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细胞培养基及添加物

StemSpan™ CD34+ Expansion Supplement (10X)

Serum-free culture supplement for expansion of human CD34+ hematopoietic cells

概要
技术资料
数据及文献

概要

StemSpan™ CD34+ Expansion Supplement (10X) contains a combination of recombinant human cytokines and other additives formulated to selectively promote the expansion of CD34+ cells isolated from human cord blood (CB) or bone marrow (BM) samples.

StemSpan™ CD34+ Expansion Supplement typically promotes ~40-fold expansion of total nucleated cells in 7-day liquid cultures of CD34+ human cord blood (CB) cells. After one week, ~40% of the cultured cells express CD34, indicative of >10-fold expansion of input CD34+ CB cells. This expansion may be further increased with the addition of small molecules such as UM729. See data tab for more details.

StemSpan™ CD34+ Expansion Supplement (10X) is intended for use in combination with any of the following StemSpan™ media:
• StemSpan™ SFEM (Catalog #09600)
• StemSpan™ SFEM II (Catalog #09605)
• StemSpan™-XF (Catalog #100-0073)
• StemSpan™-AOF (Catalog #100-0130)

技术资料

数据及文献

Data

Table 1. HSC Expansion Culture with CD34+ Human Cord Blood Cells Cultured in StemSpan™ SFEM Containing CD34+ Expansion Supplement

HSC Expansion Culture with CD34+ Human Cord Blood Cells Cultured in StemSpan™ SFEM Containing CD34+ Expansion Supplement

Shown are the percent CD34+ cells, fold expansion of total nucleated cells (TNC) and CD34+ cells, and numbers of colony-forming units (CFU) produced per input CD34+ cell after 7 days of hsc expansion culture of enriched CD34+ cells from six independent cord blood (CB) samples.
*95% confidence limits, the range within which 95% of the results will typically fall.
ND: not done

Comparison of HSC expansion in different StemSpan™ media containing CD34+ Expansion Supplement

Figure 1. Comparison of CD34+ Cell Expansion in Different StemSpan™ Media Containing CD34+ Expansion Supplement

Average expansion of (A) total nucleated cells (TNC), (B) CD34+ cells and (C) colony-forming units (CFU), normalized relative to the values obtained in StemSpan™ SFEM (grey bars) after culturing purified hematopoietic CD34+ cord blood cells (n=6) for 7 days in StemSpan™ SFEM, SFEM II (gold bars) or ACF (orange bars) media containing CD34+ Expansion Supplement. Vertical lines indicate 95% confidence limits, the range within which 95% of results will typically fall. Cell yields in StemSpan™ SFEM II were on average ~60% higher than in StemSpan™ SFEM and StemSpan™ ACF.
*p<0.001, #p<0.05 (paired t-test, n=6 in A and B, n=4 in C).

Note: Data for StemSpan™-ACF shown were generated with the original phenol red-containing version. However internal testing showed that the performance of the new phenol red-free, cGMP-manufactured version, StemSpan™-AOF (Catalog #100-0130) was comparable.

Publications (2)

Cell 2018 JAN Intrinsic Immunity Shapes Viral Resistance of Stem Cells. Wu X et al.

Abstract

Stem cells are highly resistant to viral infection compared to their differentiated progeny; however, the mechanism is mysterious. Here, we analyzed gene expression in mammalian stem cells and cells at various stages of differentiation. We find that, conserved across species, stem cells express a subset of genes previously classified as interferon (IFN) stimulated genes (ISGs) but that expression is intrinsic, as stem cells are refractory to interferon. This intrinsic ISG expression varies in a cell-type-specific manner, and many ISGs decrease upon differentiation, at which time cells become IFN responsive, allowing induction of a broad spectrum of ISGs by IFN signaling. Importantly, we show that intrinsically expressed ISGs protect stem cells against viral infection. We demonstrate the in vivo importance of intrinsic ISG expression for protecting stem cells and their differentiation potential during viral infection. These findings have intriguing implications for understanding stem cell biology and the evolution of pathogen resistance.
Nature methods 2017 JUN Marker-free coselection for CRISPR-driven genome editing in human cells. D. Agudelo et al.

Abstract

Targeted genome editing enables the creation of bona fide cellular models for biological research and may be applied to human cell-based therapies. Therefore, broadly applicable and versatile methods for increasing its efficacy in cell populations are highly desirable. We designed a simple and robust coselection strategy for enrichment of cells with either nuclease-driven nonhomologous end joining (NHEJ) or homology-directed repair (HDR) events by harnessing the multiplexing capabilities of CRISPR-Cas9 and Cpf1 systems. Selection for dominant alleles of the ubiquitous sodium/potassium pump (Na+/K+ ATPase) that rendered cells resistant to ouabain was used to enrich for custom genetic modifications at another unlinked locus of interest, thereby effectively increasing the recovery of engineered cells. The process is readily adaptable to transformed and primary cells, including hematopoietic stem and progenitor cells. The use of universal CRISPR reagents and a commercially available small-molecule inhibitor streamlines the incorporation of marker-free genetic changes in human cells.
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