Establishment of a serum-free culture method for human amnion mesenchymal stem cells

Abstract

Abstract: Objective To establish a method of serum free culture for human amnion mesenthymal stem cells. Methods The hAMSCs were isolated by trypsin-collagenase two step digestion method, then cultured in serum free medium. Then immunofluorescence and WST assay were used to detect cell phenotype and cells growth. Cell migration of hAMSCs cultured in serum free medium was examined by scratch wound closure assay. The expression of growth factor was detected by real time PCR assay. Results The hAMSCs cultured in serum free medium exhibited spindle-shape characteristics, were positive for CD73, CD90, but negative for CD45 and CD34. The cell proliferation showed a U-shaped curve and the average doubling time of was 72 hours.The scratch wound closure assay demonstrated a statistically significant increase of migration as compared with those cultured in FBS.The expression of VEGF,KGF,PDGF,IGF-1 was up-regulated compared with those cultured in FBS. Conclusions A serum free culture method of hAMSCs is established, which provides an experimental evidence supporting potential application.

Key words: human amnion mesenchymal stem cells (hAMSCs), serum free culture, proliferation, migration

CLC Number:

Q331

LIU Xiao-yu, SHI Ping, PAN Bo-chen, PANG Xi-ning. Establishment of a serum-free culture method for human amnion mesenchymal stem cells[J]. Basic & Clinical Medicine, 2021, 41(8): 1181-1185.

References

[1]Fu XR,Liu G,Halim A,et al. Mesenchymal stem cell migration and tissue repair[J]. Cells,2019,8:784. doi: 10.3390/cells8080784.
[2]Liu M,Chen J,Huang H,et al. Lkb1 is an important regulator of Treg differentiation and proliferation of amniotic mesenchymal stem cells[J]. Biochem Biophys Res Commun,2020,521:434-440.
[3]Gao SY,Chen T,Hao Y,et al. Exosomal miR-135a derived from human amnion mesenchymal stem cells promotes cutaneous wound healing in rats and fibroblast migration by directly inhibiting LATS2 expression[J]. Stem Cell Res Ther, 2020,11:56. doi: 10.1186/s13287-020-1570-9.
[4]Savelli S,Trombi L, D'Alessandro D,et al. Pooled human serum: a new culture supplement for bioreactor-based cell therapies. Preliminary results[J]. Cytotherapy,2018,20:556-563.
[5]Koellensperger E,Bollinger N,Dexheimer V,et al. Choosing the right type of serum for different applications of human adipose tissue-derived stem cells: influence on proliferation and differentiation abilities[J]. Cytotherapy,2014,16:789-799.
[6]Mainzer C, Barrichello C,Debret R,et al. Insulin-transferrin-selenium as an alternative to foetal serum for epidermal equivalents[J]. Int J Cosmet Sci,2014,36:427-435.
[7]庞希宁,刘晓玉,霍双枝,等. 一种人羊膜间充质干细胞无血清培养基及其培养方法:中国. 专利号:ZL 2010 1 0252201.2[P]. 2012-10-10.
[8]Tuca AC,Ertl J,Hingerl K,et al. Comparison of matrigel and matriderm as a carrier for human amnion-derived mesenchymal stem cells in wound healing[J].Placenta,2016,48:99-103.
[9]Zisa D, Shabbir A,Suzuki G,et al. Vascular endothelial growth factor (VEGF) as a key therapeutic trophic factor in bone marrow mesenchymal stem cell-mediated cardiac repair[J]. Biochem Biophys Res Commun,2009,390:834-838.
[10]Myers TJ,Yan Y,Granero-Molto F,et al. Systemically delivered insulin-like growth factor-I enhances mesenchymal stem cell-dependent fracture healing[J]. Growth Factors, 2012,30:230-241.

