A New Culture Technique: An Approach to the in-Vitro Reconstitution of Endocrine Organs
A new cell culture technique called three-dimensional-rotation culture (3dR-culture) has been developed. Its main characteristics are: (i) cultivation under constant rotation, and (ii) the use of a three-dimensional material that allows the cells to adhere and to grow in a three-dimensional arrangement. Two glassfibre materials and one gelatine-sponge proved suitable for the use in cell culture. The new technique has the advantage to yield in large numbers cells bound on a stable matrix that can easily be handled, e.g. for implantation.
The technique described was used for primary culture of dissociated pituitaries m combination with a um-free medium. Under these conditions a pituitary “organoid” reconstituted in-vitro which secreted in average 50 ng ACTH/ml × day into the culture supernatant. The pituitary organoids were implanted subcutaneously into hypophysectomized syngeneic rats. The influence an restoring growth and ACTH blood plasma level was observed. The animals did not increase their body weight, but the ACTH level rose from zero to at least one fifth of normal values.
Hypothalamic organoids, also grown in serum-free medium, secreted vasopressin for up to seven weeks into the culture supernatant.
These experiments show that it may be possible to substitute dysfunctional organs by in-vitro reconstituted organoids, which would open new possibilities for medical therapy and for studying the regulation of endocrine glands.
KeywordsPituitary Cell Hypothalamic Cell Glass Fibre Material Gelatine Sponge Aggregate Cell Culture
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