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. Author manuscript; available in PMC: 2022 Feb 1.
Published in final edited form as: Cryobiology. 2020 Nov 4;98:219–232. doi: 10.1016/j.cryobiol.2020.10.017

Table 2.

Multi-step methods for CPA exposure. For each step, the solution composition is listed first followed by the exposure time in parentheses.

Max CPA Conc. Step 1 Step 2 Step 3 Step 4 Step 5 Step 6 Step 7 Step 8 Step 9
1 molal Isotonic Buffer (2.5 min) 1 molal CPA in Isotonic Buffer (Variable) Hypertonic Buffer (3 min) Isotonic Buffer (5 min) Media (20-24 h)
3 molal Isotonic Buffer (2.5 min) 1 molal CPA in Isotonic Buffer (4 min) 3 molal CPA in Isotonic Buffer (Variable) 1 molal CPA in Hypertonic Buffer (4.5 min) Hypertonic Buffer (4.5 min) Isotonic Buffer (5 min) Media (20-24 h)
5 molal Isotonic Buffer (2.5 min) 1 molal CPA in Isotonic Buffer (4 min) 3 molal CPA in Isotonic Buffer (2 min) 5 molal CPA in Isotonic Buffer (Variable) 3 molal CPA in Hypertonic Buffer (6.5 min) 1 molal CPA in Hypertonic Buffer (4.5 min) Hypertonic Buffer (4.5 min) Isotonic Buffer (5 min) Media (20-24 h)
7 molal Isotonic Buffer (2.5 min) 1 molal CPA in Isotonic Buffer (4.5 min) 3 molal CPA in Isotonic Buffer (4.5 min) 7 molal CPA in Isotonic Buffer (Variable) 3 molal CPA in Hypertonic Buffer (7 min) 1 molal CPA in Hypertonic Buffer (7 min) Hypertonic Buffer (5 min) Isotonic Buffer (5 min) Media (20-24 h)
10 molal Isotonic Buffer (2.5 min) 1 molal CPA in Isotonic Buffer (4.5 min) 3 molal CPA in Isotonic Buffer (10.5 min) 10 molal CPA in Isotonic Buffer (Variable) 3 molal CPA in Hypertonic Buffer (12.5 min) 1 molal CPA in Hypertonic Buffer (7 min) Hypertonic Buffer (5 min) Isotonic Buffer (5 min) Media (20-24 h)