The United States of America, as Represented by the Secretary, Department of Health and Human Services

Other

Ron McKay

Lorenz Studer

Barbara J Wold

James V Sullivan

John Comstock Doyle

Marie E Csete

Rea Ravin

David W Munno

Daniel Hoeppner

An apparatus and process for monitoring migratory cell proliferation with restricted migration on a substrate includes providing a substrate, coating the substrate with extracellular matrix, plating cells suspended in cell culture media on extracellular matrix, and placing intersecting channels across the extracellular matrix components by removing the extracellular matrix components from the channels to isolate islands of the extracellular matrix components on the substrate. When the cells are immersed with a fluid, migration of the cells is confined to the isolated islands of the extracellular matrix components, permitting long-term observation of a migratory population.

Method of etching islands of cells in a grid pattern

The invention provides a method of culturing cells which includes a proliferating step in which the number of precursor cells is expanded and a differentiating step in which the expanded precursor cells develop into neuronal cells. The proliferating step includes the step of incubating the precursor cells in proliferating medium which includes basic fibroblast growth factor (bFGF). The differentiating step includes incubating the precursor cells in differentiation media in a manner effective to form a cellular aggregate that is not adhered to any surface of the incubation vessel. In a preferred embodiment, the cells arc incubated in a roller tube. The differentiation media can also include at least one differentiating agent. The invention also provides a method for treating a neurological disorder, such as Parkinson's disease, a method of introducing a gene product into a brain of a patient, an assay for neurologically active substances, and a cell culture.

Cell expansion system for use in neural transplantation

The present invention relates to the growth of cells in culture under conditions that promote cell survival, proliferation, and/or cellular differentiation. The present inventors have found that proliferation was promoted and apoptosis reduced when cells were grown in lowered oxygen as compared to environmental oxygen conditions traditionally employed in cell culture techniques. Further, the inventors found that differentiation of precursor cells to specific fates also was enhanced in lowered oxygen where a much greater number and fraction of dopaminergic neurons were obtained when mesencephalic precursors were expanded and differentiated in lowered oxygen conditions. Thus at more physiological oxygen levels the proliferation and differentiation of CNS precursors is enhanced, and lowered oxygen is a useful adjunct for ex vivo generation of specific neuron types. Methods and compositions exploiting these findings are described.

Low oxygen culturing of central nervous system progenitor cells

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Bethesda, MD, United States of America

Ron McKay