Culture and identification of adult human parathyroid cells_CHINESE JOURNAL OF BASES AND CLINICS IN GENERAL SURGERY

Authors：

ZHAOYue ^1,2 , WANGXiaohui ¹ , LIUShuang ¹ , SUNHaichen ¹ ,  LUOBin ^1,3

1. Department of General Surgery, Xuanwu Hospital, Capital Medical University, Beijing 100053, P.R.China;
2. Department of General Surgery, The Second People's Hospital of Shanxi Province,Taiyuan 030012, P.R.China;
3. Department of General Surgery, Beijing Tsinghua Changgung Hospital, Tsinghua University Medical Center, Beijing 102218, P.R.China;

Corresponding author：

LUOBin, Email: luobin@mail.tsinghua.edu.cn

Keywords：

parathyroid cell; cell culture; polymerase chain reaction

DOI：

10.7507/1007-9424.201605105

Video：

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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.

Citation： ZHAOYue, WANGXiaohui, LIUShuang, SUNHaichen, LUOBin. Culture and identification of adult human parathyroid cells. CHINESE JOURNAL OF BASES AND CLINICS IN GENERAL SURGERY, 2017, 24(1): 32-36. doi: 10.7507/1007-9424.201605105 Copy

1.	孙海晨, 王晓辉, 罗斌, 等. 成人甲状旁腺细胞的培养方法及其分泌功能研究. 中国普外基础与临床杂志, 2014, 21(9): 1120-1123.
2.	Moe SM. Disorders involving calcium, phosphorus, and magnesium. Prim Care, 2008, 35(2): 215-237, v-vi.
3.	孔文成, 李幼生. 维生素D在肠道中生理作用的研究进展. 医学研究生学报, 2015, 28(1): 106-109.
4.	李艳玲, 王坚. 特发性甲状旁腺功能减退并原发性醛固酮增多症的临床分析. 医学研究生学报, 2014, 27(3): 335-336.
5.	傅朝春, 徐书安, 张杰, 等. 甲状腺手术中预防甲状旁腺损伤的体会. 中国普外基础与临床杂志, 2012, 19(3): 333-334.
6.	Recombinant human parathyroid hormone (Natpara). Med Lett Drugs Ther, 2015, 57(1470): 87-88.
7.	Sollinger HW, Mack E, Cook K,et al. Allotransplantation of human parathyroid tissue without immunosuppression. Transplantation, 1983, 36(6): 599-602.
8.	Chapelle T, Meuris K, Roeyen G,et al. Simultaneous kidney-parathyroid allotransplantation from a single donor after 20 years of tetany: a case report. Transplant Proc, 2009, 41(2): 599-600.
9.	Flechner SM, Berber E, Asker M,et al. Allotransplantation of cryopreserved parathyroid tissue for severe hypocalcemia in a renal transplant recipient. Am J Transplant, 2010, 10(9): 2061-2065.
10.	高庆军, 赵代伟, 王南鹏, 等. 同种异体甲状旁腺组织移植治疗甲状旁腺功能低下的实验研究. 中国普外基础与临床杂志, 2012, 19(10): 1079-1084.
11.	Hasse C, Schlosser A, Rothmund M,et al. Parathyroid allotransplantation without immunosuppression. Lancet, 1997, 350(9087): 1296-1297.
12.	Cabané P, Gac P, Amat J,et al. Allotransplant of microen-capsulated parathyroid tissue in severe postsurgical hypoparathyroidism: a case report. Transplant Proc, 2009, 41(9): 3879-3883.
13.	Woods Ignatoski KM, Bingham EL, Frome LK,et al. Differen-tiation of precursors into parathyroid-like cells for treatment of hypoparathyroidism. Surgery, 2010, 148(6): 1186-1190.
14.	温济民, 张伟辉, 宋纯. pcDNA3.1•PTH治疗甲状旁腺功能低下症血液甲状旁腺激素变化. 中国医师杂志, 2002, 4(3): 229-233.
15.	张伟辉, 宋纯, 温济民, 等. pcDNA3.1甲状旁腺激素基因治疗甲状旁腺功能低下症的实验观察. 中华医学杂志, 2002, 82(16): 56-58.
16.	王深明, 朱易凡, 王劲松. 用重组表达体系pCKM-mPTH质粒治疗甲状旁腺功能低下症的实验研究. 中华医学杂志, 2004, 84(24): 2107-2109.
17.	温济民, 钟宇华, 黄忠, 等. pcDPG基因治疗甲状旁腺功能减退症的实验研究. 中华医学杂志, 2006, 86(4): 260-265.
18.	Lee S, Hong SW, Choi HS,et al. Experimental parathyroid hormone gene therapy using ØC31 integrase. Endocr J, 2008, 55(6): 1033-1041.
19.	Adriaansen J, Perez P, Zheng C,et al. Human parathyroid hormone is secreted primarily into the bloodstream after rat parotid gland gene transfer. Human Gene Therapy, 2011, 22(1): 84-92.
20.	Tsuji K, Kai K, Agishi T,et al. Culture of human parathyroid cells for transplantation. Transplant Proc, 1999, 31(7): 2697.
21.	Nawrot I, Sawicki A, Szmidet J,et al. Allotransplantation of cultured parathyroid progenitor cells without immunosuppression: clinical results. Transplantation, 2007, 83(6): 734-740.
22.	Moskalenko V, Ulrichs K, Kerscher A,et al. Preoperative evaluation of microencapsulated human parathyroid tissue aids selection of the optimal bioartificial graft for human parathyroid allotransplantation. Transpl Int, 2007, 20(8): 688-696.
23.	王劲松, 王深明, 陈国锐, 等. 甲状旁腺瘤组织同种移植治疗甲状旁腺功能低下症初步报告. 中华显微外科杂志, 2002, 25(2): 28-30.
24.	Lee, MK, Bae YH. Cell transplantation for endocrine disorders. Adv Drug Deliv Rev, 2000, 42(1-2): 103-120.
25.	李洪波, 王常勇, 江红. 微囊技术在生物医学中应用的研究进展. 解放军医学杂志, 2006, 31(1): 83-84.
26.	杨晓昀, 陈儿同, 张华, 等. 微囊化技术在医学领域中的应用与进展. 中国组织工程研究与临床康复, 2010, 14(29): 5483-5488.

