TeSR™-AOF

cGMP 级、无动物源、稳定、无饲养层的人胚胎干细胞（ES）和诱导多能干细胞（iPS）维持培养基

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产品号 #（选择产品）

产品号 #100-0401_C

cGMP 级、无动物源、稳定、无饲养层的人胚胎干细胞（ES）和诱导多能干细胞（iPS）维持培养基

产品优势

无动物源：通过选择不含动物原料（直至二级生产过程）的培养基，降低辅助材料使用风险
稳定的细胞生长表现：支持高质量的克隆形态、牢固的附着力以及细胞扩增能力
成分稳定：包含稳定的成分（如 FGF2），在确保细胞质量的同时允许灵活的换液方案

产品组分包括

TeSR™-AOF 基础培养基，475 mL
TeSR™-AOF 20X 补充剂，25 mL

总览

在人多能干细胞（hPSC）来源的细胞治疗开发过程中，使用 TeSR™-AOF 可降低风险并获得更多高质量细胞。该产品依照相关的 cGMP 标准生产，且在二级制造过程中未使用任何动物或人源材料，因此相比仅在一级制造过程中无动物源的培养基，它具有更直接的可追溯性和更高的病毒安全性。

TeSR™-AOF 可用于稳定培养病毒安全性高、质量优良的多能干细胞（PSCs），并可根据您的时间安排灵活换液方案，适用于您选择的任何细胞系。为提升细胞质量，尤其是在限制性换液方案的情况下，我们对培养基中的关键成分（如成纤维细胞生长因子 2，FGF2，又称碱性 FGF 或 bFGF）进行了稳定处理。因此，TeSR™-AOF 能在维持细胞质量和性能的同时，支持每天换液或限制性换液的培养方案。

TeSR™-AOF 与多种培养基质兼容，包括 Corning® Matrigel® hESC 合格基质（康宁目录号 #354277）、Vitronectin XF™（目录号 #07180）和CellAdhere™ Laminin-521（目录号 #77003）。

该培养基及其成分的生产过程中，至少在二级生产阶段，均未使用任何动物或人体来源的材料。用于制备 TeSR™-AOF 完整培养基的每一批次 TeSR™-AOF 20X 补充均已经过了人多能干细胞的培养测试以验证性能。

TeSR™-AOF 按照相关的 cGMP 生产，确保最高的质量和一致性，从而获得可重复的结果。了解更多关于STEMCELL 的质量和合规性信息。

如需申请 TeSR™-AOF 的 FDA 主文件授权书 (LOA)，请点击此处

实验数据

Figure 1A. hPSCs Maintained in TeSR™-AOF with Daily and Restricted Feed Schedules Exhibit Comparable Colony Morphology

hPSCs were maintained on Vitronectin XF™ for five passages. Phase-contrast images were taken on day 7 after seeding. For restricted feeds, hPSCs were fed with a double volume (4 mL) of medium on day 2 after passage, followed by two consecutive skipped days of feeds, with a final single-volume feed (2 mL) on day 5, prior to passaging on day 6 or 7.

Figure 1B. hPSCs Maintained in TeSR™-AOF with Daily and Restricted Feed Schedules have Comparable Expansion Rates

hPSCs were maintained on Vitronectin XF™ for five passages. At the end of each passage cell counts were obtained using the Nucleocounter®️ NC-200 ChemoMetec automated cell counter to count DAPI-stained nuclei. The log2 transformed cumulative fold expansion was plotted against time in culture (days).

Figure 2. Native bFGF Levels are Stabilized at 37°C in TeSR™-AOF

TeSR™-AOF and TeSR™-E8™ were incubated at 37°C for 24, 48, and 72 hours. FGF2 levels were measured by Meso Scale Discovery (MSD) immunoassay; data was normalized to t = 0 levels for TeSR™-E8™ and TeSR™-AOF, respectively. FGF2 levels in TeSR™-AOF remain at 36.7 ± 5.61% of t = 0 levels at 72 hours when incubated at 37°C. Data representative of n = 3 biological replicates ± SD.

