Three-dimensional Alginate-bead Culture of Human Pituitary Adenoma Cells

Abstract

A three-dimensional culture method is described in which primary pituitary adenoma cells are grown in alginate beads. Alginate is a polymer derived from brown sea algae. Briefly, the tumor tissue is cut into small pieces and submitted to an enzymatic digestion with collagenase and trypsin. Next, a cell suspension is obtained. The tumor cell suspension is mixed with 1.2% sodium alginate and dropped into a CaCl 2 solution, and the alginate/cell suspension is gelled on contact with the CaCl 2 to form spherical beads. The cells embedded in the alginate beads are supplied with nutrients provided by the culture media enriched with 20% FBS. Three-dimensional culture in alginate beads maintains the viability of adenoma cells for long periods of time, up to four months. Moreover, the cells can be liberated from the alginate by washing the beads with sodium citrate and seeded on glass coverslips for further immunocytochemical analyses. The use of a cell culture model allows for the fixation and visualization of the actin cytoskeleton with minimal disorganization. In summary, alginate beads provide a reliable culture system for the maintenance of pituitary adenoma cells.

Pituispheres Contain Genetic Variants Characteristic to Pituitary Adenoma Tumor Tissue

“…Both, pituispheres and MSC are being used in research to investigate functionality and drug response of PA tumors (22)(23)(24)(25)33), including the development of new PA cell models (26,27). Therefore, it is highly important to evaluate whether the cellular content of the cell culture models represents original PA tumor that can be achieved by NGS using a similar approach as we have shown in our study.…”

“…Both pituispheres in stem cell-permissive medium and MSC in serum supplemented media are used for PA studies (22)(23)(24)(25), additionally other approaches like three dimensional cell cultures in alginate beads or h7H humanized PA culturing methods are developed (26,27), but how well the obtained cell cultures actually represent the tumor tissue is not clear.…”

Organoid models of the pituitary gland in health and disease

“…The aggregates could be kept in culture for several (over 3) months and were used to test effects of drugs (such as dopaminergic agonists) on the PRL secretory activity of the prolactinoma cells ( 118 ). Another approach cultured primary human pituitary tumor cells in 3D (up to 4 months) in alginate beads ( 119 ). Spheroids were derived from prolactinomas ( 65 ), GH-overproducing somatotropinomas ( 66 ), and ACTH-overproducing corticotropinomas (Cushing’s disease) ( 64 )) ( Table 1 ).…”