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G2/M phase arrest and mitotic cell death by tubulin polymer-
ization and stabilization of microtubules [ 32 – 34 ]. Carboplatin
interferes with the development of the genetic material in a
cell, the DNA. This stops the cell from dividing into two new
cells and leads to cell death.- Cell culture should be undertaken in a class 2 biological safety
cabinet and disinfecting the hood cabinets and gloves with
70% ethanol should be done routinely for sterility and to pre-
vent potential contamination with human pathogens. - A tuberculin syringe is used because it is marked in frequent
small increments. - Different size and length needles are used for different pur-
poses. A 25 G × (^1) 1/2 needle (0.5 mm × 40 mm) is used for sub-
cutaneous injection of cells.
- A 25 G × 5/8 needle (0.5 mm × 16 mm) is used for intraperi-
toneal injection of cells. - A 27 G × 1/2 needle (0.4 mm × 13 mm) is used for intraperi-
toneal injection of viscous drugs. - A 30 G × 1/2 needle (0.3 mm × 13 mm) is used for intraperi-
toneal injection of aqueous drugs. - Centrifuge is used for cell pelleting.
- Hemocytometer and microscope are used for counting of via-
ble cells. - Using exponential growth phase cells for subculture and espe-
cially in final cell suspension for intraperitoneal injection is
critical for rapid cell growth in the development of peritoneal
dissemination xenograft model. - Prevent cell exposure to trypsin-EDTA longer than 10 min.
Long-term incubation with high trypsin concentration can
damage cells by striping cell surface proteins and can kill the
cells. - Presence of serum inhibits trypsin activity.
- Using the final cell suspension as soon as possible for intraperi-
toneal injection is important for keeping the cells healthy.
Using serum free medium in the final cell suspension will pre-
vent cells from sticking together to form cell aggregates and
also avoids unwanted growth effect of serum inside the mouse
peritoneal cavity. Keeping the cell suspension on ice while wait-
ing for intraperitoneal injection is also important as it will slow
down the cell’s metabolic activity and prevent apoptosis. - Use a barrier facility and sterile equipment/solutions to pre-
vent mouse infections and loss of valuable mice. Do intraperi-
toneal injections under a biosafety hood for personal protection
and mice safety.
Md Sazzad Hassan and Urs von Holzen