Medium-term preservation of mesophyll cells isolated from Asparagus officinalis L.: Development of a simple method by storage at reduced temperature

Abstract

Simple conditions have been investigated allowing Asparagus officinalis L. mesophyll cells to be stored as high density suspensions with unimpaired growth capacities in cell cultures. In a classic cell culture medium, storage by itself affected the mesophyll cell population by a temperature-dependent increase of mortality. Long storage reduced the ability of living cells to enter the first mitosis when used as inoculum. This loss of mitotic capacity led to reduced growth capacities (i.e. reduced increase of the number of cells) of the cultures and was relieved by decreasing the temperature of storage. Storage at 4°C in a sucrose solution and in the usual Asparagus cell culture medium has been compared. The cell culture medium containing salts, sucrose and the hormones NAA, 6-BAP gave the best results. The effective period of cell preservation is defined as the period during which cultures could be obtained with recovery of mitotic activity and growth equaling at least 75% of the control. Even in the best conditions tested, this appeared variable and dependent upon the physiological age of the cladophylls from which the cells had been obtained. It was never less than 15 days and rose to 45 days when the cells came from young cladophylls.