FOETAL BOVINE SERUM PRODUCTS 胎牛血清产品

1. FOETAL BOVINE SERUM PRODUCTS 胎牛血清产品

2. INTRODUCTION介绍

3. Serum is commonly used as a supplement to basal growth medium in cell culture. The most common type of serum used for cell growth is foetal bovine serum (FBS), also known as foetal calf serum (FCS). 血清通常在细胞培养中作为基础生长培养基的添加剂。最常见血清是 胎牛血清（FBS），也称FCS

4. Foetal bovine serum is obtained from foetuses harvested in abattoirs from healthy dams fit for human consumption. Occasionally, there may be use of other bovine sera, such as newborn calf serum or donor bovine serum. 胎牛血清是从适合人类卫生习惯的农场收集的。有时，也可能用其他的牛血清，如新生牛血清或者供体牛血清。

5. In cell culture, serum provides a wide variety of macromolecular proteins, low molecular weight nutrients, carrier proteins for water – insoluble components, and other compounds necessary for in vitro growth of cells, such as hormones and attachment factors. Serum also adds buffering capacity to the medium and binds or neutralizes toxic components. Attempts to replace serum entirely with serum-free medium have met only with limited success. 在细胞培养中，血清供应了丰富的高分子蛋白，低分子量营养物，疏 水载体蛋白和其他为体外细胞生长必要成分如激素和贴附因子。同 时，血清可以增加培养基的缓冲能力含或者中和有毒成分。只有少量 的无血清培养基成功替代了含血清培养。

6. The selection of a serum supplement for cell culture applications is primarily dependent on the chemical definition of the basal medium, the type of cell to be grown, and the culture system being employed. 细胞培养应用中是否选择添加血清主要根据基础培养基化学成分，细 胞培养类型和培养体系来选择。

8. COLLECTION采集

9. In the FBS manufacturing process, whole blood is collected aseptically in disposable sterile plastic bags andallowedto clot. Once the serum has been separated from the clot, it is pooled and frozen. Controlling the initial collection of foetal blood is a crucial factor in the quality of the final serum product. Only raw material that meets our specifications is approved for production. 在胎牛血清制造过程，全血使用无菌一次性塑料袋收集，并凝结后分离血清，收集 和冷冻储存。控制胎牛血清的初始收集是控制最终血清产品质量的关键因素。只有 初始原料符合我们的规范时才能允许生产。

10. RAW MATERIALS原料

11. Two distinct grades of FBS are available on the world market: USDA-Grade FBS and European-Grade FBS.USDA-Grade FBS is produced from raw materials originating only from countries certified to be free of both BSE (Bovine Spongiform Encephalopathy) and FMD (Foot and Mouth Disease). This product can be freely imported into any country, and is the product of choice in all countries for manufacturing purposes. Furthermore only the use of this product allows researchers to send their cells, or the products of their cells, to collaborators in other countries with strict import regulations. All FBS processed in the Biological Industries plant is USDA-Grade. 在全球市场存在两个胎牛血清标准:美国农业部-标准（USDA）和欧洲标准。 美国农业部标准，原始血清采自未发生疯牛病（BSE）和口蹄疫（FMD）疫情的国家。可自 由进口到任何国家，可安全用于生产。而且只有使用符合这种标准血清的研究者，才被允许 将其细胞和细胞产品送至其他国家，进行合作研究。这些国家也都有严格的进口条款。所有 Biological Industries工厂加工的胎牛血清均符合USDA标准。

12. PROCESSING处理过程

13. Selected batches of serum raw material are thawed, tested for endotoxins and hemoglobin content and only the accepted material is pooled. The pooled raw material is thoroughly blended under refrigerated conditions and membrane filtered for sterility according to a well validated filtration protocol. 优选数个批次冻存血清进行解冻，检验其内毒素和血红素的含量，满足标准的才能进一 步加工冷藏条件下充分混合解冻血清，进行有效地膜过滤除菌。

14. Biological Industries processes FBS through a sequence of pre-filters and membrane filters. The final filtration step includes the use of three 0.1 micron sterilizing grade membrane filters in series. After filtration, the serum is dispensed into bottles by an aseptic filling process which has been validated to insure sterility of the final product. Biological Industries依次通过预过滤和膜过滤处理胎牛血清。最后的过滤步骤 为连续通过三个0.1微米的滤器。过滤结束，血清采用无菌工艺进行分装, 整个过程有效确保最终产品无菌。

