中国组织工程研究

中国组织工程研究 ›› 2019, Vol. 23 ›› Issue (33): 5347-5352.doi: 10.3969/j.issn.2095-4344.1824

• 干细胞基础实验 basic experiments of stem cells • 上一篇    下一篇

不同非接触式超声波处理获得脂肪源基质血管组分细胞方法的比较

魏嘉田1，卢沛伶1，黄  婧1，柳  雪1，蔡海明1，巫国辉2，张玲华1，聂云飞2   

  1. 1广东省农业生物蛋白质功能与调控重点实验室，华南农业大学生命科学学院，广东省广州市  510642；2广州中家医家庭医生整形美容医院，广东省广州市  510011
  • 修回日期:2019-06-10 出版日期:2019-11-28 发布日期:2019-11-28
  • 通讯作者: 聂云飞，医学硕士，医师，广州中家医家庭医生整形美容医院，广东省广州市 510011； 张玲华，博士，教授，广东省农业生物蛋白质功能与调控重点实验室，华南农业大学生命科学学院，广东省广州市 510642
  • 作者简介:魏嘉田，男，1999年生，广西壮族自治区贺州市人，汉族，就读于华南农业大学生命科学学院，主要从事免疫抗衰老方向研究。
  • 基金资助:

    国家级大学生创新创业训练计划(201810564021)，项目负责人：卢沛伶；广东省自然科学基金(2018A030313625)，项目负责人：张玲华

Comparison of different non-contact ultrasound methods for obtaining adipose-derived stromal vascular fraction cells

Wei Jiatian1, Lu Peiling1, Huang Jing1, Liu Xue1, Cai Haiming1, Wu Guohui2, Zhang Linghua1, Nie Yunfei2   

  1. 1Guangdong Provincial Key Laboratory of Protein Function and Regulation in Agricultural Organisms, College of Life Sciences, South China Agricultural University, Guangzhou 510642, Guangdong Province, China; 2Plastic Surgery Hospital of Chinese Family Physician, Guangzhou 510011, Guangdong Province, China
  • Revised:2019-06-10 Online:2019-11-28 Published:2019-11-28
  • Contact: Nie Yunfei, Master, Physician, Plastic Surgery Hospital of Chinese Family Physician, Guangzhou 510011, Guangdong Province, China. Zhang Linghua, MD, Professor, Guangdong Provincial Key Laboratory of Protein Function and Regulation in Agricultural Organisms, College of Life Sciences, South China Agricultural University, Guangzhou 510642, Guangdong Province, China
  • About author:Wei Jiatian, Guangdong Provincial Key Laboratory of Protein Function and Regulation in Agricultural Organisms, College of Life Sciences, South China Agricultural University, Guangzhou 510642, Guangdong Province, China
  • Supported by:

    the National Students Innovation and Entrepreneurship Training Program, No. 201810564021 (to LPL); the Natural Science Foundation of Guangdong Province, No. 2018A030313625 (to ZLH)

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摘要：

文章快速阅读：

文题释义：
脂肪源基质血管组分：是指源于脂肪组织的血管(或涉及携带血液的脉管)成分的细胞组分，是脂肪组织去除成熟脂肪细胞后剩余的多种细胞及基质成分的组合，主要包含脂肪来源干细胞、造血干细胞、内皮祖细胞、巨噬细胞等。
非接触式超声波：非接触式超声波法具有安全、便捷、无额外添加物等特质，可用于批量处理脂肪移植材料，同时非接触式超声波破碎无需接触材料，可以有效减少污染风险，而且可以实现自动化，无需人力进行操控，显著减少人为误差，故非接触式超声波破碎法制备生物材料更加快速、有效，在干细胞疾病治疗领域具有广阔的应用前景。

 

摘要
背景：探索能够高效、稳定获取基质血管组分细胞的方法对于临床应用有非常重要的价值。目前分离基质血管组分的方法主要包括酶解法、推注破碎法、接触式超声波破碎法等，存在安全性和细胞存活率低、材料污染等不足，而非接触式超声波破碎法则能很好的解决这个问题。
目的：寻找一种最优的非接触式超声波处理获得脂肪源基质血管组分细胞的方法。
方法：实验所用人体脂肪组织(皮下脂肪)由广州中家医家庭医生整形美容医院提供，体检无其他基础疾病，取材前均获知情同意并签署知情同意书。以3种不同的超声波仪器(超声波清洗仪功率约为800 W、超声波细胞破碎仪功率约为1 000 W、非接触式超声波细胞破碎仪功率约为1 200 W)对脂肪材料进行非接触式超声处理15，20，25 min，以接触式超声波细胞破碎法为对照，功率为100 W，超声处理16 s，分别检测基质血管组分细胞大小、存活数量、细胞活力、细胞碎片率和微生物感染度。该研究实施符合广州中家医家庭医生整形美容医院的相关伦理要求。
结果与结论：①3种方法所获得的基质血管组分细胞活性均高于接触式超声波破碎法(P < 0.05)；②3种方法所获得的基质血管组分细胞污染度均低于接触式超声波破碎法；③3种方法获得的基质血管组分细胞大小差异无显著性意义(P > 0.05)；④3种方法所获得的基质血管组分细胞碎片均少于接触式超声波破碎法(P < 0.05)，其中非接触式超声波细胞破碎仪处理20 min后采用酶解方法获取基质血管组分细胞的效果最好。


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: 0000-0003-1692-6038(张玲华)

关键词: 脂肪组织, 基质血管组分, 非接触式超声波, 纯物理破碎, 细胞活力

Abstract:

BACKGROUND: It is of great clinical significance to explore methods for efficient and stable access to stromal vascular component cells. Current methods for separating stromal vascular components mainly include enzymatic hydrolysis, bolus injection method, and contact ultrasonic pulverization method. For contact ultrasonic pulverization, there are many problems to be solved, such as low safety, low cell survival rate, and material contamination. Therefore, non-contact ultrasonic pulverization can solve these problems well.
OBJECTIVE: To find an optimal method for harvesting adipose-derived stromal vascular fraction cells by non-contact ultrasound approach.
METHODS: The human body fat tissue (subcutaneous fat) used was provided by the Plastic Surgery Hospital of Chinese Family Physician. Patients were eligible if they had no other basic diseases in the physical examination. All the patients were informed of study protocol and signed informed consent before sampling. The fat tissues were treated with three different ultrasonic instruments (approximately 800 W for ultrasonic cleaner, 1 000 W for ultrasonic cell crusher, and 1 200 W for non-contact ultrasonic cell crusher) for 15, 20, and 25 minutes. Contact ultrasonic crushing method (100 W, 16 seconds) was used as control. Stromal vascular fraction cell size, number of survived cells, cell viability, cell fragmentation rate and microbial infection were measured. The study was implemented in accordance with the relevant ethical requirements of Plastic Surgery Hospital of Chinese Family Physician.
RESULTS AND CONCLUSION: The viability of stromal vascular fraction cells obtained by the three methods was significantly higher than that by the contact ultrasonic crushing method (P < 0.05). The contamination degree of stromal vascular fraction cells obtained by the three methods was significantly lower than that obtained by the contact ultrasonic crushing method (P < 0.05). There was no significant difference in the size of stromal vascular fraction cells obtained by the three methods (P > 0.05). The cell fragments of stromal vascular fraction cells obtained by the three methods were significantly less than those by contact ultrasonic crushing method (P < 0.05). It is preferred to perform 20 minutes treatment by non-contact ultrasonic cell crusher followed by enzymatic separation.

Key words: adipose tissue, stromal vascular component, non-contact ultrasonic, pure physical disruption, cell viability

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