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
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EasySep™ MagnetMagnet for column-free immunomagnetic separation
Overview
Data Figures
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.
Figure 2. StemSpan™ SFEM II and CD34+ Expansion Supplement Support Expansion of CD34+ Cells
TNC: Total Nucleated Cell
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™).
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.
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.
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.
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.
Resources and Publications
Educational Materials (4)
Related Products
Quality Statement:
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.
