BRFF-BMZERO™ Data Sheet
Catalog No. 0401
Product Description
BRFF-BMZERO™ is a complete serum-free medium comprised of
essential amino acids, vitamins, inorganic salts, trace
elements and growth factors designed for culturing human
breast cell lines. BMZERO™ was developed and has been
extensively used for establishing new epithelial cell lines
from explants of human breast tissue and to grow immortalized
normal breast cell lines. Optimal attached monolayer cell
growth can be achieved when used in conjunction with FNC Coating Mix®.
| Product Specifications |
| Unit Size |
500 mL |
| Shipping |
Supplied frozen |
| Storage |
Store at -80°C |
| Stability |
4 - 6 weeks at 4°C, 2 years at -20°C |
| pH |
7.3 ± 0.2 |
| Osmolality |
270 - 300 mOsM |
Recommended Protocol
Most tissue cultures are initiated in serum-containing media in
order to simulate a “natural” environment thereby maximizing
cell growth. The transition from a serum-containing to a serum-free
medium generally requires an adaptive process, in which the cells
are slowly “weaned” from their dependence on serum. A sudden
transition to serum-free environment is generally not successful.
| 1. |
Cultivate the cell line to be propagated in the base medium employed.
|
| 1.1. |
If the serum concentration is greater than 5%, reduce the amount of
serum to 5%. Once the cells are growing well, proceed to step 2.
|
| 2. |
Transfer the cells to BRFF-BMZERO™ supplemented with 2% FBS (or 1:1 mixture of
base medium to BMZERO™).
|
| 2.1. |
When the cells are growing well, proceed to step 3. If the cells do not
grow well, increase the serum concentration to 3 or 4% (2:1 mixture of
base medium to BMZERO™) and repeat this step.
|
| 3. |
Transfer the cells to BMZERO™ supplemented with 1% FBS (1:4 mixture of base
medium to BMZERO™).
|
| 3.1. |
Once the cells are growing well with 1% serum, continue to step down the
serum concentration to 0.75% (1:6 mixture of base medium to BMZERO™)
and then to 0.5% (1:10 mixture of base medium to BMZERO™).
|
| 3.2. |
Adjust the size of the step down increments as needed to maintain good
growth. Once the cells are growing well in 0.5% serum (or 1:10 mixture of
base medium to BMZERO™ ), proceed to step 4.
|
| 4. |
Transfer the cells to BMZERO™ without serum or base medium. Passage the cells
using standard trypsinization techniques once a monolayer is observed.
|
Tips
| 1. |
For attachment-dependent cells pre-coat the
growth substratum with FNC Coating Mix®.
This becomes more important as the serum
concentration is reduced in the medium.
|
| 1. |
When performing the step down cultivation,
continue to grow a portion of the cells in the
medium from which they came to ensure an
adequate supply of adapted cells should the
cells not grow in the next step.
|
Material Safety Data
FOR RESEARCH USE ONLY. NOT INTENDED OR APPROVED
FOR HUMAN, DIAGNOSTICS OR VETERINARY
USE. Do not ingest, swallow or inhale. Do not get in
eyes, on skin, or on clothing. Wash thoroughly after
handling. For complete safety information see full
Material Safety Data Sheet.
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