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| Name of Products | RFP Expressing Human Dopaminergic Neuron Cells (HDaNCs) |
| Catalogue Number | cAP-0057RFP |
| Product Format | Frozen Vial |
| Cell Number | more than 5x10[5] cells/vial |
| Human dopaminergic neuronal cells are isolated from fetal brain tissue obtained from agencies authorized to procure and distribute tissues for research. Dopaminergic neuronal cells are selected in tyrosine free medium supplemented with a mixture of growth factors. Actively growing population of cells are tested for tyrosine hydroxylase (TH) expression by immunocytochemistry. RFP-Expressing Human dopaminergic neuronal cells (RFP-HDaNCs) are selected by puromycin after HDaNCs (cAP-0057) are infected with lentiviruses expressing RFP. Cells are provided with more than 5x10[5] cells/ vial at passage 2. The cells are grown in regular tissue culture dishes and subcultured at a split ratio of 1 to 2 at confluence. We recommend not to use the cells beyond 4 passages. There may be a significant decrease in the number of tyrosine hydroxylase positive cells after 4 passages as they are not maintained in selective medium to allow the growth of only cells expressing tyrosine hydroxylase. |
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| 1. | Tyrosine hydroxylase (TH) more than 70% positive by immunofluorescence |
| 2. | HDaNCs are negative for HIV-1, HBV, HCV, and mycoplasma. |
| The cells are offered for Research Use Only. |
| Frozen Vials in a Dry Ice Package. |
| When you receive the dry ice package with cells in frozen vials, transfer the frozen vials of cells into a -80C freezer for short period storage or a liquid nitrogen tank for long- term storage. |
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| A) | Pre-coating of T25 flasks- Add 2ml each Quick Coating Solution (cAP-01) into a T25 flask to cover the whole surface of the flask, 5 mins later, dispose the excessive coating solution by aspiration and the flask is ready to be used (although solution containing other extracellular matrix, i.e. gelatin, collagen, and fibronectin, can be used, make sure to optimize the conditions in advance). |
| B) | Thaw the frozen cell vial in a 37C water bath first, and then transfer the cells into the pre-coated T25 flask with 10ml of HDaNCs Growth Medium (cAP-63), cells usually become confluent with 5-7 days. |
| C) | To passage the cells, rinse the cells in a T25 flask with 5ml HBSS (RT) twice; then add 2ml Trypsin/EDTA (RT) (cAP-23) into one T25 flask; gently dispose the excessive Trypsin/EDTA solution within 20 seconds by aspiration. |
| D) | Leave the T25 flask with the cells at RT or 37C for 1 min (most cells usually will detach from the surface within 1-2 mins; or monitor the cells under a microscope until most of cells become rounded up, and then gently tap the flask against the bench surface, and the cells will move on the surface of the flask when monitoring under microscope. |
| E) | Add 5ml Trypsin Neutralization Buffer and spin down the cells with 800RPM centrifugation for 5 mins. |
| G) | Re-suspend the cell pellet with 10 or 15ml Full medium and transfer 5 ml each into 2 or 3 pre-coated T25 flasks (for 1/2 to 1/3 subculture ratio). |
| H) | Change medium every 2 or 3days and the cells usually become confluent within 7 days (when split at a 1/3 ratio). |
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| Quick Coating Solution | cAP-01 |
| HDaNC Growth Medium | cAP-63 |
| HBSS w/o Ca2+, Mg2+ | cAP-11 |
| Cell Freezing Solution (FBS) | cAP-22 |
| Cell Freezing Solution (Non-FBS) | cAP-22B |
| Trypsin/EDTA Solution | cAP-23 |
| Trypsin Neutralization Solution | cAP-28 |
| ITS (100x) | cAP-26 |
| L-Glutamine-MAXIMUM (100x) | cAP-27 |
| Human Plasma Fibronectin Solution | cAP-42 |
| Handling human tissue derived products is potentially bio-hazardous, despite tested negative for HIV, HBV and HCV DNA; nonetheless, proper precautions must be taken to avoid inadvertent exposure. |
For customers using our primary cell products, we strongly advise you using our own medium, although similar cell growth medium can be purchased and used for our primary cell products. |
The quality of cells, in term of their growth features stated in our datasheets, will not be guaranteed if NON-ANGIO-PROTEOMIE's Cell Growth Medium is used. |
Due to the sensitive nature of primary cells and cell lines, all quality related issues about the cells products must be reported back to us within ONE month period after receiving the products, no quality warranty (i.e. replacement of cells) will be provided after the ONE-Month period. Thank you for your understanding. |
