StemSpan™-XF Xeno-Free Human Hematopoietic Cell Medium

总览

StemSpan™-XF专为体外培养和扩增人造血细胞而开发，可添加适当的生长因子和添加物。这使得用户可以根据实验需求灵活地配制培养基。StemSpan™-XF含有经过测试的人源性和重组人 (rh) 蛋白。

当搭配合适的StemSpan™扩增添加物，StemSpan™-XF可用于扩增从人脐带血、动员外周血或骨髓样本中分离的 CD34+细胞，或扩增和分化谱系定向祖细胞，从而生成红系、髓系或巨核细胞祖细胞群。

包含
本产品仅含有人衍生或重组人蛋白质。

分类
专用培养基

细胞类型
造血干/祖细胞

种属
人

应用
细胞培养，扩增

品牌
StemSpan

研究领域
干细胞生物学，移植研究

制剂类别
无血清，无异源

实验数据

Figure 1. Immunophenotyping of Cultured CD34⁺ Cells

Cord blood (CB)-derived CD34⁺ cells were purified using EasySep™ Human Cord Blood CD34 Positive Selection Kit II (Catalog #17896) and cultured in StemSpan™-XF (Catalog #100-0073) supplemented with StemSpan™ CD34⁺ Expansion Supplement (Catalog #02691) and UM171*. After 7 days, the cultured cells were stained with fluorescently labeled antibodies against CD34, CD90 and CD45RA, in addition to viability dye 7-AAD, and analyzed by flow cytometry. Sequential gates were used to determine the percentages of viable (A) CD34⁺ and CD34⁺CD90⁺CD45RA^- cells, based on fluorescence minus one (FMO) controls and (B) CD34^bright and CD34^brightCD90⁺CD45RA^- cells.

*Similar results are expected when using UM729 (Catalog #72332) prepared to a final concentration of 1μM. For more information including data comparing UM171 and UM729, see Fares et al. 2014.

Figure 2. StemSpan™ Media Support Greater Expansion of Human CD34⁺ and CD34^bright Cells Than Other Commercial Media

Purified CB-derived CD34⁺ cells were cultured for 7 days in StemSpan™ media (SFEM, SFEM II, AOF, and XF, orange bars), and in five media from other suppliers (Commercial Alternative, grey bars). All media were supplemented with StemSpan™ CD34⁺ Expansion Supplement and UM171*. The (A) frequency and (B) cell expansion of viable CD34⁺ and CD34^bright cells in culture were analyzed by flow cytometry as described in Figure 1. Compared to the commercial alternatives tested, StemSpan™ media showed significantly higher expansion of CD34⁺ and CD34^bright cells (P < 0.05 when comparing StemSpan™ SFEM, SFEM II, and XF to five media from other suppliers, calculated using a one-way ANOVA followed by Dunnett’s post hoc test). The CD34bright cell population is enriched for functional stem/progenitor cells (See Figure 4). Data shown are mean ± SEM (n = 8).

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.

*Similar results are expected when using UM729 (Catalog #72332) prepared to a final concentration of 1μM. For more information including data comparing UM171 and UM729, see Fares et al. 2014.

Figure 3. StemSpan™ Media Support Equal or Greater Expansion of Primitive Human CD34^brightCD90⁺CD45RA^- Cells Than Other Commercial Media

Purified CB-derived CD34⁺ cells were cultured for 7 days in StemSpan™ media (SFEM, SFEM II, AOF, and XF, orange bars), and in five media from other suppliers (Commercial Alternative, grey bars). All media were supplemented with StemSpan™ CD34⁺ Expansion Supplement and UM171*. The (A) frequency and (B) cell expansion of viable CD34⁺CD90⁺CD45RA^- (solid) and CD34^brightCD90⁺CD45RA^- (dotted overlay) cells in culture were analyzed by flow cytometry as described in Figure 1. Compared to the commercial alternatives tested, StemSpan™ media showed similar or significantly higher expansion of CD34^brightCD90⁺CD45RA^- cells (P < 0.05 when comparing StemSpan™ SFEM II and XF to five media from other suppliers, calculated using one-way ANOVA followed by Dunnett’s post hoc test). The CD34^brightCD90⁺CD45RA^- cell population is highly enriched for functional stem/progenitor cells (See Figure 4). Data shown are mean ± SEM (n = 8).

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.

*Similar results are expected when using UM729 (Catalog #72332) prepared to a final concentration of 1μM. For more information including data comparing UM171 and UM729, see Fares et al. 2014.

Figure 4. Culture-Expanded CD34^bright Cells Have Higher Colony-Forming Potential Than Culture-Expanded CD34^med and CD34^- Cells

Purified CB-derived CD34⁺ cells (n = 2) were cultured in StemSpan™-XF with StemSpan™ CD34⁺ Expansion Supplement and UM171* for 7 days. Cultured cells were stained with an anti-CD34 antibody and 7-AAD for sorting. Anti-CD90, and CD45RA antibodies were also included, to be used in later analyses. Cells were either (A) bulk-sorted or (B,C) single cell index-sorted from CD34^bright, CD34^med, and CD34^- populations. Sorted cells were plated in colony-forming unit (CFU) assays using MethoCult™ H4034 Optimum (Catalog #04034). (A) Bulk-sorted cells were plated at 400 cells/mL in 6-well SmartDish™ plates (Catalog #27370) and (B,C) single cells were sorted directly into 96-well plates (one cell per well, prefilled with 100 µL of MethoCult™). After 14 days of incubation, colonies were counted with (A) STEMvision™ or (B,C) manually using an inverted microscope. (A) Shown are representative images of CFU assays from bulk-sorted cells of CD34^bright, CD34^med, and CD34^- populations. The frequency of colonies in each population from the single cell-sorted experiments are shown as (B) CFU frequency, calculated as the total number of colonies generated by the total number of individually sorted cells (shown respectively as a fraction above each bar), and (C) CFU output expressed as total numbers of CFU in each sorted population per original input CD34⁺ cell.

