STEMCELL Technologies mTeSR iPSCdirect Healthy Control Human iPSC Line
- 研究用
- 新製品
iPSCdirect™ Healthy Control Human iPSC Line (ST-200-0510) は、すぐに使えるシングルセル仕様のヒト人工多能性幹細胞 (iPSC) SCTi003-A 株です。一貫した高品質で使い切りの iPSCdirect™ 細胞を使用すれば、iPSC研究用セルバンクの開発や特性解析にかかるコストと労力を減らして長期培養を回避でき、研究のスピードアップに役立ちます。本品は、解凍後ただちにiPSCベースのスクリーニングや様々な細胞種への分化に使用可能です。
本品は、iPSCコントロールラインである SCTi003-A から特別に凍結保存されています。製品性能を最適化するため、mTeSR™ Plus 培地を用いて製造され、単層ベースの分化プロトコール用に最適化されています。本品は、STEMdiff™ Ventricular Cardiomyocyte Differentiation Kit、STEMdiff™ SMADi Neural Induction Kit、STEMdiff™ Hepatocyte Kit などの STEMdiff™ 培地製品を用いた分化に適しています。
本品は研究用（RUO）で、学術および商用の両方での使用が許諾されています。ソースセルバンクのドナーの詳細と細胞品質の特徴は、データ紹介欄をご参照ください。その他の詳細については、ロット固有の検査証明書（CoA）をご参照ください。
2018/05/14 12:00の製品情報
本製品は研究目的にのみ使用し、人や動物の医療用・臨床診断用・食品用としては使用しないようにご注意ください。
製品の特長
iPSCdirect™は、シングルユースでメンテナンス不要のヒトiPS細胞凍結品です
- 播種後24時間の単層培養で分化準備が整い、iPS細胞の維持が不要になります
- 親細胞株の SCTi003-A と同様、業界標準以上の広範な品質管理を満たしています
- すぐに使用できて高密度、かつ多くの分化用キット (STEMdiff™) * に適合しており、研究を加速出来ます
* 分化のデータは、こちら＞＞
iPSCdirect™ 単層培養の流れ
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.
データ紹介
播種密度を調整することで、24時間後にさまざまなコンフルエンシーに到達します
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.
SCTi003-A 細胞株は健康な女性ドナー由来です
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 based on 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.
典型的な核型を維持しています
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.
コピー数多型 (CNV) は親細胞株と一致しています
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 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.
未分化細胞マーカーを発現しています
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.
3胚葉への高い分化能を示します
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.
