INTRACELLULAR MARKER FOR PURIFICATION OF STEM CELLS

摘要

An intracellular enzyme, cytosolic aldehyde dehydrogenase, expressed at high levels in primitive stem cells, appears to play an important role in allowing these cells to be resistant to cyclophosphamide. A fluorescent cell-permeable, non-polar aldehyde oxidized intracellularly by ALDH is used to identify human stem cells that are then isolated. For instance, a non-polar aldehyde, such as dansyl aminoacetaldehyde, serves as a substrate for an intracellular ALDH. Bone marrow progenitor stem cells treated with DAAA proliferate normally. Bone marrow cells expressing high levels of dansyl fluorescence after incubation with DAAA and sorted based upon dansyl fluorescence, preferably using an automated fluorescence cell sorting technique such as FACS, are substantially enriched for pluripotent hematopoietic progenitor stem cells. By this method, an enriched cellular composition comprising human lymphohematopoietic stem cells can be obtained. The cell population is capable of self-regeneration in a co-culture medium and of differentiation into various lymphoid hematopoietic lineages.