cryopreserved primary human hepatocytes (Thermo Fisher)

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Thermo Fisher cryopreserved primary human hepatocytes

Validation of cAAV in primary human <t>hepatocytes</t> (A) Schematic of cAAV design and mechanism. Orthogonal attachment sites (attB∗, attP∗) are added to an attP AAV genome to facilitate Bxb1-mediated intramolecular recombination. The reaction produces a circular dsDNA attP cargo and an ITR by-product. (B) Design of a ssAAV dual luciferase circularization reporter to validate cAAV function. In linear native format, EF1A promoter expresses Firefly luciferase, and NanoLuc is not expressed. When circularized, EF1A promoter expresses NanoLuc in the circular episome and Firefly luciferase is not expressed in the ITR by-product. (C) Relative amounts of linear or circularized cAAV reporter in PHH on days 1 and 5 following Bxb1 mRNA transfection. (D) Absolute quantification of cAAV circularization frequency on day 5 following Bxb1 mRNA transfection in PHH. (E) I-PGI outcome with cAAV or scAAV cargo over a 6-day time course in PHH. Statistical significance was calculated using Student’s unpaired two tailed t test (ns indicates not significant, ∗ p < 0.05, ∗∗ p < 0.01). (F) The effect of staggering cargo circularization and I-PGI on seamless insertion junction outcome in PHH. Predose is Bxb1 mRNA transfected twice, on day 2 and on day 5. For E-F, I-PGI was performed at the PAH locus through a single transfection of synthetic atgRNA, nCas9-RT and Bxb1 mRNAs. For (D–F), data were collected using ddPCR, and values were calculated as described in . Data reflect the mean and standard deviation of three biological replicates.

Cryopreserved Primary Human Hepatocytes, supplied by Thermo Fisher, used in various techniques. Bioz Stars score: 90/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/cryopreserved+primary+human+hepatocytes/cryopreserved+primary+human+hepatocytes/pmc12166831-208-2-4
Average 90 stars, based on 1 article reviews

cryopreserved primary human hepatocytes - by Bioz Stars, 2026-09

90/100 stars

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1) Product Images from "Development of circular AAV cargos for targeted seamless insertion with large serine integrases"

Article Title: Development of circular AAV cargos for targeted seamless insertion with large serine integrases

Journal: Molecular Therapy. Methods & Clinical Development

doi: 10.1016/j.omtm.2025.101490

Validation of cAAV in primary human hepatocytes (A) Schematic of cAAV design and mechanism. Orthogonal attachment sites (attB∗, attP∗) are added to an attP AAV genome to facilitate Bxb1-mediated intramolecular recombination. The reaction produces a circular dsDNA attP cargo and an ITR by-product. (B) Design of a ssAAV dual luciferase circularization reporter to validate cAAV function. In linear native format, EF1A promoter expresses Firefly luciferase, and NanoLuc is not expressed. When circularized, EF1A promoter expresses NanoLuc in the circular episome and Firefly luciferase is not expressed in the ITR by-product. (C) Relative amounts of linear or circularized cAAV reporter in PHH on days 1 and 5 following Bxb1 mRNA transfection. (D) Absolute quantification of cAAV circularization frequency on day 5 following Bxb1 mRNA transfection in PHH. (E) I-PGI outcome with cAAV or scAAV cargo over a 6-day time course in PHH. Statistical significance was calculated using Student’s unpaired two tailed t test (ns indicates not significant, ∗ p < 0.05, ∗∗ p < 0.01). (F) The effect of staggering cargo circularization and I-PGI on seamless insertion junction outcome in PHH. Predose is Bxb1 mRNA transfected twice, on day 2 and on day 5. For E-F, I-PGI was performed at the PAH locus through a single transfection of synthetic atgRNA, nCas9-RT and Bxb1 mRNAs. For (D–F), data were collected using ddPCR, and values were calculated as described in . Data reflect the mean and standard deviation of three biological replicates.

