StemSpan™

Hematopoietic Cell Expansion Media and Supplements

StemSpan™ serum-free hematopoietic cell expansion media promote the expansion and/or lineage-specific differentiation of human hematopoietic stem and progenitor cells (HSPCs) when supplemented with hematopoietic growth factors and/or other stimuli selected by the user. StemSpan™ Expansion Supplements are pre-mixed cocktails of recombinant human cytokines and other additives formulated to selectively promote the expansion of CD34+ stem and progenitor cells, or to stimulate their differentiation into erythroid, myeloid or megakaryocyte progenitors, when added to StemSpan™ media. A complete kit for the expansion and lineage-specific differentiation of CD34+ cells into T cell progenitors in stroma-free conditions is also available for research into lymphopoiesis.

Why Use StemSpan™ Media and Expansion Supplements?

  • Components are carefully selected to minimize lot-to-lot variability and support optimal and consistent culture results.
  • Media do not contain cytokines, allowing the flexibility to add StemSpan™ Expansion Supplements, cytokines and/or additives.
  • StemSpan™ SFEM II, combined with the appropriate Expansion Supplement, supports greater expansion of CD34+ cells and differentiation of erythroid cells, myeloid cells, megakaryocytes and T cell progenitors than other media tested.
  • StemSpan™-ACF is the first commercially available animal component-free medium for culturing HSPCs.

Expansion Media

StemSpan™ SFEM

StemSpan™ SFEM

Formulation:

Serum-free

Species:

  • Human
  • Mouse
  • Rat
  • Non-human primate

Recommended for:

Culture of HSPCs in serum-free conditions

Components:

  • BSA
  • Insulin
  • Tranferrin
  • Supplements
  • IMDM base

StemSpan™ SFEM II

StemSpan™ SFEM II

Formulation:

Serum-free

Species:

Human

Recommended for:

Culture of HSPCs in serum-free conditions

Components:

  • BSA
  • Insulin
  • Tranferrin
  • Supplements
  • IMDM base

StemSpan™ H3000

StemSpan™ H3000

Formulation:

Xeno-Free

Species:

Human

Recommended for:

Culture of HSPCs in the absence of non-human animal-derived components

Components:

  • Human-derived and recombinant human proteins
  • IMDM base

ACF StemSpan™ Media

Formulation:

Animal-Component Free

Species:

Human

Recommended for:

Culture of HSPCs in the absence of human- or animal-derived components

StemSpan™-ACF
  • Expansion of CD34+ cells and differentiation of myeloid cells and megakaryocytes

StemSpan™ Erythroid Expansion Medium (ACF-E)
  • Differentiation of erythroid progenitor cells

Components:

  • Recombinant and synthetic components
  • IMDM base
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Expansion Supplements for Human HSPCs

Expansion of CD34+ Cells

Application:

Culture and expansion of human CD34+ hematopoietic cells

Differentiation of Erythroid, Myeloid and Megakaryocyte Lineage Cells

Application:

Expansion and lineage-specific differentiation of human CD34+ cells to generate large numbers of erythroid progenitors, myeloid progenitors or megakaryocytes

Differentiation of Lymphoid Lineage Progenitor Cells

Application:

Expansion and lineage-specific differentiation of human CD34+ cells to generate large numbers of T cell progenitors in stroma-free conditions
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Data

Figure 1. StemSpan™ SFEM II Serum-Free Expansion Medium Containing CC100 Cytokine Cocktail Supports Greater Expansion of Human CD34+ Cells Than Other Media Tested

Expansion of CD34+ cells normalized relative to the values obtained in StemSpan™ SFEM medium (dark gray bar) after culturing purified CD34+ CB cells for 7 days in StemSpan™ SFEM, SFEM II (gold bar) and ACF (orange bar), and six media from other commercial suppliers (light gray bars; Competitor 1-6, which included, in random order, StemLine II (Sigma), HPGM (Lonza), HP01 (Macopharma), SCGM (Cellgenix), StemPro34 (Life Technologies) and X-Vivo-15 (Lonza)). All media were supplemented with StemSpan™ CC100 Cytokine Cocktail. Vertical lines indicate 95% confidence limits, the range within which 95% of results fall. The numbers of cells produced in StemSpan™ SFEM II were significantly higher than in all other media, except the numbers of CB cells produced in StemSpan™-ACF (*p<0.05, paired t-test, n=6).



