iPSCdirect™ Healthy Control Human iPSC Line SCTi003-A
Human induced pluripotent stem cells, frozen
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mTeSR™ PluscGMP, stabilized feeder-free maintenance medium for human ES and iPS cells
Overview
iPSCdirect™ cells are specially cryopreserved from the iPSC control line, SCTi003-A, which is derived from healthy female donor peripheral blood mononuclear cells (PBMCs). For optimal product performance and reproducibility, iPSCdirect™ is manufactured and maintained in mTeSR™ Plus, is characterized using extensive quality control procedures, and has been performance-tested with a number of STEMdiff™ kits. Browse TeSR™ and STEMdiff™ media products to establish a complete workflow for your cell culture system.
This research-use-only (RUO) product has been consented for both academic and commercial use. Blood samples are ethically sourced using Institutional Review Board (IRB) or other regulatory authority-approved consent forms and protocols. For donor details and cell quality characterization of the source cell banks, refer to the data figures on this page. Registration with hPSCreg® ensures ethical and biological conformity based on community standards. For additional details, refer to the lot-specific Certificate of Analysis and Frequently Asked Questions about iPSC lines.
Data Figures
Figure 1. Schematic for a Generalized Monolayer Protocol to Thaw and Culture iPSCdirect™ for iPSC-Based Monolayer Workflows
iPSCdirect™ cells can be thawed and plated into mTeSR™ Plus (Catalog #100-0276) supplemented with CloneR™2 (Catalog #100-0691) and incubated overnight according to product instructions. Recommendations for seeding densities to reach the desired confluency at 24 hours may be found in the Product Information Sheet. After 24 hours, cells are ready for STEMdiff™ or customized monolayer workflows. iPSC = induced pluripotent stem cell.
Figure 2. iPSCdirect™ SCTi003-A Cells Can Be Seeded to Reach a Range of Confluencies After 24 Hours
To reach the desired confluency for downstream experiments, thaw and plate iPSCdirect™ cells into mTeSR™ Plus with CloneR™2 (Catalog #100-0276 and #100-0691) at the densities recommended in the Product Information Sheet. These representative examples of (A) low confluency, (B) medium confluency, and (C) high confluency were cultured after thaw on Corning® Matrigel® hESC-Qualified Matrix and imaged at a magnification of 4X.
Figure 3. iPSC Line SCTi003-A, the Source for iPSCdirect™ SCTi003-A, Is Derived from a Healthy Female Donor
Demographic, health, and genetic characteristics of the SCTi003-A donor are compiled from self-reported information and whole-exome sequencing. Sex was determined by karyotype. Ancestry was calculated by EthSEQ analysis from whole-exome sequencing data. HLA haplotype was determined by next-generation sequencing, sequence-base typing, and sequence-specific oligonucleotide probes as needed to obtain the required resolution. Other genetic variants were determined from whole-exome sequencing using ClinVar analysis. Blood type (ABO/Rh blood group) was determined by next-generation sequencing. Height, weight, and BMI were calculated at the donation facility.
Figure 4. iPSCdirect™ SCTi003-A Human Pluripotent Stem Cells Maintain a Typical Karyotype
G-T-L banding for thawed iPSCdirect™ cells (SCTi003-A p34, n = 40) shows a typical karyotype with no evidence of clonal abnormalities at a band resolution of 425 - 475 G-bands per haploid genome.
Figure 5. Single Nucleotide Polymorphism Microarray Analysis Characterizes iPSCdirect™ SCTi003-A Copy Number Variants Consistent with the Source Cell Line
DNA was extracted from a vial of iPSCdirect™ SCTi003-A iPSCs and subjected to SNP microarray analysis to identify large-scale copy number variants (CNVs). Consistent with the variants detected in the source SCTi003-A cell line (see Figure 4 of SCTi003-A product data), the thawed cells display two reportable CNVs, defined as those greater than 400kb in size, on chromosome 7 and 14 (rows highlighted in bold font). These losses are located in the TCR regions of the genome and are indicative of VDJ recombination process during T Cell development. Array design, genomics position, genes, and chromosome banding are based on genome build GRCh37/hg19. chr = chromosome; start cyto = cytogenetic band at the start of the base pair imbalance; end cyto = cytogenetic band at the end of the base pair imbalance; bp = base pairs; SNP = single nucleotide polymorphism; TCR = T Cell Receptor; VDJ = variable, diversity, joining segment.
Figure 6. iPSCdirect™ SCTi003-A Cells Express Undifferentiated Cell Markers
iPSCdirect™ SCTi003-A was characterized using flow cytometry for undifferentiated cell markers OCT3/4 and TRA-1-60. Percentage marker expression was quantified 4 days after thawing, from analyses of three biological replicates (n = 3 vials). Error bars represent the standard deviation.
Figure 7. iPSCdirect™ SCTi003-A Human Pluripotent Stem Cells Demonstrate a High Trilineage Differentiation Capacity
iPSCdirect™ SCTi003-A cells were split into 3 groups, differentiated using STEMdiff™ Trilineage Differentiation Kit (Catalog #05230), and then subjected to flow cytometry analysis. Two markers for each embryonic germ layer were assessed, and bars represent mean marker expression for each group of cells (dots represent the average of 3 technical replicates; error bars represent standard deviation; n = 3 biological replicates). All lineage-specific markers were expressed by more than 70% of differentiated cells. Expression of PAX6 and Nestin confirm differentiation to the ectoderm lineage, NCAM and Brachyury (T) expression confirm differentiation to the mesoderm lineage, and CXCR4 and SOX17 expression confirm differentiation to the endoderm lineage.
