", + "StemCellCharacterization_karyotype": "/images/AICS-0126/AICS-126_cl68_CDH5_karyotype.jpg", + "StemCellCharacterization_karyotype_caption": "After cells banks were created, one vial was thawed and 30 G-banded metaphase cells were karyotyped.", + "StemCellCharacterization_trilineage": [ { - "troponin_percent_positive": "65.2", - "Day_of_beating_percent": "100 (4)", - "Day_of_beating_range": "D8 - D11" + "Germ_Layer": "Ectoderm", + "Marker": "PAX6", + "percent_positive_cells": 98.4 }, - "StemCellCharacterization_differentiation_caption": "iPSCs were differentiated to cardiomyocytes and observed for initiation of beating starting at day 6. At ~day 12, cells were fixed and stained with anti-cardiac Troponin T (BD Biosciences) and gating was based on an isotype control. Ranges observed across multiple experiments are shown for Troponin T and Day of beating initiation; number of experiments is shown in (). ", - "StemCellCharacterization_rnaseq_fig": "", - "StemCellCharacterization_rnaseq_fig_caption": "", - "Supplementary_acknowledgements": "/data/pdfs/aics_acknowledgements.pdf", - "Protocols_culturing": "/data/pdfs/AICS_WTC_culture SOP_external_v1-7_200210.pdf", - "Protocols_transfection": "/data/pdfs/AICS_SOP_RNP Transfection.pdf", - "Protocols_imaging": "/data/pdfs/AICS_SOP_WTC cell plating on glass for imaging.pdf", - "Protocols_ddpcr": "/data/pdfs/ddPCR_CNV_ASSAY_170526_Final.pdf", - "Protocols_cardio_diff": "/data/pdfs/sop_for_cardiomyocyte_differentiation_methods_v1.2_021020.pdf", - "Protocols_banking":"/data/pdfs/AICS_Banking_Guidelines_EXTERNAL_v1.3_200210.pdf", - "Stranalysis_report":"/data/pdfs/WTC11_STR_analysis.pdf" -}, + { + "Germ_Layer": "Endoderm", + "Marker": "SOX17", + "percent_positive_cells": 93.8 + }, + { + "Germ_Layer": "Mesoderm", + "Marker": "Brachyury", + "percent_positive_cells": 95.1 + } + ], + "StemCellCharacterization_trilineage_caption": "iPSCs were subjected to a 5-7 day, non-terminal, directed differentiation using the STEMdiff™ Trilineage Differentiation Kit (STEMCELL Technologies, Inc.). Total RNA was isolated from each lineage specific differentiation and assayed via ddPCR for the expression of lineage specific transcripts; Pax6(Ectoderm), Sox17(Endoderm) and Brachyury(Mesoderm).", + + "StemCellCharacterization_differentiation": { + "troponin_percent_positive": "65.2", + "Day_of_beating_percent": "100 (4)", + "Day_of_beating_range": "D8 - D11" + }, + "StemCellCharacterization_differentiation_caption": "iPSCs were differentiated to cardiomyocytes and observed for initiation of beating starting at day 6. At ~day 12, cells were fixed and stained with anti-cardiac Troponin T (BD Biosciences) and gating was based on an isotype control. Ranges observed across multiple experiments are shown for Troponin T and Day of beating initiation; number of experiments is shown in (). ", + "StemCellCharacterization_rnaseq_fig": "", + "StemCellCharacterization_rnaseq_fig_caption": "", + "Supplementary_acknowledgements": "/data/pdfs/aics_acknowledgements.pdf", + "Protocols_culturing": "/data/pdfs/AICS_WTC_culture SOP_external_v1-7_200210.pdf", + "Protocols_transfection": "/data/pdfs/AICS_SOP_RNP Transfection.pdf", + "Protocols_imaging": "/data/pdfs/AICS_SOP_WTC cell plating on glass for imaging.pdf", + "Protocols_ddpcr": "/data/pdfs/ddPCR_CNV_ASSAY_170526_Final.pdf", + "Protocols_cardio_diff": "/data/pdfs/sop_for_cardiomyocyte_differentiation_methods_v1.2_021020.pdf", + "Protocols_banking": "/data/pdfs/AICS_Banking_Guidelines_EXTERNAL_v1.3_200210.pdf", + "Stranalysis_report": "/data/pdfs/WTC11_STR_analysis.pdf" + }, { "cell_line_id": "AICS-0122", "new": "", From 6697a8fbb5273727dd9de9a09a7380a39c3f6299 Mon Sep 17 00:00:00 2001 From: jazmingarcia-github Date: Wed, 5 Jun 2024 10:05:59 -0700 Subject: [PATCH 4/4] updating western blot for AISC-0126 mono and bi clones --- data/cell_line_catalog.json | 8 ++++---- ...ReleaseWestern_AICS0126_CDH5_cl41_bi_V3.jpg | Bin 0 -> 119374 bytes ...leaseWestern_AICS0126_CDH5_cl68_mono_V3.jpg | Bin 0 -> 121212 bytes 3 files changed, 4 insertions(+), 4 deletions(-) create mode 100644 images/AICS-0126/ReleaseWestern_AICS0126_CDH5_cl41_bi_V3.jpg create mode 100644 images/AICS-0126/ReleaseWestern_AICS0126_CDH5_cl68_mono_V3.jpg diff --git a/data/cell_line_catalog.json b/data/cell_line_catalog.json index b45418a..308863d 100644 --- a/data/cell_line_catalog.json +++ b/data/cell_line_catalog.json @@ -12182,7 +12182,7 @@ }, { "id": "validation", - "done": "false" + "done": "true" }, { "id": "stemcellcharacteristics", @@ -12289,7 +12289,7 @@ "GenomicCharacterization_ddpcr_caption": "FP:RPP30 ratio from ddPCR assay; values = 0.5 +/- 0.1 indicate heterozygous clone, values = 1 +/- 0.1 indicate homozygous clone. RPP30 is known 2n reference gene.", "GenomicCharacterization_offtargets": "", "GenomicCharacterization_offtargets_caption": "", - "Validation_WesternBlot": "", + "Validation_WesternBlot": "/images/AICS-0126/ReleaseWestern_AICS0126_CDH5_cl41_bi_V3.jpg", "Validation_WesternBlot_caption": "", "Validation_Immunofluorescence": "", "Validation_Immunofluorescence_caption": "", @@ -12372,7 +12372,7 @@ }, { "id": "validation", - "done": "false" + "done": "true" }, { "id": "stemcellcharacteristics", @@ -12478,7 +12478,7 @@ "GenomicCharacterization_ddpcr_caption": "FP:RPP30 ratio from ddPCR assay; values = 0.5 +/- 0.1 indicate heterozygous clone, values = 1 +/- 0.1 indicate homozygous clone. 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