Introduction |
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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).
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. 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.
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.
Collection
In the FBS manufacturing process, whole blood is collected aseptically in disposable sterile plastic bags and allowed to 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.
Handling Serum Products
Using Serum - Although the product has been sterile filtered, aseptic procedures must be followed at all times.
Granules, flocculent material or turbidity may develop after thawing. This particulate matter does not alter the performance of the serum as a supplement for cell culture medium. Repeated freezing/thawing of serum may increase the amount of precipitate and is therefore not recommended. If you do not intend to use an entire bottle of serum aliquot it into usable quantities in sterile containers before freezing a second time.
Wipe the outside of each bottle with a suitable disinfectant solution, before setting it on the work surface. Remove the heat seal and wipe the outside of the cap with the disinfectant solution.
All serum products should be treated as potentially harmful and appropriate
care should be taken when handling them.
Raw Materials
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.
ProcessingSelected 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. Biological Industries processes FBS through a sequence of prefilters
and membrane filters. The 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.
Serum products are produced in a controlled environment (clean rooms)
designed to carefully control air pressure and particulate matter.
The manufacturing area is a class 100,000 (ISO 8) environment. The sterile
bottles and equipment are stored in a class 10,000 (ISO 7) environment, and
the filling room is a class 1000 (ISO 6) environment with class 100 (ISO 5)
laminar air flow sterile bench.
Clean rooms are monitored on a regular basis for particulate and microbial
levels to ensure that the air handling system, cleaning protocols and
personnel maintain standards control.
After filling, the final product is quickly frozen to -20ºC and held in quarantine until all
quality control tests have been completed.
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Quality Control |
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.
Physical and chemical tests
Electrophoretic Pattern
pH
Osmolality
Total proteins
Albumin
IgG
Hemoglobin
Globulins
Biochemicals Tests
The following tests are conducted on each lot of FBS:
Alanine Transaminase (ALT)
Alkaline Phosphatase
Aspartate Aminotransferase (ast)
Bilirubin - total
Bilirubin - direct
Calcium
Chloride
Cholesterol
Creatinine
Creatinine Kinase (CK)
Gamma-Glutamyl Transferase (GGT)
Glucose
High Density Lipoproteins (HDL)
Lactate Dehydrogenase
Low density Lipoproteins (LDL)
Phosphorous (Inorganic)
Potassium
Sodium
Triglycerides (TG)
Uric Acid
Microbiological tests
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 Diarrhea (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 standard Limulus Amebocyte Lysate (LAL)
with the kinetic turbidimetric method.
Biological performance (cell growth)
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 Vero 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. Vero
cells (ATCC, CCL 81): plating efficiency.
MRC-5 cells (ATCC, CCL 171): 3 passages test.
Hybridoma cells: cell growth.
StabilityFBS 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.
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Quality Assurance |
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) exists 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.
Biological Industries' products are manufactured in compliance with the
quality management standard ISO 9001:2000 and ISO 13485:2003.
Certifications are 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 has
been issued to Biological Industries by the European Directorate for the
Quality of Medicines (EDQM) in accordance with monographs of the
European Pharmacopeia.
All documents and certifications are available upon request. |
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Foetal Bovine Sera (FBS) |
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. |
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Biological Industries offers you the following certified sterile Foetal
Bovine Serum Products: |
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| Product Name |
Catalogue No. |
Unit Size |
Storage Temp. |
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| Certified Foetal Bovine Serum |
04-001-1A
04-001-1B |
500ml
100ml |
-20ºC
-20ºC |
Certified Foetal Bovine Serum
Heat Inactivated |
04-121-1A
04-121-1B |
500ml
100ml |
-20ºC
-20ºC |
Certified Foetal Bovine Serum Qualified
for Human Embryonic Stem Cells |
04-002-1A
04-002-1B |
500ml
100ml |
-20ºC
-20ºC |
Certified Foetal Bovine Serum Qualified
for Human Embryonic Stem Cells
Heat Inactivated |
04-222-1A
04-222-1B |
500ml
100ml |
-20ºC
-20ºC |
| Certified Foetal Bovine Serum Functionally Tested for use with Tetracycline Regulated Systems |
04-005-1A
04-005-1B |
500ml
100ml |
-20ºC
-20ºC |
| Certified Foetal Bovine Serum Functionally Tested for use with Tetracycline Regulated Systems Heat Inactivated |
04-125-1A
04-125-1B |
500ml
100ml |
-20ºC
-20ºC |
| Certified Foetal Bovine Serum Dialyzed |
04-011-1A
04-011-1B |
500ml
100ml |
-20ºC
-20ºC |
| Certified Foetal Bovine Serum Charcoal-Stripped |
04-201-1A
04-201-1B |
500ml
100ml |
-20ºC
-20ºC |
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European Grade Approved FBS: |
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| Product Name |
Catalogue No. |
Unit Size |
Storage Temp. |
|
| Foetal Bovine Serum (FBS) European Grade |
04-007-1A
04-007-1B |
500ml
100ml |
-20ºC
-20ºC |
Foetal Bovine Serum (FBS) European
Grade
Heat Inactivated |
04-121-1A
04-121-1B |
500ml
100ml |
-20ºC
-20ºC |
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Heat Inactivated FBS |
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| Product Name |
Catalogue No. |
Unit Size |
Storage Temp. |
|
Certified Foetal Bovine Serum
Heat Inactivated |
04-121-1A
04-121-1B |
500ml
100ml |
-20ºC
-20ºC |
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| 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 proteins activity. |
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