Components
PharmaOnco™ CellExpander Hematopoietic Cells Medium II (SF), 100 mL
PharmaOnco™ CD34+ Expansion Supplement (10X), 10 mL
PharmaOnco™ UM-729, 250 µg
Material not included
DMSO
Iscove's Modified Dulbecco's Medium (IMDM)
DNase I Solution
Trypan Blue
Tissue culture-treated plates
245 mm x 245 mm Square Treated Tissue Culture Dishes
35 mm Culture Dishes
Reagent Preparation
Use sterile techniques to prepare PharmaOnco™ Leukemic Cell Culture Medium. The following example is for preparing 100 mL of medium. If preparing other volumes, adjust accordingly.
1. Thaw PharmaOnco™ CellExpander Hematopoietic Cells Medium II (SF) at room temperature (15 - 25°C) or overnight at 2 - 8°C. Mix thoroughly.
2. Thaw PharmaOnco™ CD34+ Expansion Supplement (10X) at room temperature (15 - 25°C) until just thawed. Mix thoroughly.
3. Add 10 mL of PharmaOnco™ CD34+ Expansion Supplement (10X) to 90 mL of PharmaOnco™ CellExpander Hematopoietic Cells Medium II (SF). Mix thoroughly.
4. Add 250 µg of UM-729 to 68 µL of fresh DMSO (final concentration 10 mM).
5. Prepare a 1 in 100 dilution of the 10 mM UM-729 stock solution by performing serial 1 in 10 dilutions.
6. Add 10 µL of diluted UM-729 per mL of medium to give a final concentration of 1.0 µM UM-729. Titration of UM-729 may be required to determine the optimal concentration for CD34+ cell expansion.
Cell Background
Leukemic Cell
Precaution of Use
When handling biohazardous materials such as human cells, safe laboratory procedures should be followed and protective clothing should be worn.
Storage
PharmaOnco™ CellExpander Hematopoietic Cells Medium II (SF) : Store at -20°C;
PharmaOnco™ CD34+ Expansion Supplement (10X), Store at -20 to -80°C;
PharmaOnco™ UM-729, Store at 15-25°C.
Storage Comment
If not used immediately, aliquot into tubes and store at -20°C. After thawing aliquots, use immediately. Do not re-freeze.
Expiry Date
Stable as supplied until expiry date on label.
Restrictions
Restrictions for research use only. Intended for use by laboratory professionals.