RFP Human Dopaminergic Neuron Cells (HDaNCs)

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Description

Name of Products RFP Human Dopaminergic Neuron Cells (HDaNCs)
Catalogue Number UBP-0057RFP
Product Format Frozen Vial
Cell Number more than 5×10[5] cells/vial
GENERAL INFORMATION: RFP 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. HDaNCs cells are provided at more than 5 x 10[5] cells/vial @ Passage 1 with more than 70% TH positive. RFP HDaNCs are grown in RFP HDaNCs medium (supplemented with 10% FBS and growth factors, UBP-44). The cells are grown in regular tissue culture dishes and Subculture 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.
CHARACTERIZATION OF THE CELLS
1. Tyrosine hydroxylase (TH) more than 70% positive by immunofluorescence
2. RFP HDaNCs are negative for HIV-1, HBV, HCV, and mycoplasma.
PRODUCT USAGE: The cells are offered for Research Use Only.
SHIPPING Frozen Vials in a Dry Ice Package.
HANDLING OF ARRIVING CELLS:  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.
PROTOCOLS FOR THAWING THE CELLS AND SUBCULTURE
A) Pre-coating of T25 flasks- Add 2ml each Universal Coating Solution  (UBP-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 UBP-01B medium, 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 Universal Detachment Solution (RT) (UBP-23) into one T25 flask; gently dispose the excessive Universal Detachment 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 800g 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).