Immobilization of protoplasts by anchoring to microcarriers

Abstract

Datura innoxia cell suspension-derived protoplasts were anchored to Cytodex 1 microcarriers pre-swollen in buffered concanavalin A. As many as 34 protoplasts were estimated to attach per microcarrier, in comparison to a potential 47 as determined from a model based on random anchorage. Fluorescein diacetate was used as localizing agent as well as to assess viability. When included in the swelling medium fluorescence was observed almost instantaneously, first in the protoplast at its interface with the microcarrier, and later throughout the cytoplasm. However, the dye was not conjugated with the lectin, and leakage eventually resulted in fluorescence also of non-anchored protoplasts. Fluorescein-labelled concanavalin A on the other hand permitted detection of microcarriers but not of anchored protoplasts, suggesting the use of differentially fluorescing microcarriers, as an aid in identification of fusion partners.

Milestones in Plant Tissue Culture Systems for the Production of Secondary Products

“…Attention has recently been given to immobilizing plant cells to prolong their production phase and their total product yield (73). The product would have to be leaked from the plant cells, unless their immobilized protoplasts (74), organelles (75,76), or cell-free enzymes (77) were active.…”