Objective To investigate the reliability of culture method of adult human parathyroid cells. Methods 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. Part of the passaged cells were observed through electron microscope and its supernatant parathyroid hormone (PTH) was assayed. Meanwhile, the other part of cells were tested the parathyroid markers, including PTH, calcium-sensing receptor (CaSR) and glial cells missing-2 (GCM-2) by PCR. Results 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 CaSR, PTH, and GCM2 were positive. Conclusions 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 culture primary parathyroid cells by mean of simulated microgravity, observe their basic morphological characteristics, study survival rate and secretory function of parathyroid cells, and explore more excellent culture mean of parathyroid cells. Methods There were 37 male Wistar rats, the body weight was 150–200 g. The rat was intraperitoneally injected with 1% pentobarbital sodium (50 mg/kg). The parathyroid glands were surgically excised and identified pathologically. The parathyroid gland cells were got and digested them with collagenase Ⅱ, which were divided into three groups: conventional culture group (simple parathyroid cells were cultured), polyglycolic acid (PGA) scaffold culture group (the parathyroid cells were cultured on the PGA scaffold), and simulated microgravity culture group (the parathyroid cells and PGA scaffolds were cultured in simulated microgravity environment). The parathyroid cells were cultured for 1, 3, 5 or 7 days in different culture conditions, then the parathyroid hormone (PTH) was measured, morphological characteristics of the parathyroid cell was observed under phase contrast microscope, survival rate of the parathyroid cells was calculated by acridine orange/propidium iodide staining. Results The parathyroid cell morphologies of most cells were well and center of part of cell mass was necrosis on day 7 in the conventional culture group. The most parathyroid cells were spreading toward the poles along the PGA cell scaffold in the longitudinal direction and the adjacent stents were connected by extracellular matrix on day 7 in the PGA scaffold culture group. The parathyroid cells cultured under the simulated microgravity were got round and formed clusters on day 7 in the simulated microgravity culture group. Compared with the other two groups on day 7, the PTH and the survival rate of the parathyroid cells were significantly higher in the simulated microgravity culture group (P<0.05). Conclusions Parathyroid cells cultured in simulated microgravity environment could maintain better morphology, survival rate is higher, and secretory function is better. Therefore, parathyroid cells cultured in simulated microgravity could be used as good donor cell for treatment of hypoparathyroidism. PGA scaffold could be used as a good carrier for culture of parathyroid cell.