Download Biological Industries Israel Beit Haemek Ltd. Product

Biological Industries Israel
Beit Haemek Ltd.
Version: 1.2
Date: 11/2009
Pages: 1 of 3
Product Profile
Product Name:
Peripheral Blood Karyotyping Medium, with
Phytohemagglutinin (PHA-M)
01-201-1
(A)500ml ;(B)100ml
Frozen Brown-Colored Solution
-20°C
Please Refer to the Product Label
Product Catalog Number
Unit Size Availability:
Formulation:
Specified Storage Conditions:
Stability: (Under Specified Handling &
Storage)
Important Note! Please read the MSDS and Product Profile carefully in their entirety before using this material for possible safety
precautions and potential hazards.
Product Description:
Peripheral Blood Karyotyping (PBK) Medium with Phytohemagglutinin (PHA-M) has been specifically optimized for short-term culture of
peripheral blood lymphocytes for chromosome evaluation. Lymphocytes do not normally undergo subsequent cell divisions. However, in the
presence of a mitogen, lymphocytes are stimulated to undergo mitosis by DNA replication. After 48-72 hours, a mitotic inhibitor is added to stop
mitosis in the metaphase stage. After treatment utilizing a hypotonic solution, fixation and staining, the chromosomes can then be observed
and evaluated microscopically. Routine hematological screening by means of cytogenetic analysis of human lymphocytes is an important tool
in clinical diagnosis and cancer therapy. PBKaryotyping Medium is based on RPMI-1640 basal medium supplemented with L-Glutamine,
antibiotics (Gentamicin) and with Phytohemagglutinin (PHA-M). Each batch of Karyotyping Medium is tested in a certified cytogenetics
laboratory for Chromosome Analysis Performance (CAP).
Karyotyping
Blood cell karyotyping is an important tool in modern Human Cytogenetics. Karyotyping is just one of the numerous techniques that allow us to
investigate several thousands of possible genetic developmental abnormalities, chromosomal defects, their frequency in the population at-large
and the relationship between specific chromosomal abnormalities and phenotypic effects.
Human cytogenetic studies entail the examination of a stimulated lymphocyte after blocking cell division at metaphase with an inhibitor of the
mitotic spindle formation. The nuclear membrane breaks down and chromosome condensation takes place as usual, except that the
chromosomes fail to organize themselves into a metaphase plate. In certain types of cells, the chromosomes do not line up at the metaphase
plate and instead move back and forth between the poles in a random fashion, only roughly lining up along the mid-line. This gives an
appearance unlike the natural metaphase state in that the chromosomes are free within the cytoplasm.
Staining
Subsequent processing and staining allows clear visualization of the chromosomes. The chromosomes can be stained either by a technique
that gives a fairly uniform intensity, or that presents differential staining along the length of the chromosome.
Peripheral Blood Karyotyping (PBK) Medium with Phytohemagglutinin (PHA-M) is a complete, ready-to-use proprietary blend of
ingredients. This optimal in vitro cell-culture environment conditions mimic those of the in vivo environment including basic nutritional
requirements, osmotic pressure, physiological pH, temperature among other considerations. At a minimum, it consists of the typical amino
acids, energy sources, inorganic salts and vitamins that are all part and parcel of a pre-tested complete media to assist the cells in meeting
their metabolic demands.
The PBK Medium with PHA-M with its basal medium foundation supplies the proprietary blend of nutrients, antibiotics, Heparin, FBS and LGlutamine to complement and segue into the final touch niche requirements in a ready-to-use formulation. No additions are required except the
addition of 2-4ml of PHA-M (Product Cat. No.: 12-006-1H) per 100ml of PBK Medium.
Serum and Serum Products
The medium also contains Pre-Screened and Tested Serum. Each and every Serum lot undergoes the most stringent and rigorous Quality
Control/Assurance standards and protocols testing all raw materials and finished products in order to meet the demands of international
markets and ensure high quality and consistency. All our serum products meet approved compliance validation and specifications prior to use
and/or release of the final product to the end-user. Our Fetal Bovine Serum (FBS) undergoes a methodical and comprehensive battery of
Physico-Chemical, Microbiological and Biological Performance Testing Procedures. Each batch is traceable, well-documented from source of
origin through the meticulous and systematic Quality Control process. All documentation and certification is available upon request.
It can be said that our PBK Medium without PHA-M typically represents the necessary requirements for promoting cell growth and
differentiation by providing growth-promoting factors that supply and maintain the essential levels of indispensable components that define
such specific cell-culture media. It is a one-bottle complete blend formulated for your convenience. Simply thaw and use according to your
protocols.
Biological Industries, Kibbutz Beit Haemek 25115 Israel
Telephone: 972-4-9960-595 Fax: 972-4-9968-896
Web Site: www.bioind.com
1
E-Mail: [email protected]
Biological Industries
Page 2 of 3 Pages
The PBK Medium with PHA-M contains numerous important basic constituents in a ready-to-use formulation that includes a typical and wide
variety of elements, among others:
♦
Amino Acids
♦
Glucose
♦
Inorganic Salts
♦
Lipids
♦
Phenol Red
♦
Vitamins
♦
Trace Elements
Some Predominant Characteristics of PBK Medium with PHA-M includes:
§
Ready-To-Use, All-In-One, Pre-Tested Formulation
§
Serum-Supplemention and thereby eliminates Serum-Screening Down-Time
§
Chromosome Karyotyping Turn-Around Time is reduced
§
More Uniform & Consistent Media Performance
§
Sterile-Filtered(0.1µ),Cell-Culture and Endotoxin-Tested
§
Mitotic Index Tested
PBK Medium with PHA-M offers numerous applications. Some of which include:
1)
The Utility of Human Lymphocyte Cell Culture
2)
The Ease of Chromosomal Anomaly Visualization
3)
The Simplicity of Processing and Staining
Storage, Handling, Stability Precautions and Disclaimer:
For in vitro diagnostic use only. It is not intended for therapeutic use.