Figure 2. StemSpan™ SFEM II Serum-Free Expansion Medium Containing CD34+ Expansion Supplement Supports Greater Expansion of Human CD34+ Cells Than Other Media Tested

Expansion of CD34+ cells normalized relative to the values obtained in SFEM medium (dark gray bar) after culturing purified CD34+ CB cells for 7 days in StemSpan™ SFEM, SFEM II (gold bar) and ACF (orange bar), and six media from other suppliers (light gray bars; Competitor 1-6, which included, in random order, StemLine II (Sigma), HPGM (Lonza), HP01 (Macopharma), SCGM (Cellgenix), StemPro34 (Life Technologies) and X-Vivo-15 (Lonza)). All media were supplemented with the StemSpan™ CD34+ Expansion Supplement. Vertical lines indicate 95% confidence limits, the range within which 95% of results fall. The numbers of cells produced in StemSpan™ SFEM II were significantly higher than in all other media (*p<0.01, paired t-test, n=6).



Figure 3. StemSpan™ SFEM II Serum-Free Expansion Medium Containing Erythroid Expansion Supplement Supports Greater Expansion of Erythroid Cells Than Other Media Tested

The numbers of erythroid cells, normalized relative to the values obtained in StemSpan™ SFEM medium (dark gray bar), obtained after culturing purified CD34+ CB cells for 14 days in StemSpan™ SFEM, SFEM II (gold bar) and ACF (orange bar), and six media from other commercial suppliers (light gray bars; Competitor 1-6, which included, in random order, StemLine II (Sigma), HPGM (Lonza), HP01 (Macopharma), SCGM (Cellgenix), StemPro34 (Life Technologies) and X-Vivo-15 (Lonza)). All media were supplemented with StemSpan™ Erythroid Expansion Supplement. Vertical lines indicate 95% confidence limits, the range within which 95% of results fall. The numbers of cells produced in StemSpan™ SFEM II were significantly higher than in all other media (*p<0.05, paired t-test, n=6).



Figure 4. StemSpan™ SFEM II Serum-Free Expansion Medium Containing Megakaryocyte Expansion Supplement Supports Greater Expansion of Megakaryocytes Than Other Media Tested

The numbers of megakaryocytes, normalized relative to the values obtained in StemSpan™ SFEM medium (dark gray bar), obtained after culturing purified CD34+ CB cells for 14 days in StemSpan™ SFEM, SFEM II (gold bar) and ACF (orange bar), and six media from other commercial suppliers (light gray bars; Competitor 1-6, which included, in random order, StemLine II (Sigma), HPGM (Lonza), HP01 (Macopharma), SCGM (Cellgenix), StemPro34 (Life Technologies) and X-Vivo-15 (Lonza)). All media were supplemented with StemSpan™ Megakaryocyte Expansion Supplement (Catalog #02696). Vertical lines indicate 95% confidence limits, the range within which 95% of results fall. The numbers of cells produced in the StemSpan™ media were significantly higher than in the other media (*p<0.01 paired t-test, n=6).



Table 1. Production of Myeloid Cells from Human CB-Derived CD34+ Cells Cultured in StemSpan™ SFEM II Containing Myeloid Expansion Supplement

Number of total nucleated cells (TNCs) produced per input human CB-derived CD34+ cell from 14 different donors and percentage of cells positive for myeloid markers CD13, CD14 and CD15 produced after 14-day culture in SFEM II containing Myeloid Expansion Supplement. Best expansion was obtained in SFEM II, but myeloid cell expansion may also be obtained in SFEM and ACF media. (*95% confidence limits (CL); the range within which 95% of the results will typically fall).



Figure 5. The StemSpan™ T Cell Progenitor Differentiation Kit Promotes the Expansion and Differentiation of CB-Derived CD34+ Cells into Pro- and Pre-T Cells

The average (A,C) frequencies and (B,D) numbers of (A,B) CD7+CD5+ pro-T cells and (C,D) CD7+CD1a+ pre-T cells on days 7, 14 and 21 of culture are shown for 10 - 26 independent experiments. The average frequency of (A) pro- and (C) pre-T cells were 84% and 28% respectively, after 21 days of culture. The number of (B) CD7+CD5+ pro-T cells increased (on average) ~10 - 100-fold every week, resulting in an average number of ~2100 pro-T cells produced per input CD34+ cell on day 21. After 21 days of culture (D) pre-T cells expressing CD7 and CD1a are present in large numbers, indicating the further differentiation of pro-T cells. The yield of (D) CD7+CD1a+ cells on day 21 was ~800 per input CD34+ cell. Vertical lines indicate 95% confidence limits (CL), the range within which 95% of results typically fall.



Key Applications

Ex Vivo Expansion of HSPCs for Rapid/Sustained Hematopoietic Recovery Post Transplantation

Generating Target Cells for Reprogramming to Make Induced Pluripotent Stem Cells

Gene Transfer into HSPCs

SFEM
Lechman et al. (2012) Attenuation of miR-126 Activity Expands HSC In Vivo without Exhaustion. Cell Stem Cell 11(6)

SFEM II
Buechele C et al. (2015) MLL leukemia induction by genome editing of human CD34+ hematopoietic cells. Blood. Epub ahead of print
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