Terada lecture notes

Terada lecture notes - Terada et al(2002 Bone marrow cells...

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1 Terada et al. (2002) Bone marrow cells adopt the phenotype of other cells by spontaneous cell fusion. Nature 416: 542-545. A. Major Questions •What is the developmental potential of adult stem cells? •Maybe adult stem cells have more more plasticity than developmental biology dogma states •Can transdifferentiation across lineages occur? •HSC example - unexpected cell types can be derived from bone marrow in vitro and in vivo •Krauss et al: a single male-derived, donor HSC transplanted into a female mouse that then developed Y chromosome-containing, endoderm-derived, epithelial cells. •Problem with Y chromosome as a marker? •neural stem cells examples •when injected into a blastocyst, contribute to all tissues of resulting animals (like ES cell) •Furthermore, incubate the neuronal stem cell with ES in tissue culture experiment and observe differentiation of the neural stem cell into other cell types •somatic cell nuclear transfer suggests oocyte can reprogram an adult somatic nucleus B.Hypothesis •Maybe ES cells can “reprogram” other adult stem cells into a pluripotent state •“reprogramming” could be due to cell-cell interaction or production of a soluble factor •Prediction: if bone marrow cells are cultured with ES cells, the ES cells may “reprogram” the stem cells to a pluripotent type of cell that will then differentiate into many cell types
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2 C. Experimental Strategy: •Isolate bone marrow (BM) cells from GFP+, puromycin resistant female mice •Incubate female GFP+, puromycin resistant BM cells with male, puromycin sensitive ES cells (1 week) •In the presence of •LIF ( why ?) •IL3 ( why ?) •Add puromycin why ? •Maintain LIF to support growth of any new ES-like cells •Remove IL3 because hematopoeitic cells aren’t of interest anymore •Do any GFP+ cells continue to proliferate?
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Terada lecture notes - Terada et al(2002 Bone marrow cells...

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