PBK Medium with PHA-M should be kept frozen under defined conditions of -20°C. Prior to use, thaw the medium to refrigerator
temperatures between 2-8°C and gently swirl for a homogeneous and uniform mixture. For prolonged storage, aliquot and re-freeze. Avoid
multiple freeze/thaw cycles as this will cause a loss of product activity. When stored in the dark under specified conditions, the product is stable
until the expiry date. Do not use beyond the expiration date elucidated on the product label.
The PBK medium should be stored at 2-8°C for up to ten (10) days after thawing. The product is light-sensitive and therefore should not be left
in the light. When stored in the dark under ideal conditions, the product is stable until the expiry date.
As with any other liquid media formulations,deterioration of liquid media may be recognized by any of the following characteristics, among
others including: (a). color change, (b). presence of clumping/flocculent debris/ granulation/ particulates\ precipitates or sediments (c).
insolubility,(d). and/or decrease in expected performance parameters. Any material described above should not be used and therefore
discarded.
Although each batch of Karyotyping Medium is tested in a certified cytogenetics laboratory for Chromosome Analysis Performance(CAP), the
use of Biological Industries PBK Medium does not guarantee the successful outcome of any Chromosome Analysis Testing.
Instructions/Procedure:
1)
2)
3)
Take a bottle of Peripheral Blood Karyotyping (PBK) Medium with Phytohemagglutinin (PHA-M) from specified storage at
-20°C and read the label.
Thaw the medium to refrigerator temperatures between 2-8°C or by swirling in a
37°C water-bath. Gently swirl to ensure homogeneity. Avoid multiple freeze-thaw cycles. Aliquot and re-freeze unused medium
for future use.
Using aseptic/sterile technique under a laminar-flow culture hood, work according to established protocols.
Quality Control
Test
Human Blood Karyotyping:
Endotoxins:
Mitotic Index:
Osmolality:
pH:
Sterility:
Specification
Test & Record
Test & Record
Test & Record
280-340 mOsm/kg
7.2-7.60
Negative
Biological Industries, Kibbutz Beit Haemek 25115 Israel
Telephone: 972-4-9960-595 Fax: 972-4-9968-896
Web Site: www.bioind.com
2
E-Mail: [email protected]
Biological Industries
Page 3 of 3 Pages
Auxiliary Products
Product Name
Sodium Citrate Solution (0.8%)
Dulbecco's Phosphate Buffered Saline(DPBS) without Calcium and
Magnesium
L-Glutamine Solution 29.2mg/ml in Saline
L-Alanyl-L-Glutamine Solution(A Dipeptide Substitute)
Penicillin-Streptomycin Solution,10,000 units/ml Penicillin G Sodium
Salt,10mg/ml Streptomycin
Trypsin EDTA,10X Concentrate
Fetal Bovine Serum
Fetal Bovine Serum(Qualified for Human Embryonic Stem Cells)
Adult Bovine Serum
Sterile Culture-Grade Water
Cell Dissociation Solution, Non-Enzymatic
Serum-Free Cell Freezing Medium
Colchicine Solution, 10μg/ml in DPBS
Colcemid Solution, 10μg/ml in DPBS
Potassium Chloride(KCl) Solution(0.075M)
Phytohemaglglutin-M(PHA-M), Lyophilized
Note: For a list of other Antibiotics, Serum, Reagents and
Supplements, please refer to our Product Catalog, Product Profiles,
Product Guides and Internet Site.
Catalog Number
01-934-1
02-023-1
Storage Temperature
Room Temperature(15-30°)
Room Temperature(15-30°)
03-020-1
03-022-1
03-031-1
-20°C
-20°C
-20°C
03-051-1
04-001-1
04-002-1
04-003-1
03-055-1
03-071-1
05-065-1
12-003-1
12-004-1
12-005-1
12-006-1
-20°C
-20°C
-20°C
-20°C
Room Temperature(15-30°)
2-8°C
2-8°C
-20°C
-20°C
2-8°C
2-8°C
References:
1) Moorehead, P.S. et. al., "Chromosome Preparations of Leukocytes Cultured from Human Peripheral Blood," Ex. Cell Res.,
20:613-616(1960).
2) Nowell, P.C. "Phytohemagglutinin- An Initiator of Mitosis in Cultures of Normal Human Leukocytes,"
Cancer Research, 290:462-466(1960).
3) Barch, M.J.(ed.). The Association of Cytogenetic Technologists Laboratory Manual, Second Edition, (1991)
4) Biological Industries (BI )Specifications
5) Darling, D.C. and Morgan S.J. Animal Cells: Culture and Media, John Wiley & Sons, New York, 1994
6) Biological Industries (BI) Product Guide, "Human Cytogenetic Products," p.5.
Biological Industries, Kibbutz Beit Haemek 25115 Israel
Telephone: 972-4-9960-595 Fax: 972-4-9968-896
Web Site: www.bioind.com
3
E-Mail: [email protected]