Position specific growth regulation of 3T3 cells in vivo

In this study we have investigated the mechanism by which spatial growth is regulated by monitoring 3T3 cells, introduced into the developing mouse limb using exo utero surgery. The 3T3 cells were labeled with a human cell surface glycoprotein, CD8, and injected into stage 7-9 mouse limbs. At 24 and 48 hr after injection embryos were labeled with [3H]thymidine and processed for immunohistochemistry and autoradiography. The labeling index of CD8 positive cells was compared to that of neighboring limb bud cells and also to the position of the injection site within the limb. We find that the labeling index of 3T3 cells is in accord with that of the limb cells that immediately surround them; 3T3 cells display a high labeling index in limb regions of high growth and a low labeling index in limb regions of low growth. In addition, we find that both limb bud cells and injected 3T3 cells display a general proximal (low) to distal (high) gradient of growth at the stages analyzed. We conclude from these results that position-specific regulation of growth occurs in a non-cell autonomous manner and is likely to be mediated by mitogenic signals that are localized within the limb environment. In addition, our results demonstrate the usefulness of utilizing established cell lines as in vivo probes to monitor developmental mechanisms.