[1]	XIAO Yanhong, JIANG Mingdong, LIN Yeyuan, RAN Can, LIANG Bo. Scutellarin inhibits proliferation and migration of human prostate cancer cell line PC-3 [J]. Basic & Clinical Medicine, 2024, 44(9): 1229-1235.
[2]	YAO Anjun, CHEN Lingzi, JIN Huixian. Docosahexaenoic acid inhibits proliferation of human colon cancer cell line HT-29 [J]. Basic & Clinical Medicine, 2024, 44(8): 1107-1112.
[3]	ZHENG Hong, CHEN Xiaoxia, HAN Lizhou, CHEN Miao, HUANGFU Juan. Decrease of lncRNA-RMRP expression inhibits proliferation and invasion of human lung cancer cell line A549 [J]. Basic & Clinical Medicine, 2024, 44(7): 974-978.
[4]	CAO Qingwen, QI Lin, YU Bo, TIAN Chenchen, YUAN Haining, WANG Yue. Salidroside promotes proliferation and migration of human vascular endothelial cell line EA.hy926 [J]. Basic & Clinical Medicine, 2024, 44(7): 925-930.
[5]	ZHANG Pengju, LI Jie, ZHANG Mengdi, SUN Shaoke, XU Yinzhe. KAT8 promotes the proliferation of colorectal cancer cell lines by enhancing METTL3 expression [J]. Basic & Clinical Medicine, 2024, 44(7): 931-939.
[6]	GAO Yinjie, TONG Anli. Mechanisms of cell death and proliferation of aldosterone-producing adenoma [J]. Basic & Clinical Medicine, 2024, 44(6): 758-762.
[7]	WANG Tingting, XIAO Ning, LIN Dianxin, DONG Haitao. Allicin inhibits the proliferation of oral cancer cells line CAL27 [J]. Basic & Clinical Medicine, 2024, 44(5): 673-676.
[8]	JIA Anna, GUO Jinxin, ZHANG Xuan, ZHAN Shijia, YU Yongbo, GUO Yongli, CHANG Yan. Regulation of TMEFF1 on proliferation of neuroblastoma cell lines [J]. Basic & Clinical Medicine, 2024, 44(5): 637-644.
[9]	SONG Ke, SONG Ke, LI Jingyu, MA Wei, YANG Xiaokui. Protein expression and subcellular distribution of centriolar component SAS-6 in mouse oocyte meiosis [J]. Basic & Clinical Medicine, 2024, 44(5): 651-657.
[10]	LIANG Xiangcun, WEI Xiaoyu, LIANG Jian, WANG Qing, GENG Guang. Anlotinib inhibits proliferation and promotes apoptosis of human non-small cell lung cancer cell lines through miR-16-5p/PD-1 axis [J]. Basic & Clinical Medicine, 2024, 44(4): 503-512.
[11]	LU Kai, LU Jianju, GUO Wenli, HUANG Jianqi, LI Zhihua. lncRNA VIM-AS5 expression and its effect on proliferation and migration of human breast cancer cell lines [J]. Basic & Clinical Medicine, 2024, 44(4): 447-453.
[12]	GUO Hongjiang, XU Yeting, ZHANG Diya, QIU Jiaxing, WANG Yucheng, JU Rui, GUO Lei. Effect of carboxyamidotriazole-orotate on proliferation and fatty acid anabolism of human pancreatic cancer cell lines [J]. Basic & Clinical Medicine, 2024, 44(4): 440-446.
[13]	LI Na, LI Tao, YANG Dongli, YAO Yuan. Honokiol inhibits the proliferation of human adipose-derived mesenchymal stem cells [J]. Basic & Clinical Medicine, 2024, 44(4): 483-488.
[14]	ZHANG Jing, YANG Zigeng, CAI Wenqin, CAO Weiwei, WEI Hongmei, XUE Xixi, WU Bin. Inhibition of prohibitin 2 enhances the sensitivity of non-small cell lung cancer cell line A549 to erlotinib [J]. Basic & Clinical Medicine, 2024, 44(3): 325-332.
[15]	AN Qi, QI Guangzhao, HAN Chao. Aucubin inhibits the proliferation of human hepatocellular carcinoma cells line HepG2 [J]. Basic & Clinical Medicine, 2024, 44(3): 333-338.