中国组织工程研究

中国组织工程研究 ›› 2018, Vol. 22 ›› Issue (29): 4593-4600.doi: 10.3969/j.issn.2095-4344.0994

• 脂肪干细胞 adipose-derived stem cells •    下一篇

人脂肪干细胞及外泌体冻干粉的安全性

李洪超，金银鹏，王 皙，李 莉，傅青春   

  1. 上海市复旦大学附属公共卫生临床中心，上海市 201508
  • 修回日期:2018-06-12 出版日期:2018-10-18 发布日期:2018-10-18
  • 通讯作者: 傅青春，硕士，主任医师，上海市复旦大学附属公共卫生临床中心，上海市 201508
  • 作者简介:李洪超，男，1991年生，山东省菏泽市人，汉族，硕士，主要从事干细胞研究。 并列第一作者：金银鹏，男，1988年生，重庆市人，汉族，硕士，主要从事干细胞研究。
  • 基金资助:

    南京军区医学创新重大课题(14ZX01)；中国肝炎防治基金会-天晴肝病研究基金资助课题(TQGB20150104)

Safety of freeze-dried powder of human adipose-derived stem cells and its exosomes

Li Hong-chao, Jin Yin-peng, Wang Xi, Li Li, Fu Qing-chun   

  1. Shanghai Public Health Clinical Center, Fudan University, Shanghai 201508, China
  • Revised:2018-06-12 Online:2018-10-18 Published:2018-10-18
  • Contact: Fu Qing-chun, Master, Chief physician, Shanghai Public Health Clinical Center, Fudan University, Shanghai 201508, China
  • About author:Li Hong-chao, Master, Shanghai Public Health Clinical Center, Fudan University, Shanghai 201508, China. Jin Yin-peng, Master, Shanghai Public Health Clinical Center, Fudan University, Shanghai 201508, China. Li Hong-chao and Jin Yin-peng contributed equally to this work.
  • Supported by:

    the Major Medical Innovation Project of the Nanjing Military Region, No. 14ZX01; the Chinese Foundation for Hepatitis Prevention and Control-the Tianqing Foundation for Liver Diseases, No. TQGB20150104

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摘要：

文章快速阅读：

文题释义：
外泌体：是一种细胞主动分泌的大小均一、直径为50-150 nm的脂质双分子层结构囊泡，可由树突细胞、淋巴细胞、成纤维细胞、间充质干细胞和肿瘤细胞等多种不同类型细胞释放。外泌体含有多种RNA、蛋白质、脂质、DNA等物质，具有多种生物学作用，近年来引起科研界的广泛关注。
超滤法：是一种膜滤法，它以多孔薄膜作为分离介质，依靠薄膜两侧压力差作为推动力来分离溶液中不同分子质量的物质。外泌体的提取方法有多种，比如超滤法、试剂盒沉淀法、蔗糖梯度离心法、超速离心法等。超滤法具有可重复利用、经济实惠、低损伤、简单易行、设备要求低等多种优势，在外泌体的提取过程中较其他几种方法更为常用。

 

摘要
背景：随着外泌体研究的深入，越来越多的实验证实外泌体用于疾病治疗的有效性，如何稳定保存外泌体、不丧失其功能显得尤为重要。
目的：探索冻干技术与常规冻存方法保存脂肪干细胞外泌体的形态学差异，探索外泌体冻干粉用于机体注射的安全性。
方法：分离培养人脂肪干细胞并提取外泌体，将预处理的细胞及外泌体进行冻干处理，显微镜观察细胞的形态，BCA检测外泌体冻干粉的蛋白含量。血管刺激性实验、肌肉刺激性实验、溶血性实验、主动全身过敏性实验、被动皮肤过敏性实验检测外泌体冻干粉的安全性。
结果与结论：外泌体经冻干、再水化处理后仍保持膜完整性，外泌体蛋白测得量恒定；外泌体冻干粉、外泌体、脂肪干细胞冻干粉、脂肪干细胞的5种安全性实验均阴性，不存在排异反应，机体注射安全性可靠。

中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程
ORCID: 0000-0002-7141-8135(李洪超)

关键词: 脂肪干细胞, 外泌体, 冻干粉, 安全性, 干细胞

Abstract:

BACKGROUND: With the in-depth study of exosomes, increasing studies have confirmed the effectiveness of exosomes in the treatment of diseases. It is of great importance to stabilize the exosomes and not lose their functions.
OBJECTIVE: To explore the morphological differences in the exosomes of adipose-derived stem cells stored by freeze-drying technology and conventional cryopreservation and to explore the safety of exosomes for injection.
METHODS: Human adipose-derived stem cells were isolated and cultured to extract the exosomes. Then, pretreated cells and exosomes were lyophilized. Cell morphology was observed under microscope and the protein content of freeze-dried exosome powder was detected by BCA. The safety of freeze-dried exosome powder was detected through vascular stimulation test, muscle stimulation test, hemolytic test, active systemic anaphylactic test, and passive skin allergy test.
RESULTS AND CONCLUSION: The exosomes still had an intact membrane after freeze-drying and rehydration. The content of exosomal proteins was in constancy. Freeze-dried exosome powder, exosomes, freeze-dried powder of adipose-derived stem cells, and adipose-derived stem cells were all negative for five safety tests, all of which are safe and reliable for body injection and cause no rejection.

中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程

Key words: Adipose Tissue, Stem Cells, Exosomes, Freeze Drying, Tissue Engineering

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