外泌体是细胞分泌的直径为30-150 纳米的细胞外囊泡，携带蛋白质、核酸（如miRNA, mRNA）和脂质等生物活性分子，在细胞间通讯中扮演关键角色。在标准胎牛血清中，富含来自牛细胞的外泌体。当使用标准FBS 进行细胞培养时，这些牛源外泌体会被实验细胞摄取，严重干扰细胞自发性外泌体的分泌谱、功能研究以及下游的组学分析（如RNA-seq、蛋白质组学）。去外泌体胎牛血清是经过特殊超速离心工艺处理，已最大程度去除其中内源性外泌体及微囊泡的胎牛血清。它能在支持细胞良好生长的同时，极大降低背景外泌体对实验结果的干扰，是外泌体
研究、细胞外囊泡功能学实验、以及需要“洁净”细胞培养环境的理想选择。
制备原理
本产品采用超速离心法，这是去除外泌体的标准方法。
1. 前处理： 选用优质、低内毒素的胎牛血清。
2. 低速离心： 去除细胞碎片和大颗粒物。
3. 超高速离心： 在100,000 - 120,000 × g 的超高离心力下，长时间离心（通常>16 小时），使血
清中的外泌体、病毒及亚细胞器沉淀至管底。
4. 无菌收集： 小心收集上清液，即为去外泌体血清。
5. 过滤除菌： 经0.1 μm 孔径滤膜过滤，确保无菌。
6. 质量控制： 通过纳米颗粒追踪分析、蛋白质印迹（检测外泌体标志物如CD63、TSG101、Flotillin-1的缺失）等方法验证外泌体去除效率（通常去除率>95%）。
产品特性与质量控制
外泌体去除效率：> 95% （经NTA 验证）
外泌体标志物： 经WB 检测，CD9/CD63/TSG101 等标志物信号极低或检测不到。
生物活性： 保留血清中基本的生长因子、激素、营养物质，支持多种细胞系的正常增殖与活力
无菌性： 终端0.1 μm 过滤，确保无细菌、支原体、真菌污染。
内毒素水平：≤10 EU/mL
病毒筛查： 牛腹泻病毒、牛腺病毒、牛细小病毒、呼肠孤病毒3 型、狂犬病毒、牛副流感病毒3 型，以上病毒荧光抗体检测均应为阴性，应无细胞病变，血吸附应为阴性。
应用领域
1. 外泌体/细胞外囊泡研究：
细胞培养： 用于培养分泌外泌体的细胞，确保收集到的外泌体来源于实验细胞，而非血清背景。
功能实验： 研究细胞自发性分泌的外泌体在受体细胞中的生物学功能（如增殖、迁移、分化、信号传导）。
2. 条件培养基制备： 用于制备“洁净”的条件培养基，进行无外泌体干扰的旁分泌效应研究。
3. “组学”研究： 对外泌体进行蛋白质组学、转录组学、代谢组学分析时，使用本产品可避免牛源分子污染，提高数据准确性和可信度。
4. 病毒模拟物研究： 降低血清囊泡对病毒样颗粒或病毒研究的背景干扰。
5. 对囊泡摄取敏感的实验： 如干细胞共培养、肿瘤微环境研究等。
使用指南
1. 解冻与保存：
保存： 建议于-20℃ 或-80℃长期保存，避免反复冻融。可分装后冻存。
解冻： 置于2-8℃冰箱缓慢解冻过夜，或在水浴中于37℃快速解冻。解冻后请轻轻混匀。
使用后保存： 解冻后未用完的血清，应尽快放回-20℃。建议在2-4 周内使用完毕。
2. 使用方法：
与常规FBS 使用方法相同，可按所需比例（通常为5%-20%）直接添加到基础培养基（如DMEM,RPMI-1640）中。
重要提示： 由于超速离心过程可能损失了部分不稳定的生长因子，部分敏感细胞或原代细胞的生长速度可能略慢于使用标准FBS 时。建议进行细胞生长曲线测试以优化培养条件。对于极其敏感或难养的细胞，可考虑将本产品与标准FBS 以一定比例混合使用。
3. 实验对照设置：
阴性对照： 使用本产品培养的细胞及其分泌的外泌体。
阳性对照/基准对照： 可使用标准FBS 培养的同一细胞系，以评估去外泌体处理对细胞表型及外泌体产出的影响。
注意事项
1. 并非完全无外泌体： “去外泌体”是一个相对概念，是指外泌体含量被极大地降低，而非100%绝对去除。
2. 细胞适应性： 首次使用时，强烈建议监测细胞的形态、增殖率和基本功能，以确认细胞的适应性。
3. 避免污染： 操作时使用无菌技术。分装和使用过程避免引入外源性外泌体污染。
4. 配套试剂： 用于收集外泌体的条件培养基，也建议使用基于本产品配制的培养基。
常见问题解答
Q1: 去外泌体血清可以完全替代标准FBS 吗？
A1:对于大部分常规细胞培养（非外泌体研究），标准FBS 是更经济高效的选择。去外泌体血清是专门为外泌体及相关研究设计的特殊工具。
Q2: 如何自行验证其去外泌体效果？
A2:可取少量本品与等量标准FBS，通过相同流程（如超速离心）提取外泌体，然后进行：
NTA：比较颗粒浓度。
WB：检测外泌体标志蛋白（如CD63）的含量。
电镜：观察囊泡形态和数量。
Q3: 使用后细胞状态变差怎么办？
A3:首先检查是否无菌操作。如果确认是血清适应性问题，可尝试：
降低血清使用比例。
将本品与少量标准FBS（如1%-2%）混合使用。
延长细胞适应时间（传代2-3 次后通常能稳定）。
本产品仅用于科研或工业用途，不适用于临床治疗。
用户需根据自身实验体系验证血清适用性，生产方不承担因操作不当导致的实验结果偏差责任。
