Effects of different purification methods on adipocyte viability
10.3760/cma.j.issn.1009-4598.2018.01.014
- VernacularTitle: 不同纯化方式对脂肪细胞活性的影响
- Author:
Ran AN
1
;
Jiaming SUN
;
Jiecong WANG
Author Information
1. Department of Plastic Surgery, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430022, China
- Publication Type:Journal Article
- Keywords:
Adipocytes;
Cell purification;
Cell survival;
Centrifugation;
Washing with filtration;
Static precipitation
- From:
Chinese Journal of Plastic Surgery
2018;34(1):58-63
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To study the effects of different purification method on the ratio of each component in the suspension of fat granules and the cell viability.
Methods:Fat granule suspension was harvested under conventional swelling anesthesia, and divided into staticseparation group, centrifugation group, washing and filtration group.①staticseparation group: based on different gravity separation time (30 s, 1 min, 5 min, 10 min, 30 min, 1 h, 4 h, 12 h, 24 h), the group was subdivided into 9 groups.②Centrifugation group: based on different centrifugation times (3, 5, 10 min) combined withdifferent centrifugation rates (1 000, 2 000, 3 000, 4 000 r/min), this group was subdivided into 12 groups.③Washing and filtration group: According to the layers of gauze (1 layer, 2 layers, 5 layers), this group was subdivided into three groups. Adipocyte activity was measured by Calcein-AM / Hoechst 33342 dual fluorescence staining, and the viable cell rate was calculated. T test was introducedto compare between two samples. One-way AVOVA analysis was used to compare between two groups of samples.
Results:Staticseparation group: After gravity separation, the fat granules were separated intooil layer, fat granules layer and liquid layer, and stabilized at 5 min after staticseparation. ②Centrifugation group: The viable cell ratewas highest at 3 min in 2 000 r/min group.③Washing and filtration group: More volume of fat particles were collected by filtration with 2-layer gauze, and the viable cell rate was not significantly reduced.Compared with static 5 min method (86.75±6.29)%, 2 000 r/min 3 min (89.05±7.16)% and double gauze filtration rinse, the cell viability rate was (87.85±5.92)%, and there was no significant difference between the three groups (P> 0.05).
Conclusions:5 min static separation, 2-layer gauze filter and 2 000 r/min 3 min fat cell survival rate is the highest among each group, and no significant difference in viable cell rate between these three method . Clinically, 2 000 r/min 3 min is suitable for a small amount of fat filling, washing and filtration is suitable for massive fat filling.