Figure 8. iPSCdirect™ SCTi003-A Human Pluripotent Stem Cells Can Efficiently Differentiate into Neural Progenitor Cells Upon Thawing
NPCs were generated from iPSCdirect™ SCTi003-A cells using the protocol in the iPSCdirect™ Product Information Sheet followed by the monolayer protocol from Day 1 in the STEMdiff™ SMADi Neural Induction Kit Technical Manual (Catalog #08581). After the first 7 days of the protocol, the resulting NPCs were fixed for immunocytochemistry. The NPCs expressed neural progenitor markers (A) SOX1 and (B) PAX6, and nuclei were visualized with (C) DAPI. (D) Marker expression was quantified and negative control SOX10 expression was minimal. Error bars represent standard deviation (n = 2 biological replicates). NPCs = neural progenitor cells.
Figure 9. iPSCdirect™ SCTi003-A Human Pluripotent Stem Cells Can Successfully Differentiate into Ventricular Cardiomyocytes
Ventricular cardiomyocytes were generated from iPSCdirect™ SCTi003-A cells using STEMdiff™ Ventricular Cardiomyocyte Differentiation Kit (Catalog #05010). (A) 48 hours after thawing and plating in mTeSR™ Plus and CloneR™2, iPSCdirect™ cells reached the desired confluency and are ready for Day 0 of differentiation according to the STEMdiff™ Ventricular Cardiomyocyte Product Information Sheet. (B) By Day 15 of differentiation, monolayer cultures show iPSC-derived ventricular cardiomyocytes that (C) exhibit coordinated beating behavior.
Figure 10. iPSCdirect™ SCTi003-A Human Pluripotent Stem Cells Can Successfully Differentiate into Hepatocytes
iPSCdirect™ SCTi003-A cells were plated according to the iPSCdirect™ Product Information Sheet, in preparation for Day 1 of the STEMdiff™ Hepatocyte Kit (Catalog #100-0520) protocol outlined in the STEMdiff™ Hepatocyte Kit Product Information Sheet. (A) Monolayers of hepatocyte-like cells with characteristic polygonal morphology and binucleation are apparent by Day 21 of differentiation. (B) Marker expression was quantified by flow cytometry for hepatic markers albumin (ALB) and alpha1-antitrypsin (A1AT). Error bars represent standard deviation (n = 3 biological replicates).
Protocols and Documentation
Find supporting information and directions for use in the Product Information Sheet or explore additional protocols below.
Applications
This product is designed for use in the following research area(s) as part of the highlighted workflow stage(s). Explore these workflows to learn more about the other products we offer to support each research area.
Resources and Publications
Educational Materials (5)
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Legal Statement:
These iPSCs and their modifications (including but not limited to derivatives or differentiated progeny) shall not be used or administered in (1) human subjects for human clinical use; (2) animals for veterinary use for therapeutic, diagnostic, or prophylactic purposes or (3) any subject in relation to clinical applications, cell therapy, transplantation, and/or regenerative medicines, without limiting the generality of the foregoing. These iPSCs and their modifications (including but not limited to derivatives or differentiated progeny) may not be used for monetization or commercialization purposes, including without limitation, used to, or with the goal to, perform services or supply products or rights, including in the manufacture of cellular therapies or other therapeutics, for monetary gain or the generation of royalties, revenues, sales or other valuable consideration. For clarity, these iPSCs and their modifications (including but not limited to derivatives or differentiated progeny) may not be used for screening compounds, antibodies, proteins or peptides, except for the purposes of target discovery, target validation, or assay development, provided such activities and the results of such activities are not further used for monetization or commercialization purposes. It may be possible to obtain a further license for the prohibited uses referred to in this Limited Use License. Please contact iPSCrequests@stemcell.com for more details. PRODUCTS ARE FOR RESEARCH USE ONLY AND NOT INTENDED FOR HUMAN OR ANIMAL DIAGNOSTIC OR THERAPEUTIC USES UNLESS OTHERWISE STATED. Copyright © 2023 by STEMCELL Technologies Inc. All rights reserved including graphics and images. STEMCELL Technologies & Design, STEMCELL Shield Design, Scientists Helping Scientists, CloneR, ReLeSR, and STEMdiff are trademarks of STEMCELL Technologies Canada Inc. mTeSR and TeSR are trademarks of WARF. Corning, Falcon, and Matrigel are registered trademarks of Corning Incorporated. CryoStor and ThawStar are registered trademarks of BioLife Solutions. All other trademarks are the property of their respective holders. While STEMCELL has made all reasonable efforts to ensure that the information provided by STEMCELL and its suppliers is correct, it makes no warranties or representations as to the accuracy or completeness of such information.
Quality Statement:
PRODUCTS ARE FOR RESEARCH USE ONLY AND NOT INTENDED FOR HUMAN OR ANIMAL DIAGNOSTIC OR THERAPEUTIC USES UNLESS OTHERWISE STATED. FOR ADDITIONAL INFORMATION ON QUALITY AT STEMCELL, REFER TO WWW.STEMCELL.COM/COMPLIANCE.
