Culture And Identification Of Adult Human Parathyroid Cells And Observation Of Morphology And Function Of Human Parathyroid Cells Encapsulated By Hydrogel

Objective: To investigate the reliability of culture method of adult human parathyroid cel s.Method: Adult human parathyroid tissue was digested by collagenase, then the original generation of cells were cultured and passaged, and their morphological changes were observed and recorded every other day. The cellular morphology was intact after the digestion. On the 2nd day, most of the cells had adhered and spread, and were spindle-shaped. The cells covered the flask by day 7. Different from the original generation cells, the passage cells adhered and spread on the 1st day, and were also in spindle-like shape but slightly bigger. The passaged cells covered the flask by day 7 either. Part of the passaged cells was observed through electron microscope and its supernatant PTH was assayed. Meanwhile, the other part of cells were tested the parathyroid markers, including PTH, calcium-sensing receptor(Ca SR) and GCM2 by PCR.Result: Abundant cytoplasm, mitochondria, endoplasmic reticulum and Golgi apparatus from the seventh day’s passaged cells were observed by the electron microscopy, as well as, some secretory granules existing in both cytoplasm and intercellular lacuna. Also, the PTH from supernate was detected, and parathyroid specific markers, such as Ca SR, PTH, GCM2 were positive.Conclusion: These trials demonstrated the adult human parathyroid cells could be harvested by collagenase digestion and the cultured, furthermore, the cells remained good shape and kept functioning, making it a potential source for allogeneic cell transplantation to the treatment of permanent hypoparathyroidism.Objective: To explore the feasibility of the hydrogel as a cell carrier for parathyroid cel s.Methods: Cryopreserved parathyroid cells were recovered and cultured until sufficient, then the cells were digested 、centrifuged and pipetted to single-cell suspension. The chitosan solution、crosslinking agents and cell suspension were mixed to hydrogels by some ratio. After that, hydrogels covered by complete medium were incubated at 37℃ in constant temperature incubator. Meanwhile, the supernatant were collected for measuring the PTH level every other day and the viability and shape of cells in hydrogels was also evaluated by fluorescopy.Results: Parathyroid cells were recovered and cultured successfully and the adherent cells were spindle-like. The hydrogel could be prepared at room temperature in less than 1 min. The fluorescent confocal microscopy showed that more than 90% of the cells in the hydrogel kept viable after 24 h and the proportion had no significant change after 3 days. Furthermore, PTH was detected in the supernatants.Conclusions: The facilely prepared inexpensive hydrogel, which can maintain normal morphlology and function of cells, is of potential application to cell transplantation therapy for hypopatathyroidism.