Chinese Journal of Tissue Engineering Research ›› 2020, Vol. 24 ›› Issue (13): 1976-1982.doi: 10.3969/j.issn.2095-4344.2045

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Separation of adipose-derived stromal vascular fraction cells by a variety of physical methods: a comparative study

Lu Peiling1, Feng Chunchao1, Liang Miaomiao1, Wei Jiatian1, Huang Jing1, 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; 2Guangzhou Plastic Surgery Hospital of Chinese Family Physician, Guangzhou 510011, Guangdong Province, China
  • Received:2019-05-29 Revised:2019-05-30 Accepted:2019-07-15 Online:2020-05-08 Published:2020-03-07
  • Contact: Nie Yunfei, Master, Physician, Guangzhou 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:Lu Peiling, 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 Feng Chunchao, Master candidate, 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 Lu Peiling and Feng Chunchao contributed equally this work.
  • Supported by:
    the National Innovation and Entrepreneurship Training Program for College Students of China, No. 201810564021; the Natural Science Foundation of Guangdong Province, No. 2018A030313625

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Abstract:

BACKGROUND: Increasing attention has been paid to vascular components of the adipose-derived matrix and adipose-derived stem cells in tissue engineering. Existing methods for separating the vascular components of the adipose-derived matrix mainly include enzymatic and bolus injection, both of which have fatal disadvantages. 

OBJECTIVE: To search for a method for preparing adipose-derived stromal vascular fractions with high efficiency, safety, and simplicity.

METHODS: The group without any treatment was used as the negative control, and the enzymatic hydrolysis method served as the positive control. The enzymatic hydrolysis method, traditional bolus method, modified bolus method, glass beads method and built-in ultrasonic waves method were compared through cell volume, survival rate, cell fragments, cell viability, increment rate and detection of microbial infection. The enzymatic hydrolysis method and the common bolus injection method were commonly used in the separation of vascular component cells of the fat source matrix; the improved bolus method was a method obtained by improving on the basis of the ordinary bolus method; the glass bead method was to use the glass bead to oscillate. The shear force generated was obtained by adding glass beads to the fat granules and shaking at 2 500 r/min for 9 minutes to prepare stromal vascular fraction cells. Using the built-in ultrasonic method, adipose tissue was treated at 25 W for 36 seconds to obtain stromal vascular fraction cells through a cavitation effect. 

RESULTS AND CONCLUSION: (1) The size of stromal vascular fraction cells isolated by five methods showed no significant difference (P > 0.05). (2)The cell viability was lowest in the negative control group, and highest in the enzymatic hydrolysis group. The cell viability in the enzymatic hydrolysis, glass bead, and built-in ultrasonic wave groups was significantly higher than that in the modified and traditional bolus groups (P < 0.05). (3) The cell survival rate and cell proliferation rate in the enzymatic hydrolysis, glass bead, and built-in ultrasonic wave groups were significantly higher than those in the modified and traditional bolus groups (P < 0.05). (4) The cell fragmentation rate and cell apoptosis rate in the enzymatic hydrolysis, glass bead, and built-in ultrasonic wave groups were significantly lower than those in the modified and traditional bolus groups (P < 0.05). (5) These results indicate that the built-in ultrasonic method and the glass bead method are better in enriching vascular components of the adipose-derived matrix. But glass bead method adds exogenous products, so it increases the risk of pollution. Built-in ultrasonic method inserts the ultrasound probe into the adipose tissue, but as long as the ultrasound probe is thoroughly sterilized, the risk of contamination is minimized. In general, the built-in ultrasonic method and the glass bead method are superior to modified and traditional bolus methods.

Key words: physical methods, stromal vascular fraction cells, adipose tissue, ultrasonic wave, cell viability, glass bead, cell fragmentation rate, cell proliferation rate

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