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Establishment and functional identification of an immortalized human parathyroid cell line

  • JIANG Nan ,
  • LI Bo-Hou ,
  • DENG Shu-Ting ,
  • YUAN Ye ,
  • WU Qiong ,
  • WEN Si-Chun ,
  • CHEN Bai-Xi ,
  • ZHAO Ze-Wen ,
  • PENG Si-Qi ,
  • XIA Yu-Bin ,
  • TAO Yi-Ming ,
  • LIU Shuang-Xin
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  • School of Medicine, South China University of Technology, Guangzhou 510006, China; 2Department of Nephrology, Guangdong Provincial  People’s   Hospital (Guangdong Academy of Medical Sciences), Southern Medical University, Guangzhou 510080, China

Received date: 2022-10-27

  Revised date: 2023-03-25

  Online published: 2023-05-12

Abstract

Objective  To establish an immortalized human parathyroid cell line and to identify its functions.  Methods  SV40 (Simian vacuolating virus 40, SV40) large antigen was introduced into the parathyroid cells by lentivirus infection, and the transfected cells were screened by puromycin. The expressions of parathyroid hormone (PTH), calcium sensitive receptor (CaSR) and glial cells missing homolog 2, (GCM2) mRNAs and proteins in the parathyroid cells were detected by cell morphology, RT-qPCR and immunofluorescence. The reactivity of transfected cells to Ca2+ was detected by cell proliferation assay.  Results  Most of the transfected parathyroid cells were fusiform, with oval nuclei and transparent cytoplasm. RT-qPCR showed that the parathyroid cells expressed CaSR, GCM2 and PTH mRNAs. Immunofluorescence showed that the immortalized parathyroid cell line expressed parathyroid associated proteins of CaSR, GCM2 and PTH. Cell proliferation experiments showed that low calcium could promote the proliferation of parathyroid cells.  Conclusion  An immortalized parathyroid cell line was successfully established in this study, which provides an experimental basis for further exploring the pathogenesis and treatment of secondary hyperparathyroidism.

Cite this article

JIANG Nan , LI Bo-Hou , DENG Shu-Ting , YUAN Ye , WU Qiong , WEN Si-Chun , CHEN Bai-Xi , ZHAO Ze-Wen , PENG Si-Qi , XIA Yu-Bin , TAO Yi-Ming , LIU Shuang-Xin . Establishment and functional identification of an immortalized human parathyroid cell line[J]. Chinese Journal of Blood Purification, 2023 , 22(05) : 359 -363 . DOI: 10.3969/j.issn.1671-4091.2023.05.009

