Specifications
| SKU | T4034 |
|---|---|
| Species | Human (H. sapiens) |
| Tissue/Organ/Organ System | Brain |
| Cell Morphology | Multipolar |
| Applications | For Research Use Only |
| Donor Gender | Donor Info Not Disclosed |
| Donor Disease | Normal |
| Isolation Method | Selected in tyrosine free medium supplemented with a mixture of growth factors |
| Cell Type | Primary Cells |
| Growth Properties | Adherent |
| Expression Profile | tyrosine hydroxylase (TH) |
| Propagation Requirements |
Use of PriCoatTMT25 Flasks (G299) or Applied Cell Extracellular Matrix (G422) is required for cell adhesion to the culture vessels. Grow cells inECM-coated culture vessels unless otherwise specified in the Propagation Requirements below.
The base medium for this cell line is Prigrow X series medium available at abm, Cat. No. TM4034. |
| Subculture Protocol | 0 |
| Unit quantity | 5×105Â cells / 1.0 ml |
| Disclaimer | 1. All test parameters provided in the CoA are conducted using abm‘s standardized culture system and procedures. The stated values may vary under the end-user’s culture conditions. Please verify that the product is suitable for your studies by referencing published papers. All sales are final. 2. If frozen cells were received and not recovered in your lab under the exact, specified conditions (using recommended culture vessel, media, additional supplements, and atmospheric conditions), a replacement is possible at a cost (plus shipping). 3. All of abm‘s cell biology products are for research use ONLY and NOT for therapeutic/diagnostic applications. abm is not liable for any repercussions arising from the use of its cell biology product(s) in therapeutic/diagnostic application(s). Please contact a technical service representative for more information. 4. abm makes no warranties or representations as to the accuracy of the information on this site. Citations from literature and provided for informational purposes only. abm does not warrant that such information has been shown to be accurate. |
Supporting Protocol
- Important Considerations for Primary Cells
- Primary Cell Handling Instructions Upon Arrival
- Subculturing Protocol
- Freezing Protocol
- Hepatocyte Instructions
- Thawing Protocol
MSDS
QC
- T4034 Lot. RZ8055
- T4034 Lot. RZ8056
- T4034 Lot. 0083825251001
- T4034 Lot. 0083825244001
- T4034 Lot. 0083825242001
- T4034 Lot. 0083825239001
Other
What is the supply procedure?
The frozen vial of cells will be shipped in Dry ice while T25 flasks by themselves are shipped at room temperature.
Does ''Normal" mean undiseased tissue/organ?
Yes
Where is this cell line derived from?
These cells are primary cells derived from undiseased individuals. Specifically for Human Dopaminergic neuronal cells, they are >70% TH positive and differentiate into neurons when plated on poly lysine coated plates in specific growth factor containing medium. Aside from primary cells, we also offer Immortalized Human Astrocytes (SV40 or hTERT) if you wish to obtain a continuous source of cells for drug discovery, high throughput screening (HTS), drug metabolism, toxicology, or cell therapy applications. Please visit the following link http://www.abmgood.com/Immortalized-Primary-Brain-Cells.html for more immortalized brain cell lines.
When we receive the cells, should we add the full content of the vial in one T25 ECM-coated flask (cat# G299)?
Yes, and the cells will need a few days to recover after thawing. Replace the medium with fresh medium every third day to maintain optimal growth.
What are the min. and max. densities the cells should be kept at?
Split ratio should be 1:2. You can subculture whenever the cells reach 90%-100% confluency.
What is the doubling time for these cells?
Around 48 hours.
What volume of medium should be added to a T25 ECM-coated flask?
6-9ml.
What is the split ratio?
The recommended split ratio is 1:2.
What would be the concentration of trypsin used for subculturing?
0.25%
Would the cells continue dividing when they are placed in the differentiation medium?
The cells will not divide much in the differentiation medium. The cells can be kept for several days by regularly renewing the medium.
What would be the recommended amount of poly-lysine to coat T25 or T75 flasks?
