Biology of Reproduction, Vol 18, 141-147, Copyright © 1978 by Society for the Study of Reproduction

Separation of Deciduomal Cells by Velocity Sedimentation at Unit Gravity

¹ Departments of Obstetrics and Gynecology, Biological Chemistry and Anatomy, University of Cincinnati, College of Medicine, Cincinnati, Ohio 45267

The growth and differentiation of decidual cells proceeds as a wave of activity through the endometrial stroma during early pregnancy. Because of the diversity of decidual cells resulting from mitosis and differentiation, we have used collagenase digestion of deciduomal tissue and sedimentation of the isolated cells at unit gravity to enrich fractions of deciduomal cells which are actively synthesizing DNA, RNA, or protein. Five major fractions of cells synthesizing DNA were observed on Day 6 of pseudopregnancy and 3 of these fractions containing larger cells were active on Day 8. DNA synthesis on Day 10 was limited to low rates in larger cells. Cells synthesizing RNA were fractionated and the distributions of these cells on Days 6 and 8 of pseudopregnancy were similar. Autoradiographic examination of cell fractions was used to determine the diameter of cells actively synthesizing DNA and RNA. The distribution of deciduoma cells synthesizing protein on Day 8 was similar to the distributions of cells synthesizing RNA. Glucose-6-phosphate dehydrogenase + 6-phosphogluconate dehydrogenase activity of deciduoma cell fractions has been determined and fractions of cells with higher specific activity (enzyme units/mg protein) have been identified. These studies indicate that deciduoma cell types can be enriched by velocity sedimentation at unit gravity.

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ACKNOWLEDGMENTS This research was supported by grants from the National Institutes of Health (HD-07225 and HD-06982). We thank Beth B. Koenig and Joyce Collier for their skilled technical assistance.