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

Previous Articles     Next Articles

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

PDF (PC)

46

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
[1] 余梦瑶, 许晓燕, 李芳, 等. 放疗导致口腔干燥的发病机制及治疗靶点研究进展[J]. 山东医药, 2017, 57(44): 107-109. YU Mengyao, XU Xiaoyan, LI Fang, et al. Advances in research on pathogenesis and therapeutic targets of oral dryness caused by radiotherapy[J]. Shandong Medical Journal, 2017, 57(44): 107-109.
[2] 张羽, 李龄, 朱小东. 头颈部肿瘤放射性口干及唾液腺功能评价方法的研究进展[J]. 肿瘤预防与治疗, 2014, 27(4): 199-203. ZHANG Yu, LI Ling, ZHU Xiaodong. Advances in research on radioactive dry mouth and salivary gland function evaluation methods for head and neck tumors[J]. Journal of Cancer Control and Treatment, 2014, 27(4): 199-203.
[3] Kuhnt T, Jirsak N, Muller AC, et al. Quantitative and qualitative investigations of salivary gland function in dependence on irradiation dose and volume for reduction of xerostomia in patients with head-and-neck cancer[J]. Strahlenther Onkol, 2005, 181(8): 520-528.
[4] Scrimger RA, Stavrev P, Parliament MB, et al. Phenomenologic model describing flow reduction for parotid gland irradiation with intensity-modulated radiotherapy: evidence of significant recovery effect[J]. Int J Radiat Oncol Biol Phys, 2004, 60(1): 178-185.
[5] Li Y, Taylor JM, Ten HR, et al. The impact of dose on parotid salivary recovery in head and neck cancer patients treated with radiation therapy[J]. Int J Radiat Oncol Biol Phys, 2007, 67(3): 660-669.
[6] 程钢, 黄邓高, 梁颖. 骨髓间充质干细胞治疗小鼠放射性唾液腺损伤的疗效分析[J]. 临床和实验医学杂志, 2018, 17(22): 2360-2363. CHENG Gang,HUANG Denggao,LIANG Ying.The mechanism of bone marrow mesenchymal stem cells in the treatment of radiation induced salivary gland injury in mice[J]. Journal of Clinical and Experimental Medicine, 2018, 17(22): 2360-2363.
[7] 黄圣元, 颜兴. 唾液腺的发育机制及组织再生的研究进展[J]. 临床口腔医学杂志, 2018, 34(9): 573-575. HUANG Shengyuan, YAN Xing. Advances in research on developmental mechanisms and tissue regeneration of salivary glands[J]. Journal of Clinical Stomatology, 2018, 34(9): 573-575.
[8] 吕璘. 人小唾液腺间充质干细胞和上皮干/祖细胞分离鉴定及其构建生物工程化唾液腺类器官的研究[D]. 北京:北京协和医学院, 2015.
[9] 林铃, 牛道立. 下颌下腺放射性损伤及干细胞治疗的研究进展[J]. 广州医科大学学报, 2014, 42(6): 101-104. LIN Ling, NIU Daoli. Research progress on radiation injury of submandibular gland and stem cell therapy[J]. Journal of Guangzhou Medical University, 2014, 42(6): 101-104.
[10] Andreadis D, Bakopoulou A, Leyhausen G, et al. Minor salivary glands of the lips: a novel, easily accessible source of potential stem/progenitor cells[J]. Clin Oral Investig, 2014, 18(3): 847-856.
[11] Jeong J, Baek H, Kim Y, et al. Human salivary gland stem cells ameliorate hyposalivation of radiation-damaged rat salivary glands[J]. Exp Mol Med, 2013, 45(11): e58. doi: 10.1038/emm.2013.121.
[12] Liu Y, Ou M, Liu H, et al. The effect of simvastatin on chemotactic capability of SDF-1α and the promotion of bone regeneration[J]. Biomaterials, 2014, 35(15): 4489-4498.
[13] 刘云松, 吕珑薇, 周永胜, 等. 体内成骨过程中人脂肪基质细胞对新骨形成的促进作用[J]. 北京大学学报(医学版), 2012(6): 916-920. LIU Yunsong, L(¨overU)Longwei, ZHOU Yongsheng, et al. Effect of human adipose- derived stromal cells on osteogenesis in vivo[J]. Journal of Peking University(Health Sciences), 2012(6): 916-920.
[14] Rotter N, Schwarz S, Jakob M, et al. Salivary gland stem cells: Can they restore radiation-induced salivary gland dysfunction?[J]. HNO, 2010, 58(6): 556-563.
[15] Neumann Y, David R, Stiubea-Cohen R, et al. Long-term cryopreservation model of rat salivary gland stem cells for future therapy in irradiated head and neck cancer patients[J]. Tissue Eng Part C Methods, 2012, 18(9): 710-718.
[16] Maimets M, Bron R, de Haan G, et al. Similar ex vivo expansion and post-irradiation regenerative potential of juvenile and aged salivary gland stem cells[J]. Radiother Oncol, 2015, 116(3): 443-448.
[17] Schwarz S, Rotter N. Human salivary gland stem cells: isolation, propagation, and characterization[J]. Methods Mol Biol, 2012, 879: 403-442. doi: 10.1007/978-1-61779-815-3_25.
[18] 李林蔚, 李国庆, 胡劲松, 等. 小鼠唾液腺干细胞体外分离培养方法研究[J]. 现代生物医学进展, 2015, 15(32): 6230-6233. LI Linwei, LI Guoqing, HU Jinsong, et al. A method for isolation of mouse salivary gland stem cells[J]. Progress in Modern Biomedicine, 2015, 15(32): 6230-6233.
