StemSpan™ Lymphoid Differentiation Coating Material (100X)

Coating material for the expansion and differentiation of lymphoid cells

StemSpan™ Lymphoid Differentiation Coating Material (100X)

Coating material for the expansion and differentiation of lymphoid cells

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Coating material for the expansion and differentiation of lymphoid cells
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Overview

Use StemSpan™ Lymphoid Differentiation Coating Material (100X) to coat plates for the expansion and differentiation of human CD34+ hematopoietic progenitor cells to T cells.

For your convenience, this supplement is available either individually or as part of the StemSpan™ T Cell Generation Kit or the STEMdiff™ T Cell Kit. In this protocol, CD34+ cells are first differentiated to CD7+CD5+ lymphoid progenitor cells, using StemSpan™ Lymphoid Progenitor Expansion Supplement (10X), and then further directed to differentiate to DP T cells.

StemSpan™ Lymphoid Differentiation Coating Material (100X) is intended for use in combination with StemSpan™ T Cell Progenitor Maturation Supplement (10X), StemSpan™ SFEM II medium, and StemSpan™ Lymphoid Progenitor Expansion Supplement (10X).

For detailed information on protocols for expansion and differentiation of CD34+ cells with StemSpan™, please explore the Technical Manual and Educational Materials.
Subtype
Supplements
Cell Type
Hematopoietic Stem and Progenitor Cells
Species
Human
Application
Cell Culture
Brand
StemSpan
Area of Interest
Cancer, Immunology, Stem Cell Biology, Cell Therapy Development
Formulation Category
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 #
09925
Lot #
All
Language
English
Document Type
Technical Manual
Catalog #
09925
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.