甲状腺脱细胞生物支架的制备及鉴定

doi:10.16098/j.issn.0529-1356.2019.06.022

解剖学报 ›› 2019, Vol. 50 ›› Issue (6): 827-830.doi: 10.16098/j.issn.0529-1356.2019.06.022

甲状腺脱细胞生物支架的制备及鉴定

翁杰¹黄涛涛²王志翊^1,3,4* 陈婵⁵ 万新龙³ 周小明⁴梅劲³ 王志斌³

1.温州医科大学附属第二医院急诊科，浙江温州 325027; 2.温州医科大学第二临床医学院临床医学系，浙江温州 325000;3.温州医科大学基础医学院生物支架移植与免疫研究所，浙江温州 325000; 4.温州医科大学附属第二医院全科医学科，浙江温州 325027; 5.温州医科大学附属第一医院老年医学科，浙江温州 325000

收稿日期:2018-09-14 修回日期:2018-12-04 出版日期:2019-12-06 发布日期:2019-12-06
通讯作者: 王志翊 E-mail:wzy1063@126.com
基金资助:
国家自然科学基金资助项目;国家自然科学基金资助项目;浙江省自然科学基金资助项目;温州市科技局资助项目

Preparation and identification of thyroid gland decellularized biologic scaffold

WENG Jie¹HUANG Tao-tao² WANG Zhi-yi^1,3,4* CHEN Chan⁵ WAN Xin-long³ ZHOU Xiao-ming⁴ MEI Jin³ WANG Zhi-bin³

1.Department of Emergency Medicine, the Second Affiliated Hospital and Yuying Children’s Hospital of Wenzhou Medical University, Zhejiang Wenzhou 3250027, China; 2. The Second Clinical Medical College, Wenzhou Medical University, Zhejiang Wenzhou 325000, China; 3. Institute of Bioscaffold Transplantation and Immunology, Wenzhou Medical University, Zhejiang Wenzhou 32500, China; 4. Department of General Practice, the Second Affiliated Hospital and Yuying Children’s Hospital of Wenzhou Medical University, Zhejiang Wenzhou 325027, China; 5. Department of Geriatric Medicine, the First Affiliated Hospital of Wenzhou Medical University, Zhejiang Wenzhou 325000, China

Received:2018-09-14 Revised:2018-12-04 Online:2019-12-06 Published:2019-12-06
Contact: Zhiyi Wang E-mail:wzy1063@126.com

摘要/Abstract

摘要：

目的采用十二烷基硫酸钠（SDS）浸泡联合平板振荡制备兔甲状腺脱细胞生物支架，并对脱细胞支架进行评估。方法 20只新西兰大白兔随机分为正常对照组和脱细胞组，每组10只，脱细胞组兔甲状腺组织浸泡于1%SDS溶液后放置于37.0 ℃恒温振荡器中振荡24 h，去离子水振荡清除支架内残留去细胞试剂及DNA成分。对支架进行HE染色、DNA含量检测、扫描电子显微镜检查以及免疫荧光染色进行鉴定。结果甲状腺脱细胞支架DNA含量显著低于正常甲状腺组织（P<0.01），DNA清除率达90%以上；经检测甲状腺脱细胞支架内无DNA成分残留，同时较为完整地保存了支架3D空间结构及蛋白成分。结论浸泡法联合平板振荡法可有效清除甲状腺细胞成分，较为完整地保留支架空间结构及蛋白成分。

关键词: 脱细胞, 甲状腺, 细胞外基质, 免疫荧光, 兔

Abstract:

Objective To prepare thyroid gland decullularized biologic scaffolds by soaking with rotating and to assess the decullularized scaffolds. Methods According to random number table method, twenty New Zealand white rabbits were divided into control group and decullularized group, 10 cases in each group. The thyroid glands in decullularized group were immersed in 1% SDS and rotated at 37 ℃ for 24 hours. All thyroid gland decullularized scaffolds were rotated in deionized water to wipe off the decullularized reagent and residual DNA. HE staining, DNA concentration examination, electron microscope analysis and immunofluorescence technique were performed to indentify the characteristics of decullularized scaffolds. Results Compared to control group, the DNA concentration in the decellularized scaffold decrease significantly (P<0.01), the removal rate of DNA exceeded 90%; there were no visible cell nuclei remained, the 3D space structure and the protein components preserved well. Conclusion Soaking and rotating method can effectively wipe off the cellular components from the thyroid gland tissue, and preserve the extracellular matrix and three-dimensional structure.

Key words: Decullularization, Thyroid gland, Extracellular matrix, Immunofluorescence, Rabbit

翁杰黄涛涛王志翊陈婵万新龙周小明梅劲王志斌. 甲状腺脱细胞生物支架的制备及鉴定[J]. 解剖学报, 2019, 50(6): 827-830.

WENG Jie HUANG Tao-tao WANG Zhi-yi CHEN Chan WAN Xin-long ZHOU Xiao-ming MEI Jin WANG Zhi-bin. Preparation and identification of thyroid gland decellularized biologic scaffold[J]. Acta Anatomica Sinica, 2019, 50(6): 827-830.

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甲状腺脱细胞生物支架的制备及鉴定

Preparation and identification of thyroid gland decellularized biologic scaffold

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