References


参考文献
[1] Hockham C, Bao L, Tiku A, et al. Sex differences in chronic kidney disease prevalence in Asia: a systematic review and meta-analysis[J]. Clin Kidney J, 2022,15(6):1144-1151.
[2] Younes S, Mourad N, Safwan J, et al. Chronic kidney disease awareness among the general population: tool validation and knowledge assessment in a developing country[J]. BMC Nephrol, 2022,23(1):266.
[3] Zhan Y, He X, Hong D, et al. The current status of chronic kidney disease-mineral and bone disorder management in China[J]. Sci Rep, 2022,12(1):16694.
[4] Zhao X, Niu Q, Gan L, et al. Baseline data report of the China Dialysis Outcomes and Practice Patterns Study (DOPPS). Sci Rep. 2021;11(1):873..
[5] Kobayashi S. Cardiovascular Events in Hemodialysis Patients: Challenging against Vascular Calcification[J]. Ann Vasc Dis, 2017,10(1):1-7.
[6] Moe S M. Current issues in the management of secondary hyperparathyroidism and bone disease[J]. Perit Dial Int, 2001,21 Suppl 3:S241-S246.
[7] Eddington H, Hoefield R, Sinha S, et al. Serum phosphate and mortality in patients with chronic kidney disease[J]. Clin J Am Soc Nephrol, 2010,5(12):2251-2257.
[8] Clarke B L, Brown E M, Collins M T, et al. Epidemiology and Diagnosis of Hypoparathyroidism[J]. J Clin Endocrinol Metab, 2016,101(6):2284-2299.
[9] Lin W, Li Y, Chen F, et al. Klotho preservation via histone deacetylase inhibition attenuates chronic kidney disease-associated bone injury in mice[J]. Sci Rep, 2017,7:46195.
[10] Naveh-Many T, Volovelsky O. Parathyroid Cell Proliferation in Secondary Hyperparathyroidism of Chronic Kidney Disease[J]. Int J Mol Sci, 2020,21(12).
[11] Ritter C S, Pande S, Krits I, et al. Destabilization of parathyroid hormone mRNA by extracellular Ca2+ and the calcimimetic R-568 in parathyroid cells: role of cytosolic Ca and requirement for gene transcription[J]. J Mol Endocrinol, 2008,40(1):13-21.
[12] Vulpio C, Bossola M, Di Stasio E, et al. Intra-operative parathyroid hormone monitoring through central laboratory is accurate in renal secondary hyperparathyroidism[J]. Clin Biochem, 2016,49(7-8):538-543.
[13] 李珮婷, 李刚, 柳丽丹, 等. 继发性甲状旁腺功能亢进症来源的甲状旁腺原代细胞的培养及鉴定[J]. 南方医科大学学报, 2022,42(02):238-243.
[14] 孙海晨, 刘爽, 崔叶青, 等. 成人甲状旁腺细胞的培养方法及其分泌功能研究[J]. 中国普外基础与临床杂志, 2014,21(09):1120-1123.
[15] 赵越, 王晓辉, 刘爽, 等. 成人甲状旁腺细胞的培养和鉴定[J]. 中国普外基础与临床杂志, 2017,24(01):32-36.
[16] 王小兵, 王笑云, 毛慧娟, 等. 人甲状旁腺组织体外培养及高磷的影响[J]. 中华内分泌代谢杂志, 2006(01):79-80.
[17] Maqsood M I, Matin M M, Bahrami A R, et al. Immortality of cell lines: challenges and advantages of establishment[J]. Cell Biol Int, 2013,37(10):1038-1045.
[18] Dumont J, Euwart D, Mei B, et al. Human cell lines for biopharmaceutical manufacturing: history, status, and future perspectives[J]. Crit Rev Biotechnol, 2016,36(6):1110-1122.
[19] Liu N, Guo X H, Liu J P, et al. Role of telomerase in the tumour microenvironment[J]. Clin Exp Pharmacol Physiol, 2020,47(3):357-364.
[20] 黄今, 杨融辉, 马海英. 细胞永生化机制及不同细胞永生化方法研究进展[J]. 东南大学学报(医学版), 2015,34(05):821-824.
[21] Alwin P A A, Gowri S S, Kokila V V. Immortalization of neuronal progenitors using SV40 large T antigen and differentiation towards dopaminergic neurons[J]. J Cell Mol Med, 2012,16(11):2592-2610.
[22] Li P, Wu Y, Goodwin A J, et al. Generation of a new immortalized human lung pericyte cell line: a promising tool for human lung pericyte studies[J]. Lab Invest, 2021,101(5):625-635.
[23] Promjantuek W, Chaicharoenaudomrung N, Phonchai R, et al. Transgenic Immortalization of Human Dermal Fibroblasts Mediated Through the MicroRNA/SIRT1 Pathway[J]. In Vivo, 2022,36(1):140-152.
[24] Daya-Grosjean L, Azzarone B, Maunoury R, et al. SV40 immortalization of adult human mesenchymal cells from neuroretina. Biological, functional and molecular characterization[J]. Int J Cancer, 1984,33(3):319-329.
[25] Hung C J, Yao C L, Cheng F C, et al. Establishment of immortalized mesenchymal stromal cells with red fluorescence protein expression for in vivo transplantation and tracing in the rat model with traumatic brain injury[J]. Cytotherapy, 2010,12(4):455-465.
[26] Quarles L D. Extracellular calcium-sensing receptors in the parathyroid gland, kidney, and other tissues[J]. Curr Opin Nephrol Hypertens, 2003,12(4):349-355.
[27] Quarles L D. Cinacalcet HCl: a novel treatment for secondary hyperparathyroidism in stage 5 chronic kidney disease[J]. Kidney Int Suppl, 2005(96):S24-S28.
[28] Uchiyama T, Tatsumi N, Kamejima S, et al. Hypermethylation of the CaSR and VDR genes in the parathyroid glands in chronic kidney disease rats with high-phosphate diet[J]. Hum Cell, 2016,29(4):155-161.
[29] Lau W L, Obi Y, Kalantar-Zadeh K. Parathyroidectomy in the Management of Secondary Hyperparathyroidism[J]. Clin J Am Soc Nephrol, 2018,13(6):952-961.
[30] Canaff L, Guarnieri V, Kim Y, et al. Novel Glial Cells Missing-2 (GCM2) variants in parathyroid disorders[J]. Eur J Endocrinol, 2022,186(3):351-366.
[31] Yamada T, Tatsumi N, Anraku A, et al. Gcm2 regulates the maintenance of parathyroid cells in adult mice[J]. PLoS One, 2019,14(1):e210662.
[32] 练渝, 张玲, 杨阳, 等. 继发性甲状旁腺功能亢进症大鼠Gcm2的表达研究: 中华医学会第十二次全国内分泌学学术会议[C], 中国陕西西安, 2013.
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