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StemSpan™淋系祖细胞扩增添加物（10X）

用于人CD34+细胞扩增及分化为淋系祖细胞的添加物

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产品号 #（选择产品）

产品号 #09915_C

用于淋系祖细胞扩增与分化的添加物

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总览

实验数据

产品说明书及文档

应用领域

总览

使用StemSpan™淋系祖细胞扩增添加物（10X），可选择性促进从人脐带血（CB）样本中分离的CD34+细胞扩增并分化为CD7+CD5+ T祖细胞（pro-T细胞）。

该添加物是StemSpan™ T细胞生成试剂盒或STEMdiff™ T细胞分化试剂盒的组分之一，也可单独购买，需搭配StemSpan™ SFEM II培养基以及用StemSpan™淋系分化包板材料（100X）包被的培养板使用。如需将T祖细胞进一步成熟为CD4+CD8+双阳性（DP）T细胞，可使用StemSpan™ T细胞祖细胞成熟添加物（10X）。

关于使用StemSpan™进行CD34+细胞扩增与分化的详细实验方法，请参阅技术手册与教育资源。

分类
专用培养基，添加剂

细胞类型
造血干/祖细胞，T 细胞

种属
人

应用
细胞培养，分化，扩增

品牌
StemSpan

研究领域
癌症，细胞疗法开发，药物发现和毒性检测，免疫学，干细胞生物学

制剂类别
无血清

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实验数据

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).

产品说明书及文档

请在《产品说明书》中查找相关支持信息和使用说明，或浏览下方更多实验方案。

Document Type

Product Name

Catalog #

Lot #

Language

Document Type

Product Information Sheet

Product Name

StemSpan™ Lymphoid Progenitor Expansion Supplement (10X)

Catalog #

09915

Lot #

All

Language

English

Document Type

Technical Manual

Product Name

StemSpan™ Lymphoid Progenitor Expansion Supplement (10X)

Catalog #

09915

Lot #

All

Language

English

Document Type

Safety Data Sheet

Product Name

StemSpan™ Lymphoid Progenitor Expansion Supplement (10X)

Catalog #

09915

Lot #

All

Language

English

应用领域

本产品专为以下研究领域设计，适用于工作流程中的高亮阶段。探索这些工作流程，了解更多我们为各研究领域提供的其他配套产品。

Research Area

Workflow Stages

Workflow Stages for Hematopoietic Stem and Progenitor Cell Research

Cell Sourcing and Isolation

Expansion and Differentiation

Analysis

Workflow Stages for T Cell Research

Cell Sourcing

Cell Isolation

Cell Activation, Expansion, and Differentiation

Cell Characterization

相关材料与文献

Feeder-free differentiation of human iPSCs into natural killer cells with cytotoxic potential against malignant brain rhabdoid tumor cells Bioactive Materials 2024 Mar

Abstract

Natural killer (NK) cells are cytotoxic immune cells that can eliminate target cells without prior stimulation. Human induced pluripotent stem cells (iPSCs) provide a robust source of NK cells for safe and effective cell-based immunotherapy against aggressive cancers. In this in vitro study,a feeder-free iPSC differentiation was performed to obtain iPSC-NK cells,and distinct maturational stages of iPSC-NK were characterized. Mature cells of CD56bright CD16bright phenotype showed upregulation of CD56,CD16,and NK cell activation markers NKG2D and NKp46 upon IL-15 exposure,while exposure to aggressive atypical teratoid/rhabdoid tumor (ATRT) cell lines enhanced NKG2D and NKp46 expression. Malignant cell exposure also increased CD107a degranulation markers and stimulated IFN-? secretion in activated NK cells. CD56bright CD16bright iPSC-NK cells showed a ratio-dependent killing of ATRT cells,and the percentage lysis of CHLA-05-ATRT was higher than that of CHLA-02-ATRT. The iPSC-NK cells were also cytotoxic against other brain,kidney,and lung cancer cell lines. Further NK maturation yielded CD56?ve CD16bright cells,which lacked activation markers even after exposure to interleukins or ATRT cells - indicating diminished cytotoxicity. Generation and characterization of different NK phenotypes from iPSCs,coupled with their promising anti-tumor activity against ATRT in vitro,offer valuable insights into potential immunotherapeutic strategies for brain tumors. Graphical abstractImage 1 Highlights•Natural killer (NK) cells were derived from human induced pluripotent stem cells (iPSCs) in the absence of feeder cells.•Various maturational subtypes of iPSC-NK cells were characterized,and the phenotypic and functional properties were studied.•iPSC-NK cells of CD56bright CD16bright phenotype expressed activation markers in response to interleukin stimuli.•iPSC-NK cells were cytotoxic toward human atypical teratoid and rhabdoid tumor (ATRT) cells and other human cancer cells.•The cytotoxicity of iPSC-NK cells against various cancer cells in vitro might be translated into an in vivo immunotherapy.

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