The culture-expanded CD34^bright cells showed (B) higher CFU frequency (~50%) than CD34^med cells (~20%) and CD34^- cells (~10%), and yielded more diverse and primitive colony types, including CFU-GEMM and BFU-E colonies, which were rarely generated from CD34^med and CD34^- cells. When phenotyping data, collected during sorting, was analyzed all CFU-GEMM and the majority of of BFU-E colonies were generated from the fraction enriched for LT-HSCs, the CD34^brightCD90⁺CD45RA^- cell population, while 33% of BFU-E colonies were generated from the fraction enriched for ST-HSCs, the CD34^brightCD90-CD45RA- cell population. Only small CFU-G and CFU-M colonies were generated from CD34^brightCD90^+/-CD45RA⁺ cells, which is consistent with publications suggesting this population contains only progenitor cells.

*Similar results are expected when using UM729 (Catalog #72332) prepared to a final concentration of 1μM. For more information including data comparing UM171 and UM729, see Fares et al. 2014.

产品说明书及文档

请在《产品说明书》中查找相关支持信息和使用说明，或浏览下方更多实验方案。

Document Type

Product Name

Catalog #

Lot #

Language

Document Type

Product Information Sheet

Product Name

StemSpan™-XF

Catalog #

100-0073

Lot #

All

Language

English

Document Type

Safety Data Sheet

Product Name

StemSpan™-XF

Catalog #

100-0073

Lot #

All

Language

English

应用领域

本产品专为以下研究领域设计，适用于工作流程中的高亮阶段。探索这些工作流程，了解更多我们为各研究领域提供的其他配套产品。

Research Area

Workflow Stages

Workflow Stages for Hematopoietic Stem and Progenitor Cell Research

Cell Sourcing and Isolation

Expansion and Differentiation

Analysis

相关材料与文献

Modulation of β-Catenin promotes WNT expression in macrophages and mitigates intestinal injury R. M. Chugh et al. Cell Communication and Signaling : CCS 2025 Feb

Abstract

Macrophages are the major source of WNT ligands. However, the regulation of WNT expression in macrophages has not been studied. In the present study, we have discovered that activation of canonical β-Catenin signaling suppresses WNT expression in macrophages. EVs from these pre-conditioned macrophages promoted intestinal stem cell regeneration and mitigated intestinal injury. ChIP-seq analysis and validation studies using recombinant DNA construct expressing Luciferase reporter under WNT promoter (e.g. WNT5a and WNT9b) were conducted to demonstrate the involvement of β-Catenin in the transcriptional regulation of WNT expression. The regulatory role of β-Catenin in WNT expression in macrophages was examined by treating these cells with a Tankyrase inhibitor. In addition, the gene expressing β-Catenin was deleted in macrophages using Csf1r.iCre; Ctnnb1 fl/fl mice model. Both pharmacological and genetically modulated macrophages were examined for WNT expression and activity by qPCR and TCF/LEF luciferase assay respectively. Additionally, Csf1r.iCre; Ctnnb1 fl/fl mice were exposed to irradiation to compare the radiosensitivity with their wildtype littermate. Extracellular vesicles (EVs) were isolated from pre-conditioned WNT-enriched macrophages and infused in irradiated C57BL/6 and Lgr5/eGFP-IRES-Cre-ERT2 ; R26-ACTB-tdTomato-EGFP mice to determine the regenerative response of intestinal stem cell (ISC) and epithelial repair. Regenerative effects of EVs were also examined in mice model DSS induced colitis. ChIP-seq analysis and subsequent validation study suggested physical association of β-Catenin with WNT promoters to suppress WNT expression. Macrophage specific deletion of gene expressing β-Catenin or pharmacological inhibition of Tankyrase improves the WNT expression in macrophages several folds compared to control. Transfusion of these preconditioned macrophages or EVs from these cells delivers optimum level of morphogenic WNT to injured epithelium, activates ISC regeneration and mitigated radiation induced intestinal injury. Intestinal epithelium in Csf1r.iCre; Ctnnb1 fl/fl mice also showed radioresistance compared to wild type littermate. Moreover, EVs derived from WNT enriched macrophages can mitigate intestinal injury in mice model of DSS induced acute colitis. The study provides substantial evidence that macrophage-targeted modulation of canonical WNT signaling induces WNT expression in macrophages. Treatment with preconditioned macrophage derived WNT-enriched EVs can be a promising therapeutic approach against intestinal injury. The online version contains supplementary material available at 10.1186/s12964-025-02065-7.

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物种	人类
Contains	This product contains only human-derived or recombinant human proteins.
配方	无血清

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