StemSpan™-XF

Xeno-free and serum-free medium for culture and expansion of human hematopoietic cells
StemSpan™-XF

Xeno-free and serum-free medium for culture and expansion of human hematopoietic cells

500 mL Bottle
Catalog # 100-0073
490 USD

Overview

StemSpan™-XF has been developed for the in vitro culture and expansion of human hematopoietic cells, when the appropriate growth factors and supplements are added. This allows users the flexibility to prepare medium that meets their requirements. StemSpan™-XF contains pre-tested human-derived and recombinant human (rh) proteins.

Using appropriate StemSpan™ Expansion Supplements, StemSpan™-XF may be used to expand CD34+ cells isolated from human cord blood, mobilized peripheral blood, or bone marrow samples, or to expand and differentiate lineage-committed progenitor cells to generate populations of erythroid, myeloid, or megakaryocyte progenitor cells.
Components
  • StemSpan™-XF (Catalog #100-0073)
    • StemSpan™-XF, 500 mL
Contains
This product contains only human-derived or recombinant human proteins.
Subtype
Specialized Media
Cell Type
Hematopoietic Stem and Progenitor Cells
Species
Human
Application
Cell Culture, Expansion
Brand
StemSpan
Area of Interest
Stem Cell Biology, Transplantation Research
Formulation
Serum-Free, Xeno-Free

Scientific Resources

Product Documentation

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

Educational Materials (4)

Brochure
StemSpan™: Defined Media and Supplements for Hematopoietic Cell Expansion
Brochure
Hematopoietic Stem and Progenitor Cells - Products for Your Research
Webinar
New Tools for the Ex Vivo Expansion of Human Hematopoietic Stem and Progenitor Cells
1:06:52
New Tools for the Ex Vivo Expansion of Human Hematopoietic Stem and Progenitor Cells
Mini Review
Hematopoietic Stem and Progenitor Cells (HSPCs): Isolation, Culture, and Assays

Product 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.

Data and Publications

Data

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) CD34bright and CD34brightCD90+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 CD34bright 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 CD34bright 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 CD34bright 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 CD34brightCD90+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 CD34brightCD90+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 CD34brightCD90+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 CD34brightCD90+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 CD34bright Cells Have Higher Colony-Forming Potential Than Culture-Expanded CD34med 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 CD34bright, CD34med, 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 CD34bright, CD34med, 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 CD34bright cells showed (B) higher CFU frequency (~50%) than CD34med 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 CD34med 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 CD34brightCD90+CD45RA- cell population, while 33% of BFU-E colonies were generated from the fraction enriched for ST-HSCs, the CD34brightCD90-CD45RA- cell population. Only small CFU-G and CFU-M colonies were generated from CD34brightCD90+/-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.

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