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红细胞祖细胞重编程试剂盒

包含从外周血中分离和扩增红细胞祖细胞及其随后重编程为iPS细胞的集成工作流程
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产品号 #05924_C

包含从外周血中分离和扩增红细胞祖细胞及其随后重编程为iPS细胞的集成工作流程

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
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概述

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。

Subtype
Specialized Media
 
Cell Type
Hematopoietic Stem and Progenitor Cells, Pluripotent Stem Cells
 
Species
Human
 
Application
Cell Culture, Reprogramming
 
Brand
StemSpan, TeSR
 
Area of Interest
Stem Cell Biology
 
Formulation Category
Serum-Free
 

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Data Figures

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).

Protocols and Documentation

Find supporting information and directions for use in the Product Information Sheet or explore additional protocols below.

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

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.

Research Area
Workflow Stages

Resources and Publications

Educational Materials (4)

An Integrated Workflow for Reprogramming Blood Cells
Brochure
An Integrated Workflow for Reprogramming Blood Cells
hPSC Differentiation: Tools for Pluripotent Stem Cell-Derived Research
Brochure
hPSC Differentiation: Tools for Pluripotent Stem Cell-Derived Research
Products for Human Pluripotent Stem Cells
Brochure
Products for Human Pluripotent Stem Cells
Defined and Xeno-Free Medium for Reprogramming Blood-Derived CD34+ Cells or Erythroid Cells
Scientific Poster
Defined and Xeno-Free Medium for Reprogramming Blood-Derived CD34+ Cells or Erythroid Cells

Publications (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.
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Species Human
Formulation Category Serum-Free
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