Figure 3. hPSCs Cultured in TeSR™-AOF with Restricted Feeding Maintain Demonstrate Classic hPSC Colony Morphology

hPSCs maintained in TeSR™-AOF were passaged as aggregates with ReLeSR™ passaging reagent every 6-7 days for greater than 10 passages. hPSCs maintained in TeSR™-AOF exhibit hPSC-like morphology, forming densely packed, round colonies with smooth edge morphology. Homogeneous cell morphology characteristic of hPSCs are observed, including large nucleoli and scant cytoplasm.

Figure 4. hPSCs Maintained in TeSR™-AOF Have Improved Attachment and Higher Overall Expansion Compared to Low-Protein Medium

(A) hPSCs cultured in TeSR™-AOF demonstrate a higher plating efficiency compared to hPSCs maintained in low-protein medium (TeSR™-E8™). Plating efficiency is calculated by seeding a known number of aggregates and comparing to the number of established colonies on day 7. (B) hPSCs maintained in TeSR™-AOF exhibit a higher average fold expansion per passage compared to TeSR™-E8™. (C) hPSCs cultured in TeSR™-AOF demonstrate consistent expansion and minimal cell line-to-cell-line variability between ES and iPS cell lines assessed. Cumulative fold expansion was measured from passage 1 to 5. Data represented as mean plating efficiency or fold expansion across 10 passages ± SD. MG = Matrigel®; VN = Vitronectin XF™.

Figure 5. hPSCs Cultured in TeSR™-AOF with Restricted Feeding Maintain a Normal Karyotype

ES (H9 & H1) and iPS (STiPS-M001 & STiPS-F016) cell lines cultured in TeSR™-AOF were screened for chromosomal abnormalities using the hPSC Genetic Analysis Kit and by G-banding at ≥ 10 passages. Representative data are shown for (A) H1 ES and STiPS-F016 hiPS cell cultures at passage 10; no common chromosomal abnormalities were detected using the hPSC Genetic Analysis Kit, and (B) H1 ES cultures; these displayed a normal karyotype by G-banding at passage 11 and 13 respectively.

Figure 6. hPSCs Cultured in TeSR™-AOF Express Markers of the Undifferentiated State and Differentiate to the Three Germ Layers

(A) hPSCs maintained in TeSR™-AOF exhibit high levels of TRA-1-60 and OCT4 by flow cytometry at passage 5 and 10. Across n = 5 cell lines, the average TRA-1-60 expression was 92.8 ± 3.77 %, and percent OCT-4 positive cells were 98.1 ± 1.79%. Data shown represent an average of passage 5 and 10 flow results for each cell line. MG = Matrigel®; VN = Vitronectin XF™. (B) Efficient differentiation to the three germ layers was demonstrated in one hES and one hiPS cell line maintained for > 5 passages in TeSR™-AOF. Cultures were processed for flow cytometry and assessed for CXCR4+/SOX17+ cells on day 5 following differentiation using the STEMdiff™ Definitive Endoderm Kit. Cultures were processed for flow cytometry and assessed for Brachyury (T)+/OCT4- cells on day 5 following differentiation in STEMdiff™ Mesoderm Induction Medium. Cultures were processed for flow cytometry and assessed for PAX6+/Nestin+ cells on day 7 following monolayer differentiation using STEMdiff™ Neural Induction Medium.

Figure 7. hPSCs Culture in TeSR™-AOF Differentiate to Cardiomyocytes with the STEMdiff™ Cardiomyocyte Differentiation Kit

Efficient differentiation to cardiomyocytes was demonstrated in 1 hES and 1 hiPS cell line maintained in TeSR™-AOF using the STEMdiff™ Cardiomyocyte Differentiation Kit. Expression of cardiac troponin T (cTnT) was assessed by immunocytochemistry (ICC) and by flow cytometry.

产品说明书及文档

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Product Information Sheet 1

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TeSR™-AOF

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