Hypocalcemia is a common complication following radical surgical procedures on the thyroid and the only common way to treat it is through supplementation of vitamin D derivatives or its active metabolites and calcium preparations. Another method could be the allogeneic transplantation. The aim of the study is to develop a model for secondary prevention of permanent parathyroid insufficiency based transplanting non-immunogenic parathyroid cells or auto-transplantation of banked cells.

Ten devitalized parathyroid glands were harvested from patients during thyreoidectomy. The tissues were immediately transferred to the laboratory where enzymatic digestion was performed. The cell suspension was passed through a 100 um meshed filter and centrifuged. The cells were resuspended in cryogenic medium. ¾ volume of the suspension was cryopreserved in -190oC and ¼ volume of the suspension was plated for characterization and cultivation. Cells were maintained in DMEM 1 g/l glucose, 10% FBS and 5 ug/ml gentamicin or EpiGro. Finally, cells were fixed with 10% buffered formalin, permeabilized with 1% Tween-100 and immunostained with antibodies against Calcium sensitive receptor and intracellular parathormon (PTH-FITC).

Isolation was successful performed in all 9 of 9 cases. In all cases iImmunostaining reveals the presence of CaSR and PTH double positive parathyroid cells.

The rapidity and simplicity of the gland preparation seems to be an attractive alternative for in situ autotransplantation. Our preliminary results demonstrate the feasibility of enzymatically digested parathyroid gland cryopreservation. More human samples are needed to evaluate the reproducibility and robustness of cell isolation and cryopreservation