Chinese Journal of Tissue Engineering Research ›› 2019, Vol. 23 ›› Issue (33): 5347-5352.doi: 10.3969/j.issn.2095-4344.1824

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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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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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