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Morphological and Biochemical Observations on Cells Derived from the in Vitro Differentiation of the Embryonal Carcinoma Cell Line PCC4-F¹

Department of Life Sciences, Faculty of Arts and Sciences [A. E. C., J. B., C. L., C. A. C.], and Department of Pathology, School of Medicine [L. E. E., H. S.], University of Pittsburgh, Pittsburgh, Pennsylvania 15261

The embryonal carcinoma cell line PCC4-F was shown to differentiate in vitro to a variety of cell types. A combination of light and electron microscopy and histochemical methods identifies or strongly suggests the presence of cardiac and skeletal muscle, fibroblasts, ciliated and nonciliated epithelial cells, endodermal yolk sac cells, neurally derived cells, and fat cells. Two cell lines were isolated from differentiated cultures. Preliminary comparisons have been made between these cell lines and the undifferentiated embryonal carcinoma cells. The derived cell lines produce an extracellular fibrous substance that gives a positive reaction to the periodate-Schiff reagent indicating a glycoprotein nature. There are significant differences in the incorporation of D-[³H]glucosamine and L-[³H]fucose into soluble extracellular proteins when the derived cell lines are compared with the embryonal carcinoma. These extracellular glycoproteins fall within narrow size classes. Enzyme studies indicate the presence of a new lactate dehydrogenase isozyme which is absent or greatly diminished in the derived cell lines. The specific activity of alkaline phosphatase is dramatically decreased in the derived cell lines, whereas those of acid phosphatase, lactate dehydrogenase, and glucose-6-phosphate dehydrogenase were not significantly different. Comparison of the derived lines with the properties of parietal endodermal yolk sac cells described by Lehman et al. (J. Cellular Physiol., 84: 13–28, 1974) suggests that these new lines are very similar if not identical in properties.

¹ This research was supported in part by Grants GM-19948 and AM-12104 from the NIH, Grant BMS74-05262 from the National Science Foundation, and Grant IN-58 from the American Cancer Society.

² To whom requests for reprints should be addressed.

Received 2/28/77. Accepted 4/ 7/77.

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