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Sproull, Frederick and David, Charles N. (1979): Stem cell growth and differentiation in Hydra attenuata. I. Regulation of the self-renewal probability in multiclone aggregates. In: Journal of Cell Science, Vol. 38, No. 1: pp. 155-169 [PDF, 788kB]


Interstitial stem cells in Hydra are rapidly proliferating multipotent stem cells which continuously give rise to precursors for nerve and nematocyte differentiation. Growth of the stem cell population is controlled by the cell cycle time of the stem cells and the self-renewal probability, Ps (the fraction of stem cells in each generation which divide to yield more stem cells). In normal Hydra the stem cell generation time is 24 h and Ps = 0.6; under these conditions the stem cell population doubles in 3.5 days. In the present experiments we have systematically investigated the dependence of Ps on stem cell density. We culture stem cells in a feeder layer system consisting of aggregates of nitrogen-mustard (NM)-inactivated Hydra cells. In this system stem cell density can be varied over a wide range by changing the number of clone-forming units (CFU) added to the aggregates. We have measured the growth rate of the stem cell population and the cell cycle of stem cells in NM aggregates after 4--7 days of culture. From these data we calculate the value of Ps. The results indicate that the growth rate decreases 4-fold as the number of CFU seeded per aggregate increases from 10 to 400. Under these same conditions the cell cycle remains constant. The values of Ps calculated from these results indicate the Ps decreases from 0.75 in aggregates seeded with 10--30 CFU to 0.55 in aggregates seeded with 200--400 CFU. These results support a model in which Ps is controlled by negative feedback from neighbouring stem cells. In addition, our experiments indicate that Ps decreases during the growth of stem cell clones. When only a few stem cells are seeded in aggregates, they give rise to isolated clones distributed throughout the aggregate. Ps decreases markedly within such clones as they grow in size presumably due to increasing stem cell content of the clones. Since Ps in such isolated clones declines with growth, we infer that the local stem cell concentration is what controls Ps and that the spatial range of the negative feedback signal is short compared to the dimensions of NM aggregates.

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