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]	LIU Lei, LI Shi-bao, ZHANG Hao-liang. Lnc-SLC2A12-10:1 silencing inhibits the proliferation, migration and invasion of gastric cancer cell line AGS [J]. Basic & Clinical Medicine, 2022, 42(7): 1071-1076.
[2]	ZHONG Ming, YU Xiu-yi, GUO Wei-xi, FANG Zheng. Ophiopogonin B inhibits proliferation and migration of human esophageal squamous cell carcinoma cell line EC9706 [J]. Basic & Clinical Medicine, 2022, 42(7): 1053-1059.
[3]	KANG You-li, ZHAO Dan, DU Wen-jing, LI Li. SIRT5 promotes the proliferation of human breast cancer cells by regulating the succinylation of ALDH5A1 [J]. Basic & Clinical Medicine, 2022, 42(7): 1099-1107.
[4]	BAI Bing, ZHANG Hai-fang, YANG Qing-liang, QIN Yue, ZHOU Chen-long, SONG Ge, WANG Ying. Down-regulation of lncRNA RHPN1-AS1 inhibits proliferation and migration of human bladder cancer cell line T24 [J]. Basic & Clinical Medicine, 2022, 42(6): 940-944.
[5]	DU Xue-mei, ZHANG Yun-mei, ZHOU Wen-ting, LYU Hong. miR-655-3p targeting KIF20A inhibits migration and invasion of lung adenocarcinoma cell line A549 [J]. Basic & Clinical Medicine, 2022, 42(4): 599-606.
[6]	SUN Quan-peng, FAN Chao, ZHAN De-xi, FAN Yi-ming, SUN Wei-feng. Ropivacaine inhibits proliferation, migration and invasion of human liver cancer cell line Hep3B [J]. Basic & Clinical Medicine, 2022, 42(3): 448-453.
[7]	ZHANG Jian, ZHANG Ji-jiong. LncRNA UNC5B-AS1 targeting miR-339-5p regulates the proliferation and apoptosis of human lung adenocarcinoma cell line A549 [J]. Basic & Clinical Medicine, 2022, 42(3): 454-460.
[8]	LIU Xin-bing, ZHANG Mei-hong. Gelsemine inhibits proliferation and invasion of rheumatoid arthritis synovial fibroblast through down-regulating circ_001680 [J]. Basic & Clinical Medicine, 2022, 42(3): 429-435.
[9]	ZHAO Xia, CAI Qing-chun, DING Rui-min, ZHANG Qian. Cinobufagin inhibits proliferation of human nasopharyngeal carcinoma cell lines CNE2 and HONE1 [J]. Basic & Clinical Medicine, 2022, 42(3): 436-440.
[10]	CHEN Qian, LIU Xian, CUI Nan, ZHAI Yang. Knockout of EZH2 inhibits migration and invasion of human cervical cancer cell lines [J]. Basic & Clinical Medicine, 2022, 42(3): 395-400.
[11]	ZHANG Chun-li, QI Jie-hua, HUAN Yu, WU Shou-le, CAI Xu-shan. Interference with ADAM17 inhibits proliferation of human cervical cancer cell line HeLa [J]. Basic & Clinical Medicine, 2022, 42(2): 255-259.
[12]	WANG Bu, YUAN Sheng-fang, ZHANG Chang-hong, YUAN Cheng, HUANG Pan-deng, ZHANG Zhi-hua, ZHAO Jian-qing. Silencing eEF2K combined with sodium tanshinone ⅡA sulfonate synergistically inhibit proliferation of human lung adenocarcinoma cell line A549 [J]. Basic & Clinical Medicine, 2022, 42(1): 106-113.
[13]	LI Yong-xiao, SUN Yan, ZHANG Rui-sheng. Propofol inhibits proliferation of human endometrial cancer cell line RL95-2 [J]. Basic & Clinical Medicine, 2022, 42(1): 114-119.
[14]	WANG Ling-xia, YAN Yu-lan, YUAN Hai-tao, CAI Ye-wei, LIU De-hui, GU Wen-lu, CAO Bi-yue. CircRNA_23113 inhibits the proliferation and migration of lung adenocarcinoma cells [J]. Basic & Clinical Medicine, 2022, 42(1): 82-88.
[15]	SHI Tie-wei, LI Tian-zhu, ZHOU Jing, NA Ri-su, SUN Qi, XU Li-yan, BAI Chun-ying. Effects of 5-lipoxygenase activating protein inhibitor MK886 on proliferation and apoptosis of human esophageal cancer cell lines [J]. Basic & Clinical Medicine, 2022, 42(1): 56-61.