Figure Legend Snippet: Validation of cAAV in primary human hepatocytes (A) Schematic of cAAV design and mechanism. Orthogonal attachment sites (attB∗, attP∗) are added to an attP AAV genome to facilitate Bxb1-mediated intramolecular recombination. The reaction produces a circular dsDNA attP cargo and an ITR by-product. (B) Design of a ssAAV dual luciferase circularization reporter to validate cAAV function. In linear native format, EF1A promoter expresses Firefly luciferase, and NanoLuc is not expressed. When circularized, EF1A promoter expresses NanoLuc in the circular episome and Firefly luciferase is not expressed in the ITR by-product. (C) Relative amounts of linear or circularized cAAV reporter in PHH on days 1 and 5 following Bxb1 mRNA transfection. (D) Absolute quantification of cAAV circularization frequency on day 5 following Bxb1 mRNA transfection in PHH. (E) I-PGI outcome with cAAV or scAAV cargo over a 6-day time course in PHH. Statistical significance was calculated using Student’s unpaired two tailed t test (ns indicates not significant, ∗ p < 0.05, ∗∗ p < 0.01). (F) The effect of staggering cargo circularization and I-PGI on seamless insertion junction outcome in PHH. Predose is Bxb1 mRNA transfected twice, on day 2 and on day 5. For E-F, I-PGI was performed at the PAH locus through a single transfection of synthetic atgRNA, nCas9-RT and Bxb1 mRNAs. For (D–F), data were collected using ddPCR, and values were calculated as described in . Data reflect the mean and standard deviation of three biological replicates.

Techniques Used: Biomarker Discovery, Luciferase, Transfection, Quantitative Proteomics, Two Tailed Test, Standard Deviation

Cell Culture:

Article Title: Acrylic-based culture plate format perfusion device to establish liver endothelial-epithelial interface.
Article Snippet: Microphysiological Systems (MPSs) or organs-on-chips, are microfluidic devices used to model human physiology in vitro.. Polydimethylsiloxane (PDMS) is the most widely used material for organs-on-chips due to its established fabrication methods and biocompatibility properties.. However, non-specific binding of small molecules limits PDMS for drug screening applications.

Chromatography:

Article Title: Quantitative chemometric phenotyping of three-dimensional liver organoids by Raman spectral imaging.
Article Snippet: Neutral lipids were stained using HCS LipidTOXTMRed Neutral Lipids (excitation/emission maxima 577/609 nm, Thermo Fisher Scientific, catalogue no. H34476) and visualized using Andor Dragonfly Spinning Disk Confocal microscope. .. Proteomic liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis Pelleted iHLC organoids generated from 3 cell lines (iHLC_1: WTC-11, WiCell; iHLC_2: WTSIi013-A and iHLC_3: WTSIi028-A, Wellcome Trust Sanger Institute) and cryopreserved primary human hepatocytes (Gibco, lot HU8287 (PHH_3)) (all 100 000-300 000 cells) were washed oncewith DPBS (final volume 15 mL), before lysing and digestion. .. Sample preparation was performed according to the protocol for sample preparation by Easy Extraction and Digestion (SPEED) by Doellinger et al.,15 with a modified reduction and alkylation step: reduction was performed by adding DL-dithiothreitol (Merck, catalogue nr. D5545) to a final concentration of 10 mM before incubation at 56 C / 900 rpm for 25 min in a thermoshaker (Grant instruments), and alkylation was performed by adding iodoacetamide (Merck, catalogue no. I1149) to a total concentration of 20 mM before incubation at room temperature / 900 rpm for 30 min in the thermoshaker (in the dark).

