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 Expansion Supplements

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

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

Expansion of CD34<sup>+</sup> Cells

Application:

Culture and expansion of human CD34+ hematopoietic cells

Differentiation of Erythroid, Myeloid and Megakaryocyte Lineage Cells

Differentiation into Erythroid 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

Differentiation into Megakaryocytes

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

Generation of Mature Blood Cells in Vitro

SFEM
Leberbauer et al. (2005) Different steroids co-regulate long-term expansion versus terminal differentiation in primary human erythroid progenitors. Blood 105(1)

SFEM II
Huijskens et al. (2014) Technical advance: ascorbic acid induces development of double-positive T cells from human hematopoietic stem cells in the absence of stromal cells. J Leukoc Biol 96(6)

CC100
Kumkhaek et al. (2013) MASL1 induces erythroid differentiation in human erythropoietin-dependent CD34+ cells through the Raf/MEK/ERK pathway. Blood 121(16)

CC110
Gaikwad et al. (2007) In Vitro Expansion of Erythroid Progenitors from Polycythemia Vera Patients Leads to Decrease in JAK2V617F Allele. Exp Hema 35(4)

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

SFEM
Delaney et al. (2010) Notch-mediated expansion of human cord blood progenitor cells capable of rapid myeloid reconstitution. Nat Med 16(2)
Cutler et al. (2013) Prostaglandin-modulated umbilical cord blood hematopoietic stem cell transplantation. Blood 122(17)

Generating Target Cells for Reprogramming to Make Induced Pluripotent Stem Cells

H3000 + CC100
Ohmine et al. (2011) Induced pluripotent stem cells from GMP-grade hematopoietic progenitor cells and mononuclear myeloid cells. Stem Cell Res and Therapy 2(6)

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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