CD34+祖细胞重编程试剂盒

包含从外周血中分离和扩增CD34+祖细胞及将其重编程为iPS细胞的工作流程

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产品号 #（选择产品）

产品号 #05925_C

包含从外周血中分离和扩增CD34+祖细胞及将其重编程为iPS细胞的工作流程

产品优势

优化了从外周血扩增的CD34+祖细胞的富集、扩增和重编程；
与传统的人胚胎干细胞（hES）培养基相比，iPS细胞重编程效率更高，iPS细胞克隆形成频率更高；
可快速形成具有高质量iPS细胞样形态的，有利于鉴定和亚克隆；
与TeSR™和STEMdiff™产品无缝衔接，用于iPS细胞系的下游维持和分化。

产品组分包括

SepMate™-50（产品号：#15440）；
Lymphoprep™（产品号：#18060）；
人全血CD34+细胞分选试剂盒（产品号：#15086）；
RosetteSep™人造血祖细胞富集抗体混合物，3 × 2ml；
EasySep™Human CD34 正选抗体混合物， 0.4 mL；
EasySep™Dextran RapidSpheres™50100磁珠，1 mL；
StemSpan™SFEM II（产品号：#09605）；
StemSpan™CD34+扩增添加物（产品号：#02691）；
ReproTeSR™（产品号：#05926）；
TeSR™-E7™/ReproTeSR™基础培养基；
ReproTeSR™25X添加物

实验数据

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.