Use StemSpan™ Lymphoid Progenitor Expansion Supplement (10X) to selectively promote the expansion and differentiation of CD34+ cells isolated from human cord blood (CB) samples to CD7+CD5+ progenitor T (pro-T) cells.
Available either individually or as part of the StemSpan™ T Cell Generation Kit or STEMdiff™ T Cell Kit, this supplement is intended for use in combination with StemSpan™ SFEM II medium and plates coated with StemSpan™ Lymphoid Differentiation Coating Material (100X). For further maturation of pro-T cells to CD4+CD8+ double-positive (DP) T cells, you can use StemSpan™ T Cell Progenitor Maturation Supplement (10X).
For detailed information on protocols for expansion and differentiation of CD34+ cells with StemSpan™, please explore the Technical Manual and Educational Materials.
Available either individually or as part of the StemSpan™ T Cell Generation Kit or STEMdiff™ T Cell Kit, this supplement is intended for use in combination with StemSpan™ SFEM II medium and plates coated with StemSpan™ Lymphoid Differentiation Coating Material (100X). For further maturation of pro-T cells to CD4+CD8+ double-positive (DP) T cells, you can use StemSpan™ T Cell Progenitor Maturation Supplement (10X).
For detailed information on protocols for expansion and differentiation of CD34+ cells with StemSpan™, please explore the Technical Manual and Educational Materials.
Data Figures
Figure 1. Frequency and Yield of CD7+CD5+ Pro-T Cells After 14 Days of Culture
CB-derived CD34+ cells (freshly isolated or frozen) were cultured for 14 days in StemSpan™ SFEM II containing Lymphoid Progenitor Expansion Supplement (Catalog #09915) on plates coated with Lymphoid Differentiation Coating Material (Catalog #09925). Cells were harvested and analyzed for CD7 and CD5 expression by (A) flow cytometry. The (B) average frequency of viable CD7+CD5+ pro-T cells on day 14 was 70%, with ~200 CD7+CD5+ cells produced per input CD34+ cell. Shown are means with 95% confidence intervals (n = 33).
Figure 2. Frequency and Yield of CD4 ISP and CD4+CD8+ DP Cells After 42 Days of Culture
CB-derived CD34+ cells (freshly isolated or frozen) were cultured with the StemSpan™ T Cell Generation Kit (Catalog #09940) for 42 days and (A) analysed by flow cytometry for the expression of CD4, CD8, CD3 and TCRαβ. The (B) frequency and (C) yield of CD4 ISP, double-positive (CD4+CD8+) and CD3+TCRαβ+-expressing double-positive cells (CD4+CD8+CD3+TCRαβ+) are shown. On average, 38% of the total viable population were DP (CD4+CD8+), of which 35% co-expressed CD3 and TCRαβ. The yields of total DP cells and CD3+TCRαβ+ DP cells per input CD34+ cell were ~23,000 and ~9,000, respectively. Shown are means with 95% confidence intervals (n = 31).
Figure 3. Frequency and Yield of CD8 SP T Cells After 49 Days of Culture
DP cells were further matured into CD8 SP T cells by culturing for an additional 7 days in StemSpan™ SFEM II with T Cell Progenitor Maturation Supplement (Catalog #09930), IL-15 (Catalog #78031) and ImmunoCult™ CD3/CD28/CD2 T Cell Activator (Catalog #10970) on coated plates. On day 49, cells were (A) analyzed by flow cytometry for the expression of CD3, TCRαβ, CD4 and CD8. The (B) frequency and yield of CD3+TCRαβ+-expressing cells and their subsets are shown. On average, 54% of the CD3+TCRαβ+ cells were DP (CD4+CD8+) and 38% were CD8 SP (CD4-CD8+). The average yield of CD8 SP T cells per input CD34+ cell was ~6,000. CD3+TCRαβ+ CD4 SP (CD4+CD8-) T cells were detected at very low frequencies (data not shown). Shown are means with 95% confidence intervals (n = 12).
Protocols and Documentation
Find supporting information and directions for use in the Product Information Sheet or explore additional protocols below.
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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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