StemSpan™ CD34+ Expansion Supplement (10X)

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

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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)

Data Figures

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 (blue bars) or AOF (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™-AOF. *p<0.001, #p<0.05 (paired t-test, n=6 in A and B, n=4 in C).

Note: Data for StemSpan™-AOF shown were generated with the original phenol red-containing version StemSpan™-ACF (Catalog #09855). However internal testing showed that the performance of the new phenol red-free, cGMP-manufactured version, StemSpan™-AOF (Catalog #100-0130) was comparable.

Protocols and Documentation

Find supporting information and directions for use in the Product Information Sheet or explore additional protocols below.

Document Type
Product Name
Catalog #
Lot #
Language
Catalog #
02691
Lot #
All
Language
English
Document Type
Safety Data Sheet
Catalog #
02691
Lot #
All
Language
English

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 (9)

Publications (2)

Isolation and Culture of Non-adherent Cells for Cell Reprogramming. Andrianto et al. Journal of stem cells & regenerative medicine 2022

Abstract

Coronary heart disease (CHD) is a leading cause of death globally, while its current management is limited to reducing the myocardial infarction area without actually replacing dead cardiomyocytes. Direct cell reprogramming is a method of cellular cardiomyoplasty which aims for myocardial tissue regeneration, and CD34+ cells are one of the potential sources due to their shared embryonic origin with cardiomyocytes. However, the isolation and culture of non-adherent CD34+ cells is crucial to obtain adequate cells for high-efficiency genetic modification. This study aimed to investigate the optimal method for isolation and culture of CD34+ peripheral blood cells using certain culture media. A peripheral blood sample was obtained from a healthy subject and underwent pre-enrichment, isolation, and expansion. The culture was subsequently observed for their viability, adherence, and confluence. Day 0 observation of the culture showed a healthy CD34+ cell with a round cell shape, without any adherent cells present yet. Day 4 of observation showed that CD34+ cells within the blood plasma medium became adherent, indicated by their transformations into spindle or oval morphologies. Meanwhile, CD34+ cells in vitronectin and fibronectin media showed no adherent cells and many of them died. Day 7 observation revealed more adherent CD34+ cells in blood plasma medium, and which had 75% of confluence. In conclusion, the CD34+ cells that were isolated using a combination of density and magnetic methods may be viable and adequately adhere in culture using blood plasma medium, but not in cultures using fibronectin and vitronectin.
Intrinsic Immunity Shapes Viral Resistance of Stem Cells. Wu X et al. Cell 2018 JAN

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.
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