StemSpan™ T细胞生成试剂盒

将人CD34+造血祖细胞扩增并分化为T细胞的全套试剂盒

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产品号 #（选择产品）

产品号 #09940_C

将人CD34+造血祖细胞扩增并分化为T细胞

产品组分包括

StemSpan™ SFEM II培养基，2 x 100 mL
StemSpan™淋系祖细胞扩增添加物（10X），5 mL
StemSpan™淋系祖细胞分化包板材料（100X），2 x 0.25 mL
StemSpan™T细胞祖细胞成熟添加物（10X），12.5 mL

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总览

采用两阶段的分化方法，可从起始的人脐带血（CB）CD34+细胞稳定扩增出数千个CD4+CD8+双阳性（DP）T细胞。

第一阶段中，CB CD34+细胞在含扩增添加物的培养基中培养14天，促进其增殖并分化为CD7+CD5+ T祖细胞。第二阶段中，将前14天生成的T祖细胞在含成熟添加物的培养基中继续培养4周，促使其成熟为DP细胞。该培养基试剂盒无血清且无需饲养层，消除了血清和基质细胞系引入的不稳定因素。

所有试剂盒组分可单独购买以方便您的使用。StemSpan™淋系祖细胞扩增添加物（10X）包含重组人细胞因子组合和其他添加剂，专用于选择性促进从人脐带血（CB）或骨髓（BM）样本中分离的CD34+细胞扩增及分化为CD7+CD5+ T祖细胞（pro-T）。此外，StemSpan™ T细胞祖细胞成熟添加物（10X）能进一步促使T祖细胞成熟为DP细胞，以及在进一步的刺激条件下支持DP细胞成熟为CD8单阳性（SP）T细胞。StemSpan™淋系祖细胞扩增添加物（10X）和StemSpan™ T细胞祖细胞成熟添加物（10X）需与StemSpan™ SFEM II培养基以及用StemSpan™淋系分化包板材料（100X）包被的培养板搭配使用。

可选的将DP细胞进一步成熟为CD8 SP T细胞的延长实验方法需要使用由StemSpan™ T细胞生成试剂盒所含试剂配制的完全T细胞祖细胞成熟培养基，并需要搭配使用ImmunoCult™ CD3/CD28/CD2 T细胞激活剂和IL-15。

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).

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StemSpan™ T Cell Generation Kit

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StemSpan™ T Cell Generation Kit

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09940

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09940

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StemSpan™ T Cell Generation Kit

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09940

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09940

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文献 (2)

Targeting triple-negative breast cancer using cord-blood CD34⁺ HSPC-derived mesothelin-specific CAR-NKT cells with potent antitumor activity Li et al. Journal of Hematology & Oncology 2025 Oct

Abstract

Triple-negative breast cancer (TNBC) is an aggressive subtype of breast cancer characterized by the lack of ER, PR, and HER2 expression. Its aggressive behavior, high degree of tumor heterogeneity, and immunosuppressive tumor microenvironment (TME) are associated with poor clinical outcomes, rapid disease progression, and limited therapeutic options. Although chimeric antigen receptor (CAR)-engineered T cell therapy has shown certain promise, its applicability in TNBC is hindered by antigen escape, TME-mediated suppression, and the logistical constraints of autologous cell production. In this study, we employed hematopoietic stem and progenitor cell (HSPC) gene engineering and a feeder-free HSPC differentiation culture to generate allogeneic IL-15-enhanced, mesothelin-specific CAR-engineered invariant natural killer T ( Allo15 MCAR-NKT) cells. These cells demonstrated robust and multifaceted antitumor activity against TNBC, mediated by CAR- and NK receptor-dependent cytotoxicity, as well as selective targeting of CD1d + TME immunosuppressive cells through their TCR. In both orthotopic and metastatic TNBC xenograft models, Allo15 MCAR-NKT cells demonstrated potent antitumor activity, associated with robust effector and cytotoxic phenotypes, low exhaustion, and a favorable safety profile without inducing graft-versus-host disease. Together, these results support Allo15 MCAR-NKT cells as a next-generation, off-the-shelf immunotherapy with strong therapeutic potential for TNBC, particularly in the context of metastasis, immune evasion, and treatment resistance. The online version contains supplementary material available at 10.1186/s13045-025-01736-9.

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Allogeneic CD33-directed CAR-NKT cells for the treatment of bone marrow-resident myeloid malignancies Li et al. Nature Communications 2025 Feb

Abstract

Chimeric antigen receptor (CAR)-engineered T cell therapy holds promise for treating myeloid malignancies, but challenges remain in bone marrow (BM) infiltration and targeting BM-resident malignant cells. Current autologous CAR-T therapies also face manufacturing and patient selection issues, underscoring the need for off-the-shelf products. In this study, we characterize primary patient samples and identify a unique therapeutic opportunity for CAR-engineered invariant natural killer T (CAR-NKT) cells. Using stem cell gene engineering and a clinically guided culture method, we generate allogeneic CD33-directed CAR-NKT cells with high yield, purity, and robustness. In preclinical mouse models, CAR-NKT cells exhibit strong BM homing and effectively target BM-resident malignant blast cells, including CD33-low/negative leukemia stem and progenitor cells. Furthermore, CAR-NKT cells synergize with hypomethylating agents, enhancing tumor-killing efficacy. These cells also show minimal off-tumor toxicity, reduced graft-versus-host disease and cytokine release syndrome risks, and resistance to allorejection, highlighting their substantial therapeutic potential for treating myeloid malignancies. Subject terms: Cancer therapy, Immunotherapy, Leukaemia

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物种	人类
配方	无血清

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物种

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无血清

StemSpan™ T细胞生成试剂盒

产品号 #09940

产品号 #（选择产品）

产品号 #09940_C

产品组分包括

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StemSpan™ T细胞生成试剂盒

产品号 #09940

产品号 #（选择产品）

产品号 #09940_C

产品组分包括

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