Exosomes are extracellular vesicles secreted by cells, with diameters ranging from 30 to 150 nanometers. They carry bioactive molecules such as proteins, nucleic acids (e.g., miRNA, mRNA), and lipids, and play a key role in intercellular communication. Standard fetal bovine serum (FBS) is rich in bovine‑derived exosomes. When using standard FBS for cell culture, these bovine exosomes can be taken up by experimental cells, seriously interfering with the secretion profile of endogenous exosomes, functional studies, and downstream omics analyses (such as RNA‑seq and proteomics). Exosome‑depleted FBS is produced through a specialized ultracentrifugation process to maximize the removal of endogenous exosomes and microvesicles. It supports robust cell growth while greatly reducing background exosome interference, making it an ideal choice for exosome research, extracellular vesicle functional studies, and experiments requiring a “clean” cell culture environment.
Preparation Principle
This product employs ultracentrifugation, the standard method for exosome removal.
1.Pretreatment: Select high‑quality, low‑endotoxin fetal bovine serum.
2.Low‑speed centrifugation: Remove cell debris and large particles.
3.High‑speed ultracentrifugation: Centrifuge at 100,000–120,000 × g for an extended period (typically >16 hours) to pellet exosomes, viruses, and subcellular organelles.
4.Aseptic collection: Carefully collect the supernatant, which is the exosome‑depleted serum.
5.Filter sterilization: Filter through a 0.1 μm pore‑size membrane to ensure sterility.
6.Quality control:Validate exosome removal efficiency (typically >95%) using Nanoparticle Tracking Analysis (NTA) and Western blot (to confirm the absence of exosome markers such as CD63, TSG101, and Flotillin‑1).
Product Characteristics and Quality Control
Exosome removal efficiency: >95% (verified by NTA)
Exosome markers: By Western blot, signals for CD9, CD63, TSG101, etc. are extremely low or undetectable.
Bioactivity: Retains essential growth factors, hormones, and nutrients in the serum; supports normal proliferation and viability of various cell lines.
Sterility: Terminal 0.1 μm filtration ensures freedom from bacterial, mycoplasma, and fungal contamination.
