| abstract |
The invention discloses an amphibian cell dissociation kit. The kit of the present invention includes the following reagents: Reagent C contains collagenase I and dispase II, and the concentrations are 43000-53000U/100ml and 90-110units/100ml in sequence; Reagent D contains collagenase I, and the concentration is 55000-65000U/100ml. Reagent E contains trypsin and metal ion chelating agent, and the concentrations are 120000-180000USPU/100ml and 0.03-0.07g/100ml in sequence. Reagent F contains collagenase II at a concentration of 50000‑55000U/100ml. The invention also protects a method for dissociating amphibian tissue to obtain single cells: skin tissue, cut into pieces, and treated with reagent C; muscle tissue, cut into pieces, firstly treated with reagent D, and then treated with reagent E; bone tissue, cut into pieces crushed and treated with reagent F. The invention can be applied to the single-cell research of various amphibian limb skin, muscle and bone tissue. |