15. The serum products are filled in a class 100 environment. All airflow on the filling bench is filtered through HEPA filters. The room is maintained under positive pressure to its surroundings. The filling environment is routinely monitored for total particle counts, viable particle counts and pressure. After filling, the final product is quickly frozen to –20oC and held in quarantine until all quality control tests have been completed. 血清生产的洁净级别为100级。所有分装工作台的都要通过高效微粒空气过滤器过 滤，并保持正压分装环境要对总微粒、悬浮微粒数和压力进行监控。 分装之后，终产品迅速-20度冻存，并隔离，直到所有的质量控制检测完成。

16. FBS QUALITY CONTROLFBS质量控制

17. Each lot of FBS is tested to confirm that the serum meets the written specifications.Final product release is done after reviewing all production and quality control records to determine compliance with all established, approved written procedures. 每批胎牛血清检测确认符合产品描述。 在确认所有产品生产和质量控制记录后发布出厂许可，以确保符合所有已确立的规程 和程序。 Physical and chemical tests: PH，Osmolality， BiochemicalProfile：Totalproteins，Albumin，Globulins， IgG，Hemoglobin 物理和化学检验： PH/ 渗透性/总蛋白/清蛋白/IgG/血红素

18. Microbiological tests微生物检测 Sterility tests: Bacterial and fungal sterility tests according to the current USP Mycoplasma contamination: according to the Code of Federal Regulations (CFR), title 9, part 113 (culture method) Viral contaminants: according to the protocols described in CFR, title 9, part 113 for Bovine Viral Diarrea (BVD), Infectious Bovine Rhinotracheitis (IBR) and Parainfluenza type 3 (PI3). Viral antibodies: FBS is screened to determine the titer of neutralizing antibodies to BVD, IBR and PI3 Endotoxins: the test is performed using the standarLimulus Amebocyte Lysate (LAL) with the kineticturbidimetric method. 无菌检测： 根据常规的USP进行细菌和真菌无菌检验。 支原体污染：依照CFR培养方法检测. 病毒污染：根据CFR标准程序，检测胎牛病毒痢疾（BVD），牛传染性鼻气管炎病（IBR） 和牛副流行性感冒﹝PI3）。 病毒抗体：筛选FBS以确定BVD，IBR和PI3中和抗体滴度。 内毒素： 检验使用标准的LAL试剂用比浊法进行检测。

19. Biological performance (cell growth)生物特性 The cell growth tests are designed to check the efficacy of the FBS in promoting cell growth. Cells used are fibroblasts (MRC-5 diploid normal cells), epithelial cells (Hep-2) and hybridoma cells. Each test is conducted using the tested serum and a validated control lot. Growth promotion using MRC-5 cells is evaluated through several subculture generations to observe any evidence of cytotoxicity and morphological changes of the cells. Hep-2 cells (ATCC, CCL 23): growth curve MRC-5 cells (ATCC, CCL 171): 3 passages test Hybridoma cells: cloning efficiency test 细胞生长检测设计为核实促进细胞生长的胎牛血清的功效。使用的细胞是成纤维细胞（MRC-5双倍染色体正常细胞），上皮细胞（Hep-2）和杂交瘤细胞。每次测试用检测血清和一个有效的控制批号。使用MRC-5细胞，通过三个以上结构重生生长促进。 Hep-2细胞（ATCC，CCL 23）：生长曲线 MRC-5细胞（ATCC，CCL171）：3阶段测试。 杂交瘤细胞：克隆效率检测。

20. STABILITY稳定性

21. FBS stability at –20oC temperature was evaluated with several cell types for long periods. The FBS did not lose its performance for 55 months (4.5 years) with all the cells tested. Storage of FBS at –20oC without defrosting will maintain the quality of the FBS at least until the expiration date stated on the label. 胎牛血清在-20度保存的稳定性的评估，通过数个不同细胞系的长期培养进行。培养表现 可以保持55个月（4.5年）不变。不解冻的话，-20度储存的胎牛血清至少到标签注明的 日期仍能保持培养表现。

22. QUALITY ASSURANCE质量保证

23. The FBS production process is carried out under controlled conditions in a controlled environment. The steam-in-place (SIP) sterilization, filtration for sterility and filling are validated as required for key aseptic processes. A dossier (Device Master record) exist for serum with all relevant data concerning serum production. The production process from the raw material to the final product in storage, as well as the quality control tests and results, are documented and filed to ensure traceability and control of the process. 胎牛血清生产过程在一个控制环境中可控条件进行。SIP灭菌，过滤除菌和分装被做为关键 生产步骤，可以保证效果。与血清生产相关的数据作为专门的档案记录(Device Master record）。从未加工原料到最后产品储存整个生产过程，以及质量控制检测和结果都有备 案，并且确保追朔和过程控制。