The working solution of 10ug/ml Poly-lysine is made before use. Use enough solution to cover the surface of the dish/flask. Leave the cell culture ware at room temperature for 30 min – 1 hr then rinse the plates with HBSS a couple of times and the dishes can be used immediately or stored at room temperature.
How often do I need to change the media?
The media should be renewed every 2-3 days
How should I store frozen vials of primary cells? Can a frozen vial be put back into liquid nitrogen after delivery?
In most cases, a vial of primary cells shipped in dry ice (-70°C) can be placed back into liquid nitrogen and recovered at a later date by rapid thawing. However it is important to note that the viability of some sensitive cell types may be reduced by the temperature shift of such treatment, making recovery more difficult. For this reason, we recommend that cells be thawed and placed into culture as soon after receipt as possible. It is best to minimize storage time at -70°C; as this is only used for shipping the cells. ABM does not warrant the viability of cells stored at -70C after shipments have been received.
How much Pen/Strep should I add to the culture media? (Only where stated in the culturing protocol provided).
How much Pen/Strep should I add to the culture media? (Only where stated in the culturing protocol provided).
What is the recommended storage temperature?
In general, if you received: Live cells: acclimatize for 3-4 hrs at 37C, 5% CO2 and then change media afterwards. Frozen cells: Immediately place cells in liquid nitrogen or dry ice; -180C.
Following differentiation what is the percentage of TH positive cells observed?
Usually around 50-60%. Please note that the differentiation information is only to be used as a guideline only and further optimization may be required.
Recommendations for obtaining bFGF, GDNF, human EGF, Poly L Lysine, and dibutyryl cAMP?
bFGF (ABM: Cat# Z101455) GDNF (ABM: Cat# Z101055) Human EGF (ABM: Cat# Z100135) P4707-50ML (Sigma) Dibutyryl-cAMP (SCBT: sc-201567) Please note that these are only to be used as a guideline only, ABM does not warrant that these will work for your experiments and the suitability of these reagents will have to be determined by the end user.
Is it possible to freeze the cells again after thawing?
Unless specified otherwise in the datasheet, primary cell lines may be re-freezed after thawing. However, primary cells are usually not good for multiple passaging and they usually last only for up to 10-12 population doublings. Our primary cells are usually provided at passage 2, therefore they can be passaged for an additional 1-2 passages. For long term use, immortalized cells are preferred.
Once the propagation and differentiation media have been made up, what is their recommended shelf life?
The supplemented media can be stored at 4 degrees for up to 4 weeks
During differentiation, should I still change the media every 2-3 days?
This is not required unless there is a colour change observed in the media. If a colour change is observed, we recommend switching the media. If not, it is not necessary.
For differentiation, do the plates need to be coated with your Applied Cell Extracellular Matrix prior to PLL?
For differentiation of these cells, the plates of your choosing do not need to be coated with our Applied Cell Extracellular Matrix; they may be coated directly with PLL.
Can I seed frozen cells directly into collagen coated 6-well plates? Or I should seed in T25 flasks (G299) first, and then transfer to 6-well plate?
If you wish to use a 6-well plate, you can seed it directly into the plate; no need to go through T25 flask then to a 6-well plate.
Why is it important to determine the optimal seeding density?
The seeding density we recommend is for when cells are plated to a new vessel. The optimal seeding density should allow cells to attach to the surface and have room to proliferate. If you seed too little, cells may not attach well to the surface. Seeding density is important as many cells need to be in close proximity for better growth. Cell-cell interactions allow cells to communicate with each other in response to changes in their microenvironment. This ability to send and receive signals is essential for the survival of the cell. In other cases, if the seeding density is too low, cells may attach but a retardation in cell growth is observed. If you seed too high, the cells will attach but there is insufficient room for further proliferation and they will stop replicating.
How many cells are each vial?
The number of cells in a vial is lot-dependent. A Certificate of Analysis stating the cell quantity of the vial will be provided with your order.