[19] Yang HW, Lin MH, Xu YZ, et al. Osteogenesis of bone marrow mesenchymal stem cells on strontium-substituted nano-hydroxyapatite coated roughened titanium surfaces[J]. Int J Clin Exp Med, 2015, 8(1): 257-264.
[20] Poldervaart MT, Wang H, van der Stok J, et al. Sustained release of BMP-2 in bioprinted alginate for osteogenicity in mice and rats[J]. PLoS One, 2013, 8(8): e72610. doi: 10.1371/journal.pone.0072610.
[1] WANG Wei, WANG Yi-Feng, ZHANG Ling-Mei, LIN Qiong-Yan, HUANG Ju. umorigenicity and invasion of side population in human ovarian epithelial cancer OVCAR3 cells [J]. JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES), 2209, 47(6): 8-11.
[2] ZHANG Hui, SHI Qing, HOU Huai-Shui, LI Dong. Culture of human cord mesenchymal stem cells using animal serum-free medium and hepatogenic differentiation [J]. JOURNAL OF SHANDONG UNIVERSITY (HEALTH SCIENCES), 2209, 47(6): 88-.
[3] ZHAO Canbin, SHAO Jiang, GUAN Donghui, QIN Ying, DING Qiang, GUO Liang, WANG Weiwei, CHEN Wei, YAN Xiaolong, ZENG Ping. Mechanism of luteolin regulating Wnt/β-catenin signal pathway in inflammatory microenvironment to promote chondrogenic differentiation of bone marrow mesenchymal stem cells [J]. Journal of Shandong University (Health Sciences), 2025, 63(7): 11-22.
[4] GUO Shuhua, FAN Yang, TIAN Feng, WANG Chuanxin, DU Lutao, LI Peilong, GUO Xing, XU Shuo. Role of microfibril-associated protein 3 in regulating mesenchymal transition of glioma stem cells [J]. Journal of Shandong University (Health Sciences), 2024, 62(6): 9-16.
[5] LIU Haixia, HUANGFU Shasha, SANG Xiaoyu, CUI Dongqing, BI Jianzhong, WANG Ping. Effects of mesenchymal stem cells on ferroptosis in experimental autoimmune encephalomyelitis mice [J]. Journal of Shandong University (Health Sciences), 2024, 62(6): 1-8.
[6] SHEN Feifei, LI Dong, LIU Linghong, JU Xiuli. Umbilical cord mesenchymal stem cells sheets treated by small intestinal submucosa promote wound healing [J]. Journal of Shandong University (Health Sciences), 2024, 62(4): 68-77.
[7] LIU Wenzhu, LIU Fuchen. Differentiation and molecular pathological analysis of motor neurons in a case of spinal muscular atrophy [J]. Journal of Shandong University (Health Sciences), 2024, 62(4): 92-100.
[8] YAN Xiaolong, QIN Ying, SHAO Jiang, CHEN Dongfeng, GUAN Donghui, ZHAO Canbin. Mechanism of curcumin regulating bone formation via the Wnt/β-catenin signaling pathway [J]. Journal of Shandong University (Health Sciences), 2024, 62(10): 76-86.
[9] WANG Cheng, ZHANG Shuhui, YAN Na, MENG Linlin, CHEN Liangzhi, LIU Qinghua. Mechanism of umbilical cord mesenchymal stem cells treating bleomycin-induced pulmonary interstitial fibrosis in mice [J]. Journal of Shandong University (Health Sciences), 2023, 61(10): 17-22.
[10] RUAN Xiangyan, CHENG Jiaojiao, DU Juan, GU Muqing. Ovarian tissue cryopreservation and preservation [J]. Journal of Shandong University (Health Sciences), 2022, 60(9): 24-30.
[11] YU Jinming, YAN Weiwei, CHEN Dawei. New practice of radio-immunotherapy for lung cancer [J]. Journal of Shandong University (Health Sciences), 2021, 59(9): 1-8.
[12] MA Yanyan, GONG Yaoqin. Application of brain organoids in investigating neurodevelopmental diseases [J]. Journal of Shandong University (Health Sciences), 2021, 59(9): 22-29.
[13] XIA Shujie, GUAN Yanjie. Progress on the activation of endogenous stem cells and functional repair in LIPUS mechanotransduction [J]. Journal of Shandong University (Health Sciences), 2021, 59(9): 37-42.
[14] WANG Zhe, LIU Yujie, MAO Qian, GUAN Peixia, BAO Qihan, LI Chengsheng, QIAO Xiaowei, PAN Qingzhong, WANG Suzhen. Evaluation of the efficacy of different regimens for early triple negative breast cancer based on the inverse probability of treatment weighting method [J]. Journal of Shandong University (Health Sciences), 2021, 59(8): 113-118.
[15] ZHANG Huayu, YIN Siyuan, LIU Jian, MA Jiaxu, SONG Ru, CAO Guoqi, WANG Yibing. Quantitative proteomic analysis of epidermal stem cells in oxygen-glucose deprivation conditions [J]. Journal of Shandong University (Health Sciences), 2021, 59(4): 17-27.
Viewed
Full text


Abstract

Cited
  Shared   
  Discussed   
No Suggested Reading articles found!
Copyright 2018 © Editorial office of Journal of Shandong University (Health Sciences)
Supported by Beijing Magtech Co.Ltd