1. 孙海晨, 王晓辉, 罗斌, 等. 成人甲状旁腺细胞的培养方法及其分泌功能研究. 中国普外基础与临床杂志, 2014, 21(9): 1120-1123.
2. Moe SM. Disorders involving calcium, phosphorus, and magnesium. Prim Care, 2008, 35(2): 215-237, v-vi.
3. 孔文成, 李幼生. 维生素D在肠道中生理作用的研究进展. 医学研究生学报, 2015, 28(1): 106-109.
4. 李艳玲, 王坚. 特发性甲状旁腺功能减退并原发性醛固酮增多症的临床分析. 医学研究生学报, 2014, 27(3): 335-336.
5. 傅朝春, 徐书安, 张杰, 等. 甲状腺手术中预防甲状旁腺损伤的体会. 中国普外基础与临床杂志, 2012, 19(3): 333-334.
6. Recombinant human parathyroid hormone (Natpara). Med Lett Drugs Ther, 2015, 57(1470): 87-88.
7. Sollinger HW, Mack E, Cook K,et al. Allotransplantation of human parathyroid tissue without immunosuppression. Transplantation, 1983, 36(6): 599-602.
8. Chapelle T, Meuris K, Roeyen G,et al. Simultaneous kidney-parathyroid allotransplantation from a single donor after 20 years of tetany: a case report. Transplant Proc, 2009, 41(2): 599-600.
9. Flechner SM, Berber E, Asker M,et al. Allotransplantation of cryopreserved parathyroid tissue for severe hypocalcemia in a renal transplant recipient. Am J Transplant, 2010, 10(9): 2061-2065.
10. 高庆军, 赵代伟, 王南鹏, 等. 同种异体甲状旁腺组织移植治疗甲状旁腺功能低下的实验研究. 中国普外基础与临床杂志, 2012, 19(10): 1079-1084.
11. Hasse C, Schlosser A, Rothmund M,et al. Parathyroid allotransplantation without immunosuppression. Lancet, 1997, 350(9087): 1296-1297.
12. Cabané P, Gac P, Amat J,et al. Allotransplant of microen-capsulated parathyroid tissue in severe postsurgical hypoparathyroidism: a case report. Transplant Proc, 2009, 41(9): 3879-3883.
13. Woods Ignatoski KM, Bingham EL, Frome LK,et al. Differen-tiation of precursors into parathyroid-like cells for treatment of hypoparathyroidism. Surgery, 2010, 148(6): 1186-1190.
14. 温济民, 张伟辉, 宋纯. pcDNA3.1•PTH治疗甲状旁腺功能低下症血液甲状旁腺激素变化. 中国医师杂志, 2002, 4(3): 229-233.
15. 张伟辉, 宋纯, 温济民, 等. pcDNA3.1甲状旁腺激素基因治疗甲状旁腺功能低下症的实验观察. 中华医学杂志, 2002, 82(16): 56-58.
16. 王深明, 朱易凡, 王劲松. 用重组表达体系pCKM-mPTH质粒治疗甲状旁腺功能低下症的实验研究. 中华医学杂志, 2004, 84(24): 2107-2109.
17. 温济民, 钟宇华, 黄忠, 等. pcDPG基因治疗甲状旁腺功能减退症的实验研究. 中华医学杂志, 2006, 86(4): 260-265.
18. Lee S, Hong SW, Choi HS,et al. Experimental parathyroid hormone gene therapy using ØC31 integrase. Endocr J, 2008, 55(6): 1033-1041.
19. Adriaansen J, Perez P, Zheng C,et al. Human parathyroid hormone is secreted primarily into the bloodstream after rat parotid gland gene transfer. Human Gene Therapy, 2011, 22(1): 84-92.
20. Tsuji K, Kai K, Agishi T,et al. Culture of human parathyroid cells for transplantation. Transplant Proc, 1999, 31(7): 2697.
21. Nawrot I, Sawicki A, Szmidet J,et al. Allotransplantation of cultured parathyroid progenitor cells without immunosuppression: clinical results. Transplantation, 2007, 83(6): 734-740.
22. Moskalenko V, Ulrichs K, Kerscher A,et al. Preoperative evaluation of microencapsulated human parathyroid tissue aids selection of the optimal bioartificial graft for human parathyroid allotransplantation. Transpl Int, 2007, 20(8): 688-696.
23. 王劲松, 王深明, 陈国锐, 等. 甲状旁腺瘤组织同种移植治疗甲状旁腺功能低下症初步报告. 中华显微外科杂志, 2002, 25(2): 28-30.
24. Lee, MK, Bae YH. Cell transplantation for endocrine disorders. Adv Drug Deliv Rev, 2000, 42(1-2): 103-120.
25. 李洪波, 王常勇, 江红. 微囊技术在生物医学中应用的研究进展. 解放军医学杂志, 2006, 31(1): 83-84.
26. 杨晓昀, 陈儿同, 张华, 等. 微囊化技术在医学领域中的应用与进展. 中国组织工程研究与临床康复, 2010, 14(29): 5483-5488.

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Culture and identification of adult human parathyroid cells

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