CD34+ Progenitor Reprogramming Kit

Integrated workflow containing tools to isolate and expand CD34+ progenitor cells from peripheral blood and their subsequent reprogramming to iPS cells

产品号 #05925_C

Integrated workflow containing tools to isolate and expand CD34+ progenitor cells from peripheral blood and their subsequent reprogramming to iPS cells

产品优势


  • Optimized for enrichment, expansion and reprogramming of CD34+ progenitor cells expanded from peripheral blood samples

  • Improved reprogramming efficiency and higher frequency of iPS cell colonies, compared to traditional hES cell medium

  • Rapid emergence of large colonies with high quality iPS cell-like morphology facilitates identification and subcloning

  • Seamlessly integrates with TeSR™ and STEMdiff™ products for downstream maintenance and differentiation of iPS cell lines

产品组分包括

  • SepMate™-50 (Catalog #15440)
  • Lymphoprep™ (Catalog #18060)
  • Complete Kit for Human Whole Blood CD34+ Cells (Catalog #15086)
    • RosetteSep™ Human Hematopoietic Progenitor Cell Enrichment Cocktail, 3 x 2 mL
    • EasySep™ Human CD34 Positive Selection Cocktail, 0.4 mL
    • EasySep™ Dextran RapidSpheres™ 50100, 1 mL
  • StemSpan™ SFEM II (Catalog #09605)
  • StemSpan™ CD34+ Expansion Supplement (Catalog #02691)
  • ReproTeSR™ (Catalog #05926)
    • TeSR™-E7™/ReproTeSR™ Basal Medium
    • ReproTeSR™ 25X Supplement

概述

The CD34+ Progenitor Reprogramming Kit contains an integrated set of tools and reagents to enrich, expand and reprogram CD34+ progenitor cells from human peripheral blood samples.

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
 

Data Figures

Typical Performance of the EasySep™ Complete Kit for Human Whole Blood CD34+ Cells

Figure 1. Typical Performance of the EasySep™ Complete Kit for Human Whole Blood CD34+ Cells

(A) In whole PB, the proportion of CD34+/CD45+ cells is typically very low (<1%). (B) In the RosetteSep™ enrichment step, unwanted cells are removed during density gradient centrifugation. This step enriches the population of CD34+/CD45+ cells (19.8%) in this example). (C) After positive selection of CD34+ cells using EasySep™, a highly pure population of CD34+/CD45+ cells (96.2% in this example) can be obtained. Values are reported as a percentage of viable CD45+ cells.

StemSpan™ SFEM II and CD34+ Expansion Supplement Support Expansion of CD34+ Cells

Figure 2. StemSpan™ SFEM II and CD34+ Expansion Supplement Support Expansion of CD34+ Cells

TNC: Total Nucleated Cell

Schematic of ReproTeSR™ Reprogramming Timeline

Figure 3. 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 4. 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.

ReproTeSR™ Generates iPS Cell Colonies With Superior Colony Morphology

Figure 5. ReproTeSR™ Generates iPS Cell Colonies With Superior Colony Morphology

Representative images of iPS cell colonies generated from isolated CD34+ progenitor cells using (A) ReproTeSR™ and (B) hESC) medium. iPS cell colonies produced using ReproTeSR™ exhibit more defined borders, compact morphology and reduced differentiation compared with hESC medium. 200X magnification.

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

Figure 6. 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
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Technical Manual
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05925
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All
Language
English
Document Type
Safety Data Sheet 1
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05925
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English
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Safety Data Sheet 2
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05925
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All
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English
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Safety Data Sheet 3
Catalog #
05925
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All
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English
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Safety Data Sheet 4
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05925
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All
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English
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Safety Data Sheet 5
Catalog #
05925
Lot #
All
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English
Document Type
Safety Data Sheet 6
Catalog #
05925
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.

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.
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