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红系祖细胞重编程试剂盒

用于从外周血中分选和扩增红系祖细胞,并将其进一步重编程为诱导多能干细胞(iPS细胞)试剂盒

产品号 #(选择产品)

产品号 #05924_C

用于从外周血中分选和扩增红系祖细胞,并将其进一步重编程为诱导多能干细胞(iPS细胞)试剂盒

产品优势

  • 优化的富集,扩增和重编程红细胞祖细胞扩增外周血样本
  • 与传统的hES细胞培养基相比,提高了iPS细胞的重编程效率和集落频率
  • 具有高质量iPS细胞样形态的大菌落迅速出现,有利于鉴定和亚克隆
  • 与TeSR™和STEMdiff™产品无缝集成,用于iPS细胞系的下游维持和分化

产品组分包括

  • 针对来自外周血样本的红系祖细胞的富集、扩增及重编程过程进行了优化
  • 与传统人胚胎干细胞(hES)培养基相比,重编程效率更高,iPS细胞集落生成频率更高
  • 具有高质量 iPS 细胞样形态的大型克隆的快速出现有利于识别和亚克隆
  • 与 TeSR™ 和 STEMdiff™ 产品无缝衔接,可用于后续 iPS 细胞系的维持培养与分化应用
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总览

红细胞祖细胞重编程试剂盒包含一套完整的工具和试剂,用于从外周血中富集、扩增和重编程红细胞祖细胞。

亚型
专用培养基
 
细胞类型
造血干/祖细胞,多能干细胞
 
种属

 
应用
细胞培养,重编程
 
品牌
StemSpan,TeSR
 
研究领域
干细胞生物学
 
制剂类别
无血清
 

实验数据

Depletion of T-Cells and B-Cells From Whole Blood With RosetteSep™ Human Progenitor Cell Basic Pre-Enrichment Cocktail and SepMate™ Tubes

Figure 1. Depletion of T-Cells and B-Cells from Whole Blood with RosetteSep™ Human Progenitor Cell Basic Pre-Enrichment Cocktail and SepMate™ Tubes

(A) In the donor sample shown above, T-cells (CD3+) represent approximately 21.2% of the PBMC fraction while B-cells (CD19+) are present at 3.4%. (B) The addition of the RosetteSep™ Human Progenitor Cell Basic Pre-Enrichment cocktail efficiently depletes the T- and B-cell population to <1% of the enriched cell fraction.

Expansion of Erythroid Progenitor Cells Isolated From Peripheral Blood Using StemSpan™ SFEM II and StemSpan™ Erythroid Expansion Supplement

Figure 2. Expansion of Erythroid Progenitor Cells Isolated from Peripheral Blood Using StemSpan™ SFEM II and StemSpan™ Erythroid Expansion Supplement

TNC: total nucleated cell, Average shown in bold (range).

Erythroid Progenitor Cells are Expanded in StemSpan™ SFEM II With Erythroid Expansion Supplement

Figure 3. Erythroid Progenitor Cells Are Expanded in StemSpan™ SFEM II with Erythroid Expansion Supplement

(A) Isolated PBMCs were expanded for seven days and then examined by flow cytometry for erythroid progenitor cells, T-cells and B-cells. Representative plots illustrate that erythroid progenitor cells (GlyA+CD71+) are enriched after seven days, though some T-cells (CD3+) and B-cells (CD19+) remain. (B) Use of the RosetteSep™ cocktail to deplete lineage-committed cells leads to increased purity of the expanded erythroid progenitors and little/no contaminating lymphoid cells. Note: same donor sample used for A and B.

Schematic of ReproTeSR™ Reprogramming Timeline

Figure 4. Schematic of ReproTeSR™ Reprogramming Timeline

ReproTeSR™ is used during the entire induction phase of reprogramming (day 3 to 21). On days 3 and 5, ReproTeSR™ is added to StemSpan™ growth media (in a fed-batch manner) to facilitate attachment of transfected cells. Attached cells are further cultured in ReproTeSR™ with daily full media changes until putative iPS cell colonies emerge (days 21-28). iPS cell colonies can then be isolated and propagated in TeSR™ media. (mTeSR™1, TeSR™2, TeSR™-E8™).

Blood Cell Reprogramming Efficiencies Are Higher in ReproTeSR™ Medium Compared to in hESC Medium

Figure 5. Blood Cell Reprogramming Efficiencies Are Higher in ReproTeSR™ Medium Compared to in hESC Medium

Efficiency of reprogramming (A) erythroid cells, or (B) CD34+ cells using episomal reprogramming vectors is higher in ReproTeSR™ medium compared to in KOSR-containing hESC medium. Data shown are mean +/- SEM, erythroid cells n=4, CD34+ cells n=5.

Generation of iPS Cells From 1mL of Peripheral Blood

Figure 6. Generation of iPS Cells from 1mL of Peripheral Blood

Starting from 1mL of PB, PBMCs were enriched, erythroid progenitors were expanded and reprogrammed in ReproTeSR™. (A) Approximately 75 iPS-like colonies that were positive for alkaline phosphatase expression (blue) were generated. (B-C) iPS cell colonies exhibit compact ES-like morphology with defined borders and high nuclear to cytoplasmic ratio. Representative images of generated iPS cell colonies taken at 20X (B) and 400X (C) magnification are shown.

Reprogramming Efficiency of CD34+ and Erythroid Progenitor Cells With ReproTeSR™

Figure 7. Reprogramming Efficiency of CD34+ and Erythroid Progenitor Cells With ReproTeSR™

Average values in bold (range).

产品说明书及文档

请在《产品说明书》中查找相关支持信息和使用说明,或浏览下方更多实验方案。

Document Type
Product Name
Catalog #
Lot #
Language
Document Type
Technical Manual
Catalog #
05924
Lot #
All
Language
English
Document Type
Safety Data Sheet 1
Catalog #
05924
Lot #
All
Language
English
Document Type
Safety Data Sheet 2
Catalog #
05924
Lot #
All
Language
English
Document Type
Safety Data Sheet 3
Catalog #
05924
Lot #
All
Language
English
Document Type
Safety Data Sheet 4
Catalog #
05924
Lot #
All
Language
English
Document Type
Safety Data Sheet 5
Catalog #
05924
Lot #
All
Language
English

应用领域

本产品专为以下研究领域设计,适用于工作流程中的高亮阶段。探索这些工作流程,了解更多我们为各研究领域提供的其他配套产品。

相关材料与文献

技术资料 (4)

文献 (1)

Development of induced pluripotent stem cells from a patient with hypertrophic cardiomyopathy who carries the pathogenic myosin heavy chain 7 mutation p.Arg403Gln. M. Holliday et al. Stem cell research 2018

Abstract

Hypertrophic cardiomyopathy (HCM) is an inherited cardiomyopathy characterized by left ventricular hypertrophy ≥15 mm in the absence of loading conditions. HCM has a prevalence of up to one in 200, and can result in significant adverse outcomes including heart failure and sudden cardiac death. An induced pluripotent stem cell (iPSC) line was generated from peripheral blood mononuclear cells obtained from the whole blood of a 38-year-old female patient with HCM in which genetic testing identified the well-known pathogenic p.Arg403Gln mutation in myosin heavy chain 7. iPSCs express pluripotency markers, demonstrate trilineage differentiation capacity, and display a normal 46,XX female karyotype. This resource will allow further assessment of the pathophysiological development of HCM.

更多信息

更多信息
种属 Human
配方类别 Serum-Free
质量保证:

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