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细胞培养基及添加物

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

概要
技术资料
数据及文献

概要

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.

数据及文献

Data

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).