StemSpan™ T Cell Generation Kit

For expansion and differentiation of human CD34+ hematopoietic progenitor cells to T cells

StemSpan™ T Cell Generation Kit

For expansion and differentiation of human CD34+ hematopoietic progenitor cells to T cells

StemSpan™ T Cell Generation Kit
1 Kit
1,431 USD
Catalog # 09940

For expansion and differentiation of human CD34+ hematopoietic progenitor cells to T cells

INDIVIDUAL COMPONENTS

StemSpan™ SFEM II

162 USD
Catalog # 09605

Serum-free medium for culture and expansion of hematopoietic cells

100 mL

StemSpan™ Lymphoid Progenitor Expansion Supplement (10X)

317 USD
Catalog # 09915

Supplement for the expansion and differentiation of lymphoid progenitor cells

5 mL

StemSpan™ Lymphoid Differentiation Coating Material (100X)

246 USD
Catalog # 09925

Coating material for the differentiation of lymphoid cells

0.25 mL

StemSpan™ T Cell Progenitor Maturation Supplement (10X)

317 USD
Catalog # 09930

Supplement for the maturation of T cell progenitors to T cells

12.5 mL

What's Included

• StemSpan™ SFEM II, 2 x 100 mL
• StemSpan™ Lymphoid Progenitor Expansion Supplement (10X), 5 mL
• StemSpan™ Lymphoid Differentiation Coating Material (100X), 2 x 0.25 mL
• StemSpan™ T Cell Progenitor Maturation Supplement (10X), 12.5 mL

Overview

Use this two-step protocol to reproducibly expand thousands of CD4+CD8+ double-positive (DP) T cells in cultures initiated with CD34+ human cord blood (CB) cells.

In step one, CD34+ CB cells are cultured for 14 days in medium containing expansion supplement to promote their proliferation and differentiation into CD7+CD5+ pro-T cells. In step two, pro-T cells generated during those first 14 days are cultured for another 4 weeks in medium containing maturation supplement to promote their maturation into DP cells. This product is serum- and feeder-free, which helps eliminate variation introduced by serum and stromal cell lines.

StemSpan™ Lymphoid Progenitor Expansion Supplement (10X) contains a combination of recombinant human cytokines and other additives formulated to selectively promote the expansion and differentiation of CD34+ cells isolated from human CB samples to CD7+CD5+ progenitor T (pro-T) cells. Additionally, StemSpan™ T Cell Progenitor Maturation Supplement (10X) enables further maturation of pro-T cells to DP cells. It also supports maturation of DP cells to CD8 SP T cells with additional stimuli. StemSpan™ Lymphoid Progenitor Expansion Supplement (10X) and StemSpan™ T Cell Progenitor Maturation Supplement (10X) are intended for use in combination with StemSpan™ SFEM II medium (Catalog # 09605), and on plates coated with StemSpan™ Lymphoid Differentiation Coating Material (100X).

An optional protocol is available to further mature DP cells to CD8 SP T cells. This extended protocol uses complete T cell progenitor maturation medium made with reagents included in the StemSpan™ T Cell Generation Kit and which must be combined with ImmunoCult™ CD3/CD28/CD2 T Cell Activator (Catalog #10970 or #10971) and IL-15 (Catalog #78031).
Subtype
Specialized Media, Supplements
Cell Type
Hematopoietic Stem and Progenitor Cells, T Cells
Species
Human
Application
Cell Culture, Differentiation, Expansion
Brand
StemSpan
Area of Interest
Cancer, Cell Therapy Research, Immunology, Stem Cell Biology
Formulation
Serum-Free

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.

Document Type
Product Name
Catalog #
Lot #
Language
Catalog #
09940
Lot #
All
Language
English
Catalog #
09605
Lot #
All
Language
English
Catalog #
09915
Lot #
All
Language
English
Catalog #
09925
Lot #
All
Language
English
Catalog #
09930
Lot #
All
Language
English
Document Type
Technical Manual
Catalog #
09940
Lot #
All
Language
English
Document Type
Safety Data Sheet 1
Catalog #
09940
Lot #
All
Language
English
Document Type
Safety Data Sheet 2
Catalog #
09940
Lot #
All
Language
English
Document Type
Safety Data Sheet 3
Catalog #
09940
Lot #
All
Language
English
Document Type
Safety Data Sheet 4
Catalog #
09940
Lot #
All
Language
English
Document Type
Safety Data Sheet
Catalog #
09605
Lot #
All
Language
English
Document Type
Safety Data Sheet
Catalog #
09925
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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