Method to obtain vegetable fibers from the isolation and cultivation of meristematic cells

A method for identifying and isolating exchange meristematic cells from xylem tissue, the method comprising the steps of:

• (i) taking apex segments of stems from vegetable material and submerging the apex segments in a solution of a first staining reagent;
• (ii) making tangential cuts at different heights of the stems;
• (iii) identify, under a microscope, cells dyed by the first staining reagent and identify components of xylematic tissue;
• (iv) taking the apex segments and submerging the apex segments in a solution of a second staining reagent;
• (v) making cross-sections tangential cuts at different heights of the stems;
• (vi) analyzing under the microscope cells dyed by the second staining reagent and identifying a height where meristematic cambial cells of interest are;
• (vii) making new tangential cuts of fine segments of the stems at the height where the meristematic cambial cells of interest have been identified to obtain explants;
• (viii) introducing the explants obtained in step (vii) into a liquid culture medium containing an enzyme;
• and
• (ix) verifying the presence of free cells,
• wherein the enzyme contained in the liquid culture medium of step (viii) is selected from the group consisting of pectinases, cellulases, hemicellulases, and mixtures thereof.

A method for multiplying isolated meristematic cambial cells, the method comprising the steps of:

• (i) establishing a suspension of free meristematic cambial cells in a liquid culture medium;
• (ii) maintaining a stable suspension of between 2 g/L to 15 g/L in dry weight of cells;
• and
• (iii) performing subcultures of the stable suspension of step (iii) by adding between 1 g/L and 15 g/L dry weight of cells in fresh culture medium and keeping the subcultures under permanent stirring.

A method for obtaining fiber-like structures from cultured meristematic cambial cells, the method comprising:

• (i) suspending meristematic cells in a first culture medium comprising macronutrients, micronutrients, vitamins, antioxidants, and phytohormones;
• (ii) taking and suspending the meristematic cells from step (i) again in a second culture medium comprising macronutrients, micronutrients, vitamins, antioxidants, and phytohormones;
• (iii) stirring the suspension from step (ii) for a first induction period;
• (iv) separating the cells from the second culture medium and adding a fresh amount of the second culture medium;
• and
• (v) agitating the cells and the fresh amount of the second culture medium for a second induction period,
• wherein the second culture medium comprises phytohormones different from the phytohormones of the first culture medium.