Mass Spectrometry:

Generated:

Recombinant:

Article Title: Quantitative chemometric phenotyping of three-dimensional liver organoids by Raman spectral imaging.
Article Snippet: .. REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Rabbit Human Serum Albumin Polyclonal antibody Abcam Cat# ab2406; RRID: AB_303048 Goat ZO-1 Polyclonal Antibody Thermo Fisher Scientific Cat# PA5-19090; RRID: AB_10988668 Mouse CYP3A4 Monoclonal Antibody (Clone 3H8) Thermo Fisher Scientific Cat# MA5-17064; RRID: AB_2538535 Mouse Anti-MRP2 Monoclonal Antibody (Clone M2 III-6) Abcam Cat# ab3373; RRID: AB_303751 Alexa Fluor 488-AffiniPure Donkey Anti-Goat IgG (H+L) (min X Ck,GP,Sy Hms,Hrs,Hu,Ms,Rb,Rat Sr Prot) Jackson ImmunoResearch Labs Cat# 705-545-147; RRID: AB_2336933 Cy3-AffiniPure Donkey Anti-Rabbit IgG (H+L) (min X Bov,Ck,Gt,GP,Sy Hms,Hrs,Hu,Ms,Rat,Shp Sr Prot) Jackson ImmunoResearch Labs Cat# 711-165-152; RRID: AB_2307443 Alexa Fluor 647 AffiniPure Donkey Anti-Mouse IgG (H+L) Jackson ImmunoResearch Labs Cat# 715-605-150; RRID: AB_2340862 Biological samples Cryopreserved primary human hepatocytes Gibco Cat# HMCPSQ; lot HU8339-A Cryopreserved primary human hepatocytes Lonza Cat# HUCPG; lot HUM180201A Cryopreserved primary human hepatocytes Gibco Cat# HMCPMS; lot HU8287 XTreme 200 POOL HUMAN LIVER MICROSOMES Tebubio Cat#H2630; Lot nr: 171008 Chemicals, peptides, and recombinant proteins Neratinib Selleckchem Cat# S2150 Nilotinib Selleckchem Cat# S1033 Ketoconazole Selleckchem Cat# S1353 Amiodarone HCl Selleckchem Cat# S1979 Fluticasone propionate Selleckchem Cat# S1992 Methadone stock solution This paper n/a Critical commercial assays CellTiter-Glo 3D Cell Viability Assay Promega Cat# G9681 P450-GloTM Assay with Luciferin-IPA Promega Cat# V9001 P450-GloTM CYP1A2 Assay Promega Cat# V8771 HCS LipidTOXTM Red neutral lipid stain Thermo Fisher Scientific Cat# H34476 Human Albumin ELISA Quantitation Set Bethyl Laboratories Cat# E88-129 Experimental models: Cell lines Induced pluripotent stem cell WTC-11 Coriell Institute for Medical Research RRID:CVCL_Y803 Induced pluripotent stem cell WTSIi013-A Wellcome Trust Sanger Institute RRID:CVCL_AE60 Induced pluripotent stem cell WTSIi028-A Wellcome Trust Sanger Institute RRID:CVCL_AI14 Software and algorithms Fiji Schneider et al.7 https://imagej.nih.gov/ij/ PEAKS X+ software version 10.5 Bioinformatics Solutions https://www.bioinfor.com/peaks-studio/ GraphPad PRISM 7 GraphPad Software Inc. All original code has been deposited at GitHub repository and is publicly available via Zenodo: https://doi.org/10.5281/zenodo.7628912 Developed in-house during the project https://doi.org/10.5281/zenodo.7628912 ..

Viability Assay:

Staining:

Enzyme-linked Immunosorbent Assay:

Quantitation Assay:

Software:

other:

Article Title: Induction of MASH in three-dimensional bioprinted human liver tissue
Article Snippet: 3D liver tissues comprising of cryopreserved primary human hepatocytes (Gibco, Thermo Fisher Scientific), hepatic stellate cells, liver endothelial cells, and Kupffer cells, were fabricated by Organovo, Inc. (San Diego, CA, USA) with a NovoGen Bioprinter onto 24-well transwell culture inserts using patented protocols as previously described [ – ].

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