StemSpan™-AOF

cGMP medium, for culture and expansion of human hematopoietic cells

StemSpan™-AOF

cGMP medium, for culture and expansion of human hematopoietic cells

StemSpan™-AOF
500 mL Bottle
552 GBP
Catalog # 100-0130

For culture and expansion of human hematopoietic cells

Product Advantages


  • 20X expansion of human cord blood-derived CD34+ cells after 7 days of culture

  • Maintenance of primitive CD34
  • Suitable for use in genome editing protocols

  • Animal origin- and phenol red-free formulation

  • Manufactured and tested under relevant cGMPs

  • Full traceability of raw materials

Overview

StemSpan™-AOF is an animal origin-free (AOF) medium that has been developed for the in vitro culture and expansion of human hematopoietic cells, when appropriate growth factors/cytokines and supplements (e.g. small molecules) are added. This allows users the flexibility to prepare medium that meets their requirements.

StemSpan™-AOF contains only recombinant proteins and synthetic components, and does not contain serum or other human- or animal-derived components.

Using appropriate cytokines, StemSpan™-AOF 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 myeloid or megakaryocyte progenitor cells.

Please note, StemSpan™-AOF was originally launched as StemSpan™-ACF Without Phenol Red. This name change signifies that in addition to being animal component-free, no materials of animal or human origin are used in the manufacture of this medium or its components, to at least the secondary level of manufacturing. This medium also replaces StemSpan™-ACF (Catalog #09855).

StemSpan™-AOF (Catalog #100-0130) is manufactured under relevant cGMPs, ensuring the highest quality and consistency for reproducible results. For additional quality information, visit www.STEMCELL.com/compliance.

Request a Letter of Authorization (LOA) for StemSpan™-AOF's Drug Master File.
Contains
This product contains only recombinant proteins and synthetic components.
Subtype
Specialized Media
Cell Type
Hematopoietic Stem and Progenitor Cells
Species
Human
Application
Cell Culture, Differentiation, Expansion, Genome Editing
Brand
StemSpan
Area of Interest
Cell Therapy Research, Stem Cell Biology, Transplantation Research
Formulation
Animal Origin-Free, Serum-Free, Xeno-Free

Data Figures

Day 7 Immunophenotyping of CD34+ Cells Cultured in StemSpan™-AOF

Figure 1. Day 7 Immunophenotyping of CD34+ Cells Cultured in StemSpan™-AOF

CD34+ cells were purified from cord blood (CB) using the EasySep™ Human Cord Blood CD34 Positive Selection Kit II (Catalog #17896) and cultured in StemSpan™-AOF (Catalog #100-0130) supplemented with StemSpan™ CD34+ Expansion Supplement (Catalog #02691) (A) without or (B) with the addition of UM729 (Catalog #72332). 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. The horizontal dotted line in the CD34 vs FSC plots indicates the boundary between CD34- and CD34+ cells as based on a fluorochrome minus one (FMO) control for CD34 expression. Orange gates on these plots indicate the population of CD34bright cells used to generate data in Figures 2 and 3. Sequential gates were used to determine the percentages of viable CD34+ cells, CD34bright cells, and CD34brightCD90+CD45RA- cells.

StemSpan™-AOF Supports Equivalent or Greater Expansion of Human CD34+ and CD34bright Cells than Other Commercial Media

Figure 2. StemSpan™-AOF Supports Equivalent or Greater Expansion of Human CD34+ and CD34bright Cells than Other Commercial Media

Purified cord blood (CB)-derived CD34+ cells were cultured for 7 days in StemSpan™-AOF (orange bar), and in four xeno-free media formulations from other suppliers (Xeno-Free Commercial Alternative, grey bars), including (in random order) SCGM (Cellgenix), X-VIVO™ 15 (Lonza), Stemline™ II (Sigma), and StemPro™-34 (Thermo). The (A) frequency and (B) cell expansion of viable CD34+ and CD34bright cells in culture were based on viable cell counts and flow cytometry results. StemSpan™-AOF, the only animal origin-free formulation, showed equivalent performance to all xeno-free alternative media tested. All media were supplemented with StemSpan™ CD34⁺ Expansion Supplement and UM171*. 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.

StemSpan™ Media Support Greater Expansion of Human CD34+CD90+CD45RA- and CD34brightCD90+CD45RA- Cells than Other Commercial Media

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™-AOF medium (orange bar), and in four xeno-free media formulations from other suppliers (Commercial Alternative, gray bars) including (in random order) SCGM (Cellgenix), X-VIVO 15 (Lonza), Stemline II (Sigma), and StemPro 34 (Thermo). All media were supplemented with StemSpan™ CD34+ Expansion Supplement and UM171*. The (A) frequency and (B) cell expansion of CD34+CD90+CD45RA- (solid) and CD34brightCD90+CD45RA-(dotted overlay) cells in culture were based on viable cell counts and flow cytometry results as shown in Figure 1. StemSpan™-AOF showed similar or significantly higher expansion of CD34brightCD90+CD45RA- cells (P < 0.05 compared to four media from other suppliers, calculated using one-way ANOVA followed by Dunnett’s post hoc test) and equivalent performance to all xeno-free competitors tested. 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.

StemSpan™ Media Support Better CD34+ and Primitive CD34+CD90+CD45RA- HSPC Expansion for Genome Editing Applications Compared with Alternative Commercial Media

Figure 4. StemSpan™ Media Support Better CD34+ and Primitive CD34+CD90+CD45RA- HSPC Expansion in a Genome Editing Application Compared with Alternative Commercial Media

Purified CB-derived CD34+ cells were cultured for 2 days in select StemSpan™-AOF, (orange bar), or four xeno-free media formulations from other suppliers (gray bars). All media were supplemented with StemSpan™ CD34+ Expansion Supplement and UM171*. Cells were then electroporated using Arcitect™ CRISPR-Cas9 RNP complexes containing crRNA:tracrRNA targeting beta-2-microglobulin (B2M), and cultured for an additional 4 days in the same conditions. Knockout efficiency as measured by staining for MHC-I and analyzing by flow cytometry, was similar in all media tested, ~70-80%. (A) The percentage of CD34+ cells and (B) CD34+CD90+CD45RA- cells were quantified by flow cytometry 4 days post-electroporation. Data shown are mean + SD (n = 4 donors; **P < 0.01).

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

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

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
Product Name
StemSpan™-AOF
Catalog #
100-0130
Lot #
All
Language
English
Document Type
Safety Data Sheet
Product Name
StemSpan™-AOF
Catalog #
100-0130
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.

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