The Study Of Establishing Cell Lines Of Familial Papillary Thyroid Carcinoma And Parathyroid Gland

Objective: Establishing an immortal cell line of familial papillary thyroid carcinoma(FPTC), with the purpose of exploring a new approach for studying familial non-medullary thyroid carcinoma(FNMTC). Establishing a cell line of parathyroid gland and undergoing microencapsulation of the cultured cells, with the purpose of exploring a new approach for parathyroid transplantation.Methods: Firstly, we got the specimen from a patient with FPTC, then we digested the specimen to separate the primary cells and used DMEM/F12 medium(with TSH, T3, EGF and hydrocortisone) for culture. To immortal the primary cells, the exogenous genes SV40T/TERT were transfected into the cells by two ways. RT-PCR were used to detect the expression of TPO, TG, TSHR, NIS, etc and immunofluorescence were used to detect the expression of TPO and GPC3. In order to detect the genomic mutations, we extracted the peripheral blood DNA of the patient, also the cell and tumor genome. We got the specimen from a patient with parathyroidoma, then we digested the specimen to separate the primary cells and used RPMI 1640 medium for culture. When the cells could stably grow, we undertook microencapsulation of the cultured cells. Parathyroid hormone were detected during the different cultured period.Results: The FPTC cells adhered to the plate and showed an irregular polygon shape. The cells could stably grow for six months, FPTC-S(with SV40 T transfected) passaged to p26, FPTC cells passaged to p23 and FPTC-ST(with SV40T/TERT transfected) passaged to p19. Both FPTC-S and FPTC-ST could stably express TPO, TG and TSHR in m RNA level. MLH1 R217 C mutation existed in the peripheral blood of the patient, and BRAF V600 E mutation existed in the primary cultured cells and parts of the tumor tissue. Either the primary or the immortal cells had MLH1 R217 C mutation. The primary cells separated from parathyroidoma adhered to the plate and showed two kinds of shape. The cells could stably grow and passaged to 15generation. After microencapsulating, the cultured cells could stably grow and the microencapsule stayed stable.Conclusion: This study preliminarily established an immortal cell line of familial papillary thyroid carcinoma with MLH1 R217 C and BRAF V600 E mutations. This is a good cell model for studying these mutations in FPTC. The study also preliminarily examinated the culturing method of parathyroid gland, also tried to undertake microencapsulation. This is a good trail for parathyroid transplantation.