Effect of donor age on biological characteristics of adipose-derived stem cells
10.3760/cma.j.issn.1671-0290.2021.05.003
- VernacularTitle:不同年龄组脂肪干细胞生物学特性的研究
- Author:
Jiatong WANG
1
;
Yi LIU
Author Information
1. 空军军医大学第二附属医院烧伤整形科，西安 710032
- Keywords:
Stem cells;
Age groups;
Adiposetissue;
Adipose-derived stem cells;
Fat transplantation;
Wound repair
- From:Chinese Journal of Medical Aesthetics and Cosmetology
2021;27(5):361-367
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To identify the influence of age factors on biological characteristics of human adipose-derived stem cells (ADSCs), and to compare the differences in morphology, aging characteristics, differentiation potential, surface marker expression, and proliferation ability of human ADSCs with different ages and the effect of human ADSCs of different ages on proliferation and migration of fibroblasts.Methods:Adipose tissues from 27 individuals of healthy beauty treatments, aged 0-59 years, were collected from Department of Burns and Plastic Surgery, the 940th Hospital of Joint Logistics Support Force from January 2020 to December 2020. According to the age of the donor, they were divided into 6 groups. 0-9 years old group (4 cases), 10-19 years old group (4 cases), 20-29 years old group (5 cases), 30- 39 years old group (5 cases), 40-49 years old group (5 cases), and 50-59 years old group (4 cases). We obtained and cultured the ADSCs of each group, subcultured to obtain the ADSCs, and observed the morphological characteristics of each group; the proliferation ability of each group of ADSCs was detected by CCK-8 method; the differentiation potential of each group of ADSCs was detected by in vitro adipogenesis and osteogenic induction. Flow cytometry was used to detect the surface marker expression levels of each group of ADSCs, and qPCR was used to detect the expression levels of HGF, PPAR-γ, type Ⅲ collagen and DbX2 in each group. We also collected the skin tissues of a healthy person from Department of Burns and Plastic Surgery of the 940th Hospital of Joint Logistics Support Force in 2020, obtained and cultured fibroblasts, subcultured to obtain the second generation of fibroblasts, and prepared each group of ADSCs conditioned medium (ADSCs-CM); the proliferation ability of fibroblasts cultured in each group of ADSCs-CM was detected by CCK-8 method, and the migration ability of fibroblasts cultured in each group of ADSCs-CM was detected by cell scratch method. SPSS 21.0 software was used for data analysis and P<0.05 was considered statistically significant. Results:The cells of all age groups were in a typical spindle shape. The cells of the younger age group showed small nuclei and the cell morphology were uniform and similar; the cells of the older age group had larger nuclei and uneven morphology. With aging, the proliferation ability of ADSCs gradually decreased. In vitro adipogenic differentiation induction results showed that with the increase in age, the ability of ADSCs to differentiate into adipocytes gradually decreased. In vitro osteogenic differentiation induction results also demonstrated that with the increase in age, the ability of ADSCs to differentiate into osteoblasts gradually decreased. The expression rates of positive surface markers of ADSCs in each group of ADSCs were all above 93%. With aging, the expression levels of PPAR-γ, HGF, and type Ⅲ collagen decreased, and the expression levels of DbX2 showed an upward trend with age. The number of β-galactosidase staining positive cells increased with age, while the difference between groups A, B and C was not statistically significant, and the difference between groups A and D, E, and F was statistically significant ( P<0.05). The ADSCs conditioned medium of each group was able to promote the proliferation of fibroblasts, but the difference between the groups was not statistically significant. With aging, the ability of ADSCs conditioned medium to promote the migration of fibroblasts gradually decreased. Conclusions:The proliferation ability and multidirectional differentiation ability of human ADSCs and the ability to secrete a variety of cytokines that promote the proliferation and migration of fibroblasts show a downward trend, but the adipogenic differentiation ability of ADSCs declines significantly after the age of 40 years. ADSCs in all age groups could promote the proliferation and migration of fibroblasts.
