iPSCdirect™

一次性使用、免维持的人诱导多能干细胞，冷冻

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产品号 #（选择产品）

产品号 #200-0510_C

一次性使用、免维持的人诱导多能干细胞，冷冻

产品优势

单层培养物在接种 24 小时后即可分化，无需对 hPSC 进行维持
预计获得与亲本细胞系 SCTi003-A 相同的、满足或超过行业标准的严格质量控制
高密度即用型 iPSC 兼容多种 STEMdiff™ 分化试剂盒，加速您的研究

产品组分包括

iPSCdirect™健康对照人iPSCs， SCTi003-A细胞系（产品号#200-0510）

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要查看实验方案所需的所有配套产品，请参阅《实验方案与技术文档》。

mTeSR™ Plus
cGMP, stabilized feeder-free maintenance medium for human ES and iPS cells
Labeling Antibodies
Compatible antibodies for purity assessment of isolated cells

总览

实验数据

产品说明书及文档

应用领域

总览

使用即用型单细胞 iPSCdirect™ 人诱导多能干细胞 (iPSC) 加速您的研究。iPSCdirect™ 细胞具有高度一致性和稳定性，可让您免去为 iPSC 研究开发和鉴定细胞库的成本和精力，也无需长期培养 iPSC。每次实验均使用相同来源且经过质量控制测试的 iPSC，可节省时间并减少差异性。iPSCdirect™ 可在复苏后直接用于基于 iPSC 的筛选或分化为多种细胞类型。

iPSCdirect™ 来源于健康对照细胞系SCTi003-A，采用专门冻存工艺保存。为了获得最佳产品性能，iPSCdirect™ 采用我领先的mTeSR™Plus培养基生产，并针对单层分化方案进行了优化。它与多种STEMdiff™培养基产品兼容，包括STEMdiff™心室肌细胞分化试剂盒, STEMdiff™SMADi神经诱导试剂盒,STEMdiff™肝细胞试剂盒.

本产品仅供研究使用（RUO），已获得学术和商业用途许可。有关供体详情和来源细胞库的细胞质量表征，请参阅本页的数据图表。如需更多细节，更多详细信息，请参阅特定批次的分析证书。

包含
FreSR™-S

亚型
冻存

细胞类型
多能干细胞

种属
人

细胞和组织来源
多能干细胞

品牌
STEMdiff，TeSR

研究领域
疾病建模，药物发现和毒理检测，感染性疾病(传染病)，神经科学

供体状态
Normal

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实验数据

Experimental protocol diagram for using iPSCdirect ready-to-use cells

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.

Microscopy images of iPSCdirect cells at low, medium, and high confluency after 24 hours

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.

Table 1. iPSC Line 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 and HLA haplotype were calculated from whole-genome and whole-exome sequencing combined data. Blood type (ABO/Rh blood group) was determined by next-generation sequencing. Height, weight, and BMI were calculated at the donation facility. iPSC = induced pluripotent stem cell.

Typical human karyogram showing all chromosomes and no clonal abnormalities

Figure 3. 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.

Table 2. Single Nucleotide Polymorphism Microarray Analysis Characterizes SCTi003-A Copy Number Variants

Table displaying single nucleotide polymorphism microarray analysis of the hiPSC control line SCTi003-A.

DNA was extracted from a vial of SCTi003-A from the Master Cell Bank and subject to SNP microarray analysis to identify large-scale copy number variants (CNVs). The 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.

Bar graph quantification of OCT3/4 and TRA-1-60 gene expression

Figure 4. 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.

Bar graph quantifying gene expression of 6 trilineage markers by flow cytometry

Figure 5. 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.

Immunofluorescent images of 2 neural progenitor markers and a nuclear stain along with their quantification

Figure 6. 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.

Microscopy images of iPSCdirect cells and differentiated ventral cardiomyocytes, and a video of coordinated contraction or beating behavior of cardiomyocytes in a culture dish

Figure 7. 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.

Microscopy image of Day 21 hepatocyte-like cells and quantification of hepatic gene expression markers ALB and A1AT

Figure 8. 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).