Effect of DMSO exposure without cryopreservation on hematopoietic progenitor cells

Colligative cryoprotectants must be non-toxic at the high concentrations required for protection of cells from freeze-injury. Human hematopoietic stem cells are usually cryopreserved in a solution containing 1.6 molal (10% v/v) DMSO. We studied the chemical toxicity of this agent to myeloid and erythroid progenitor cells from healthy donors. No DMSO toxicity was found at concentrations of 5% or 10% at either 4 degrees or 37 degrees C for incubation durations up to 1 h. DMSO at 20% did not decrease the number of progenitor cell-derived colonies per 5 x 10(4) cells cultured, but did result in cell clumping during DMSO washout, resulting in a net loss of progenitor cells. At a concentration of 40% DMSO, a direct toxicity to hematopoietic progenitors was found. Delay in removal of DMSO after thawing of cryopreserved cells for periods up to 1 h was also non-toxic to hematopoietic progenitor cells. Direct addition of DMSO at 1% or 10% final concentration (v/v) to the culture dishes suppressed colony formation. These data suggest that DMSO is not toxic to haematopoietic progenitor cells after short-term exposure at the concentrations used for cryopreservation of marrow and peripheral blood stem cells.