Journal of Shandong University (Health Sciences) ›› 2019, Vol. 57 ›› Issue (11): 71-77.doi: 10.6040/j.issn.1671-7554.0.2019.345

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Isolation, culture and identification of labial stem cells from patients with dry mouth caused by maxillofacial radiotherapy

SHEN Beiyong, WANG Xiaofei, LI Junxin, ZHOU Qi   

  1. Department of Stomatology, Shenzhen Second Peoples Hospital, The First Affiliated Hospital of Shenzhen University, Shenzhen 518035, Guangdong, China
  • Published:2022-09-27

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Abstract: Objective To explore the proliferative characteristics and multi-directional differentiation potential of mesenchymal stem cells(MSCs)derived from labial glands in patients with dry mouth caused by maxillofacial radiotherapy. Methods Labial glands from the lower lip obtained during biopsy from 5 patients with dry mouth were excised under sterile conditions. MSCs were isolated and cultured using tissue adherent method. The cell proliferation curve was detected with CCK-8. Expressions of MSCs-related markers were examined in isolated cells with flow cytometry. After osteogenic differentiation, the proliferation medium group and osteogenic medium group were used as the control and osteogenic induction group, respectively. The osteogenic differentiation potential was assessed with alkaline phosphatase(ALP)staining and alizarin red staining. The mRNA expression of osteogenic-related genes was determined with real time-PCR. After adipogenic differentiation, the proliferation medium group and adipogenic medium group served as the control and adipogenic induction group, respectively. Then the adipogenic potential was observed with oil red-O staining, and the mRNA 山 东 大 学 学 报 (医 学 版)57卷11期 -沈倍勇,等.颌面部放疗致口干5例患者唇腺干细胞的分离培养与多向分化潜能鉴定 \=-expression of adipogenic-related genes were detected with real time PCR. Results MSCs were successfully obtained. CCK8 assay showed the growth curve of MSCs presented an “S” shape, which was similar to other stem cells. The surface markers of CD29, CD44, CD73, CD90 and CD105 were positively expressed, while CD34, CD45 and CD106 were negatively expressed. After 7 days of osteogenic induction, ALP staining and alizarin red staining showed strong expression of ALP and mineralized nodules in the osteogenic stem cells compared with the control group. Real time PCR results showed the expression of osteogenic-related genes in the osteogenic induction group and control group were statistically significant. After 21 days of adipogenic induction, the results of oil red-O staining showed that lipid droplets of different sizes could be seen in stem cells of osteogenic induction group. There was significant difference in the expression of adipogenic-related genes between the adipogenic induction group and control group. Conclusion MSCs can be isolated from labial glands in patients with dry mouth caused by maxillofacial radiotherapy. The MSCs have the potential for proliferation and multi-directional differentiation. The labial stem cells can be used as seed cells for salivary gland function reconstruction.

Key words: Radiotherapy, Labial glands, Stem cells, Multipotential differentiation potential

CLC Number: 

  • R783
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