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.	?孫海晨, 王曉輝, 羅斌, 等. 成人甲狀旁腺細胞的培養方法及其分泌功能研究. 中國普外基礎與臨床雜志, 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. 維生素D在腸道中生理作用的研究進展. 醫學研究生學報, 2015, 28(1): 106-109.
4.	李艷玲, 王堅. 特發性甲狀旁腺功能減退并原發性醛固酮增多癥的臨床分析. 醫學研究生學報, 2014, 27(3): 335-336.
5.	傅朝春, 徐書安, 張杰, 等. 甲狀腺手術中預防甲狀旁腺損傷的體會. 中國普外基礎與臨床雜志, 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.	高慶軍, 趙代偉, 王南鵬, 等. 同種異體甲狀旁腺組織移植治療甲狀旁腺功能低下的實驗研究. 中國普外基礎與臨床雜志, 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.	溫濟民, 張偉輝, 宋純. pcDNA3.1?PTH治療甲狀旁腺功能低下癥血液甲狀旁腺激素變化. 中國醫師雜志, 2002, 4(3): 229-233.
15.	張偉輝, 宋純, 溫濟民, 等. pcDNA3.1甲狀旁腺激素基因治療甲狀旁腺功能低下癥的實驗觀察. 中華醫學雜志, 2002, 82(16): 56-58.
16.	王深明, 朱易凡, 王勁松. 用重組表達體系pCKM-mPTH質粒治療甲狀旁腺功能低下癥的實驗研究. 中華醫學雜志, 2004, 84(24): 2107-2109.
17.	溫濟民, 鐘宇華, 黃忠, 等. pcDPG基因治療甲狀旁腺功能減退癥的實驗研究. 中華醫學雜志, 2006, 86(4): 260-265.
18.	Lee S, Hong SW, Choi HS,et al. Experimental parathyroid hormone gene therapy using ?C31 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.	王勁松, 王深明, 陳國銳, 等. 甲狀旁腺瘤組織同種移植治療甲狀旁腺功能低下癥初步報告. 中華顯微外科雜志, 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, 江紅. 微囊技術在生物醫學中應用的研究進展. 解放軍醫學雜志, 2006, 31(1): 83-84.
26.	楊曉昀, 陳兒同, 張華, 等. 微囊化技術在醫學領域中的應用與進展. 中國組織工程研究與臨床康復, 2010, 14(29): 5483-5488.

1. ?孫海晨, 王曉輝, 羅斌, 等. 成人甲狀旁腺細胞的培養方法及其分泌功能研究. 中國普外基礎與臨床雜志, 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. 維生素D在腸道中生理作用的研究進展. 醫學研究生學報, 2015, 28(1): 106-109.
4. 李艷玲, 王堅. 特發性甲狀旁腺功能減退并原發性醛固酮增多癥的臨床分析. 醫學研究生學報, 2014, 27(3): 335-336.
5. 傅朝春, 徐書安, 張杰, 等. 甲狀腺手術中預防甲狀旁腺損傷的體會. 中國普外基礎與臨床雜志, 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. 高慶軍, 趙代偉, 王南鵬, 等. 同種異體甲狀旁腺組織移植治療甲狀旁腺功能低下的實驗研究. 中國普外基礎與臨床雜志, 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. 溫濟民, 張偉輝, 宋純. pcDNA3.1?PTH治療甲狀旁腺功能低下癥血液甲狀旁腺激素變化. 中國醫師雜志, 2002, 4(3): 229-233.
15. 張偉輝, 宋純, 溫濟民, 等. pcDNA3.1甲狀旁腺激素基因治療甲狀旁腺功能低下癥的實驗觀察. 中華醫學雜志, 2002, 82(16): 56-58.
16. 王深明, 朱易凡, 王勁松. 用重組表達體系pCKM-mPTH質粒治療甲狀旁腺功能低下癥的實驗研究. 中華醫學雜志, 2004, 84(24): 2107-2109.
17. 溫濟民, 鐘宇華, 黃忠, 等. pcDPG基因治療甲狀旁腺功能減退癥的實驗研究. 中華醫學雜志, 2006, 86(4): 260-265.
18. Lee S, Hong SW, Choi HS,et al. Experimental parathyroid hormone gene therapy using ?C31 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. 王勁松, 王深明, 陳國銳, 等. 甲狀旁腺瘤組織同種移植治療甲狀旁腺功能低下癥初步報告. 中華顯微外科雜志, 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, 江紅. 微囊技術在生物醫學中應用的研究進展. 解放軍醫學雜志, 2006, 31(1): 83-84.
26. 楊曉昀, 陳兒同, 張華, 等. 微囊化技術在醫學領域中的應用與進展. 中國組織工程研究與臨床康復, 2010, 14(29): 5483-5488.

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Culture and identification of adult human parathyroid cells

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