Endotoxin level: ≤10 EU/mL
Virus screening: Fluorescent antibody tests for bovine diarrhea virus, bovine adenovirus, bovine parvovirus, reovirus type 3, rabies virus, and bovine parainfluenza virus type 3 should all be negative; no cytopathic effect should be observed, and hemadsorption should be negative.
Applications
1.Exosome / extracellular vesicle research
Cell culture: For culturing cells that secrete exosomes, ensuring that collected exosomes originate from the experimental cells rather than from serum background.
Functional studies: To investigate the biological functions (e.g., proliferation, migration, differentiation, signal transduction) of endogenously secreted exosomes on recipient cells.
2.Conditioned medium preparation: For preparing “clean” conditioned media to study paracrine effects without exosome interference.
3.“Omics” studies: When performing proteomic, transcriptomic, or metabolomic analyses of exosomes, use this product to avoid bovine‑derived molecular contamination, thereby improving data accuracy and reliability.
4.Viral mimic studies: To reduce background interference from serum vesicles in studies on virus‑like particles or viruses.
5.Experiments sensitive to vesicle uptake: Such as stem cell co‑culture, tumor microenvironment studies, etc.
Usage Guidelines
1.Thawing and storage
Storage: Recommended for long‑term storage at –20°C or –80°C; avoid repeated freeze‑thaw cycles. Aliquoting before freezing is advised.
Thawing: Thaw slowly at 2–8°C overnight or rapidly in a 37°C water bath. After thawing, mix gently.
Post‑use storage: After thawing, unused serum should be returned to –20°C as soon as possible. It is recommended to use within 2–4 weeks.
2.Usage method
Use in the same manner as conventional FBS. Add directly to basal media (such as DMEM, RPMI‑1640) at the desired ratio (typically 5%–20%).
Important note: Due to the ultracentrifugation process, some unstable growth factors may be lost; therefore, the growth of certain sensitive or primary cells may be slightly slower than with standard FBS. It is advisable to perform cell growth curve tests to optimize culture conditions. For extremely sensitive or difficult‑to‑culture cells, consider mixing this product with standard FBS at a certain ratio.
3.Experimental control setup
Negative control: Cells cultured with this product and their secreted exosomes.
Positive/benchmark control: The same cell line cultured with standard FBS, to assess the impact of exosome depletion on cell phenotype and exosome production.
Precautions
Not completely exosome‑free: “Exosome‑depleted” is a relative term, indicating that exosome content is greatly reduced, not 100% absolutely removed.
Cell adaptation: When using for the first time, it is strongly recommended to monitor cell morphology, proliferation rate, and basic functions to confirm cell adaptability.
Avoid contamination: Use aseptic techniques during handling. Avoid introducing exogenous exosome contamination during aliquoting and use.
Matching reagents: Conditioned media used for collecting exosomes should also be prepared using media based on this product.
Frequently Asked Questions (FAQ)
Q1: Can exosome‑depleted serum completely replace standard FBS?
A1: For most routine cell culture (non‑exosome research), standard FBS is a more cost‑effective choice. Exosome‑depleted serum is a specialized tool designed specifically for exosome and related studies.
Q2: How can I verify the exosome‑depletion effect myself?
A2: Take a small amount of this product and an equal amount of standard FBS, extract exosomes using the same procedure (e.g., ultracentrifugation), and then perform:
NTA: Compare particle concentrations.
Western blot: Detect exosome marker proteins (e.g., CD63) content.
Electron microscopy: Observe vesicle morphology and quantity.
Q3: What should I do if cell condition deteriorates after use?
A3: First, check for aseptic technique. If it is confirmed to be a serum adaptation issue, you may try:
Reducing the serum proportion.
Mixing this product with a small amount of standard FBS (e.g., 1%–2%).
Extending the cell adaptation period (typically stabilizes after 2–3 passages).
Intended Use and Disclaimer
This product is intended for research or industrial use only; it is not indicated for clinical treatment. Users are responsible for validating the suitability of the serum in their own experimental systems. The manufacturer assumes no liability for deviations in experimental results arising from improper handling or operation.
ZJ26F0938980
危险品化学品经营许可证（带存储）