24. Biological Industries products are manufactured in compliance with the quality management standard ISO 9001:2000 and certification is available upon request. In addition, the FBS production process conforms to the In Vitro Diagnostics Directive (IVDD 78/79/EC) of the European Parliament. Therefore, our FBS received the CE mark making it eligible for sale in the European Union for in vitro diagnostics. A Bovine Spongiform Encephalopathy (BSE) certificate of suitability had been issued to Biological Industries by the European Directorate for the Quality of Medicines (EDQM) via the process of certification of suitability of monographs of the European Pharmacopoeia. All documents and certifications are available upon request. Biological Industries 产品制造符合质量管理标准ISO9001：2000，并可提供品质证明。 另外，胎牛血清生产过程遵循欧洲国会关于体内诊断指示（IVDD78/79/EC）的标准。因此，我们的胎牛血清有CE标记，可以在所有欧洲国家合法出售，用于体外诊断。 欧洲药品质量管理局为Biological Industries颁发了BSE安全许可证书，其产品通过了欧洲药典的检测标准。可以提供这方面的文件和证明。

25. SPECIAL PROGRAMS AND FBS PRODUCTS(Prepared from USDA-grade FBS)特殊加工工艺胎牛血清产品

26. SERUM RESERVATION血清预定

27. Serum, as a biological material, represents an undefined mixture of components in which composition varies from one lot to the other. Some cell types are sensitive to the variations in serum performance. Customers are encouraged to evaluate serum samples with their own culture system and cells while we reserve the quantities of the specific lots until customer testing is completed. In this way, the customer may choose the best serum for his own applications. 血清，作为一种生物原料，组分的比例在每个批次都有不同，许多细胞类型对不同批次 血清很敏感。建议客户用自己的培养体系和细胞对不同批次血清样品做出评估，我们预 留特定批次的样品，直到顾客完成检测。用这种方法顾客就会按自己的要求选出最佳的血清。

28. Gamma-irradiated FBSγ-照射 FBS Gamma irradiation is a process which involves exposing material to high energy gamma photons released by radioisotopes, such as cobalt 60. This energy is transferred from the photons to the products, and is responsible for organism inactivation by ionization of nucleic acid bonds. The irradiation of serum is intended to provide complete assurance of viral inactivation. An extensive validation study has been performed to validate the irradiation process using FBS spiked with several viruses . We have demonstrated that properties and cell culture performance of FBS are not altered by gamma irradiation exposure up to 3.5 MRad. γ-照射是将原料暴露在放射释放高能量的γ中子下，如C60。这种能量从中子转移到产品，导致通过核酸结合离子化的机体无活性。血清进行照射后可以完全保证无病毒的存在。对携带病毒的胎牛血清进行照射，研究血清的培养活性可以证明，最高至3.5Mra的γ-照射，FBS的特性和细胞培养不会改变。

29. Dialyzed FBS透析FBS Most cells grown in culture require a serum component of the growth medium to maintain their proliferative capacity. While whole serum is permissible for routine purposes, studies involving nutritional parameters or incorporation of labeled material require that the constituent under study be removed from the serum. The most commonly used method for removal of these constituents is dialysis of whole serum. For dialysis by diafiltration, serum is circulated through a hollow-fiber by the concentration method. The filtrate however, is replaced by the addition of physiological saline to the serum. 大多数细胞培养需要添加血清来维持细胞的增殖能力。尽管血清可以满足一般细胞培养需求，但在进行营养成分优化和标记特定成分进行研究的实验中，仍必须去处这些特定的成分。去除最常用的方法是透析。透析的方法是将浓缩的血清循环通过中空纤维。但是，这种透析需要在血清中添加生理盐水补充。

30. Heat Inactivated FBS热灭活 FBS Heat inactivation of serum is performed by raising the temperature of the serum to 56ºC and maintaining that temperature for 30 minutes. Heat inactivation is the method of choice to destroy complement, and to ensure that the cells will not be lysed by antibody binding. 血清热灭活通过提高血清温度到56度并且保持该温度30分钟完成。热灭活可以破坏补体，确保细胞与抗体结合不会发生裂解。

31. Human Embryonic Stem (HES) cells tested FBS胚胎干细胞（ES）检测FBS Human Embryonic Stem (ES) cells are pluripotent cells derived from the inner cell mass of the blastocyst. The stem cells can be maintained in vitro for extended periods without loss of their capacity to differentiate to all cell lineages. ES cells may differentiate in vitro to a variety of cell types. 胚胎干细胞（ES）是来源于内胚层的多能性细胞。胚胎干细胞可以长时间保存 体外，当再植入胚层时不会丢失分化为其他系细胞的能力。ES细胞可以在体外分 化为神经、肌肉、上皮和造血祖细胞。

