Erythroid Progenitor Reprogramming Kit

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

Erythroid Progenitor Reprogramming Kit

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

1 Kit
Catalog #05924
838 USD

Overview

The Erythroid Progenitor Reprogramming Kit contains an integrated set of tools and reagents to enrich, expand and reprogram erythroid progenitor cells from peripheral blood.
Advantages:
• Optimized for enrichment, expansion and reprogramming of erythroid 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
Components:
  • SepMate™-15 (Catalog #15410)
  • Lymphoprep™ (Catalog #07801)
  • RosetteSep™ Human Progenitor Cell Basic Pre-Enrichment Cocktail (Catalog #15216)
  • StemSpan™ SFEM II (Catalog #09605)
  • StemSpan™ Erythroid Expansion Supplement (Catalog #02692)
  • ReproTeSR™ (Catalog #05920)
    • ReproTeSR™ Basal Medium
    • ReproTeSR™ 20X Supplement
    • ReproTeSR™ 500X Supplement
Subtype:
Specialized Media
Cell Type:
Pluripotent Stem Cells; Hematopoietic Stem and Progenitor Cells
Species:
Human
Application:
Reprogramming; Cell Culture
Brand:
TeSR; StemSpan
Area of Interest:
Stem Cell Biology
Formulation:
Serum-Free; Defined

Scientific Resources

Educational Materials

(3)

Product 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.

Data and Publications

Data

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).

STEMCELL TECHNOLOGIES INC.’S QUALITY MANAGEMENT SYSTEM IS CERTIFIED TO ISO 13485. PRODUCTS ARE FOR RESEARCH USE ONLY AND NOT INTENDED FOR HUMAN OR ANIMAL DIAGNOSTIC OR THERAPEUTIC USES UNLESS OTHERWISE STATED.
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