32. The most important parameter is the maintenance of the cells in undifferentiated state. The screening of the batches is done at the Technion, School of Medicine, Haifa. The cell lines are cultured with MEF’s for 4 passages during which the following parameters are measured: • hESC colony morphology • plating efficiency • background differentiation rates • ES细胞的培养，最重要的参数之一是确保细胞的未分化状态。 • 对不同批号的血清进行细胞培养来筛选。 • 细胞线用MEF’s培养4代，其间测量以下参数：hESC克隆形态，涂板板效率，分化率。

33. Charcoal-stripped FBS脱炭 FBS Charcoal-stripped FBS is used to elucidate the effects of hormones in a variety of in vitro systems. Studies include steroid- receptor binding, steroid regulation of cellular receptors, hormone secretion of various tissues and the function of thyroid hormones. The production procedure includes the use of charcoal and dextran to remove the hormones from the FBS. 脱炭胎牛血清用于体外培养验证激素的作用效果。研究包括类固醇受体连接，细胞受体类 固醇调节，各种组织激素分泌和甲状腺激素功能。生产程序为使用木炭和右旋糖苷去除胎 牛血清中的激素。

34. FOETAL BOVINE SERUM FAQS胎牛血清常见问题

35. What is the difference between Foetal Bovine Serum (FBS) and Foetal Calf Serum (FCS)? They are the same and describe exactly the same serum product. FBS和FCS的区别？ 他们是对同一种血清产品的同样确切的描述。

36. My FBS contains precipitates. What are they and what should I do? The precipitates contain fibrin (clot forming protein) and lipoproteins. This is a normal characteristic and will not affect product performance. To remove the precipitates, centrifuge the serum or simply let it settle to the bottom of the bottle and transfer the serum carefully to another sterile bottle. 我的FBS包含沉淀物，他们是什么，怎么处理？ 沉淀包含纤维蛋白和脂蛋白。这是正常特性，不会影响产品性质。要除去沉淀，离心血清或者简单的让其沉淀在瓶低部，将血清小心的转移到另一个无菌瓶里。

37. My FBS arrive partially thawed, what should I do? Let the serum defrost completely, swirl the serum bottle gently and then re-freeze the serum. The quality of the serum will not have been affected. 我的FBS部分解冻，该怎么办？ 让血清完全解冻，轻轻旋转血清瓶， 然后重新冷冻血清。血清质量不会受到影响。

38. How do I heat-inactivate serum Serum heat-inactivation is performed in a waterbath at 56ºC for 30 minutes. The water level should be higher than the level of the serum. Monitor the temperature in a reference bottle containing water at the same volume during the heat inactivation. You must swirl the bottles to mix the serum every 5 minutes during heat inactivation to insure uniform heating. Use a calibrated thermometer only! 怎样 热灭活血清？ 血清热灭活是在水浴56度，保持30分钟。水位应该高于血清水位。在热灭活过程中，温度的 调节，通过监控含同样体积的水的参照瓶里的水温进行调节。在热灭活过程必须旋转瓶子每 5分钟混合一次，确保受热均匀 使用的温度计必须经过校准！

39. I have a jellylike fraction on the bottom of the bottle. What is it? As a result of improper heat inactivation of the serum (temperature above 56ºC, more than 30 minutes), or inactivation without mixing the serum, a protein denaturation caused the jellylike fraction on the bottom of the bottle. Do not use this serum. 在瓶底有絮状的物质，他是什么？ 当血清热灭活不恰当（温度高于56度，高于30分钟），或者灭活中没有混合均匀，蛋白质变 性引起瓶底絮状沉淀。 不要使用这种血清。

40. How should I thaw FBS? We recommend thawing the serum at 2-8ºC. However, if necessary, you may thaw the serum at room temperature. Swirl the bottles gently to mix the serum during the thawing process. 如何解冻 FBS？ 我们建议2-8℃解冻血清，但是，也可以在室温解冻血清。在解冻过程轻轻晃动瓶子混合血清。

41. Why is the color of the FBS not exactly the same as with my previous lot? The color of FBS is brown to brown-red. It is dependent mainly on the hemoglobin concentration in the specific lot. The color does not affect the FBS performance. 为什么FBS的颜色与原先批号的颜色不能完全一致？ FBS的颜色是棕色到棕红色。主要依靠在特定批号中血红素浓度。颜色不会影响FBS的性能。

42. Is the FBS raw material free of BSE? FBS sold by Biological Industries has always been produced from raw materials originating from countries that are free of BSE according to the OIE (Office International des Epizootie). FBS未加工原料不含BSE吗？ Biological Industries出售的FBS原料来源于根据OIE未发生BSE疫情的国家。

43. How can I be certain regarding the country of origin of the raw material that was used to manufacture sterile FBS? Ask Biological Industries for a copy of the original raw material veterinary documents. 如何确认FBS原料来源国？ 要求Biological Industries公司提供原始的兽医文件。