Preparation for acellular human amniotic membrane matrix and assessment the immunogenic property

doi:10.16098/j.issn.0529-1356.2016.04.021

Abstract

Abstract:

Objective To prepare of the acellular human amniotic membrane matrix (AHAM) and to assess the immunogenic property of AHAM as a scaffold for cell transplantation. Methods To prepare the AHAM, the HAM was peeled from the placenta, cut into pieces and incubated in trypsin with EDTA for decellularization. For generating cryopreserved AHAM, the fresh AHAM pieces were placed in dishes with a 1∶1 mixture of glycerol and chondroitin sulfate in MEM-NEAA and stored at -80℃. Before using, the cryopreserved AHAMs were rehydrated with sterile PBS. The expression of human leucocyte antigen in HAM and AHAM was determined by immunofluorescence. The ratio of CD4⁺and CD8⁺ T cells in mouse spleen after AHAM transplantation for four weeks was assessed by immunofluorescence and flow cytometry. Results Immunofluorescence analysis confirmed that the fresh and cryopreserved AHAM were negative for human leucocyte antigen antibody. Immunofluorescence and flow cytometry analysis confirmed that the ratio of CD4⁺and CD8⁺ T cells in mouse spleen after AHAM transplantation was not changed. Conclusion The immunogenic property of AHAM is relatively low and does not cause the T cells mediated immunological rejection after transplantation in the mouse model. These results support the potential of AHAM as a cell delivery platform to treat and manage disease. 

Key words: Acellular human amniotic membrane matrix, Immunogenicity, Transplantation, T lymphocyte, Mouse

LU Xin YUAN Jie GUO Xin-yue LI Wei-hong SHANG Hong-wei ZHANG Li-xin HUANG Xiao-wu ZHANG Hai-yan*. Preparation for acellular human amniotic membrane matrix and assessment the immunogenic property[J]. AAS, 2016, 47(4): 557-562.

References

［1］Aplin JD, Campbell S, Allen TD. The extracellular matrix of human amniotic epithelium: ultrastructure, composition and deposition ［J］. J Cell Sci, 1985, 79:119-136.
［2］Malak TM, Ockleford CD, Bell SC, et al. Confocal immunofluorescence localization of collagen types Ⅰ, Ⅲ, Ⅳ, Ⅴ and Ⅵ and their ultrastructural organization in term human fetal membranes ［J］. Placenta, 1993, 14(4):385-406.
［3］Linnala A, Balza E, Zardi L, et al. Human amnion epithelial cells assemble tenascins and three fibronectin isoforms in the extracellular matrix ［J］. FEBS Lett, 1993, 317(12):74-78.
［4］Niknejad H, Peirovi H, Jorjani M, et al. Properties of the amniotic membrane for potential use in tissue engineering ［J］. Eur Cell Mater, 2008, 15:88-99.
［5］Ricci E, Vanosi G, Lindenmair A, et al. Anti-fibrotic effects of fresh and cryopreserved human amniotic membrane in a rat liver fibrosis model ［J］. Cell Tissue Bank, 2013, 14(3):475-488. 
［6］Murphy SV, Kidyoor A, Reid T, et al. Isolation, cryopreservation and culture of human amnion epithelial cells for clinical applications ［J］. J Vis Exp, 2014, 21(94): doi:10.3791/52085.
［7］Riau AK, Beuerman RW, Lim LS, et al. Preservation, sterilization and de-epithelialization of human amniotic membrane for use in ocular surface reconstruction ［J］. Biomaterials, 2010, 31(2):216-225.
［8］Yuan J, Li W, Huang J, et al. Transplantation of human adipose stem cell-derived hepatocyte-like cells with restricted localization to liver using acellular amniotic membrane ［J］. Stem Cell Res Ther, 2015, 6:217.
［9］Park KM, Hussein KH, Hong SH, et al. Decellularized liver extracellular matrix as promising tools for transplantable bioengineered liver promotes hepatic lineage commitments of induced pluripotent stem cells ［J］. Tissue Eng Part A, 2016, 22(5-6): 449-460.
［10］Kundu J, Michaelson A, Talbot K, et al. Decellularized retinal matrix: Natural platforms for human retinal progenitor cell culture ［J］. Acta Biomater, 2016(5-6): 449-460. 
［11］Lwebuga-Mukasa JS, Thulin G, Madri JA, et al. An acellular human amnionic membrane model for in vitro culture of typeⅡ pneumocytes: the role of the basement membrane in cell morphology and function ［J］. J Cell Physiol, 1984, 121(1):215-225. 
［12］Wang ZhX, Cai WQ, Qu ChB, et al. Biocompatibility of human acellular amniotic membrane with cultured vascular smooth muscle cells in vitro ［J］. Acta Anatomica Sinica, 2008, 39(4): 598-601. (in Chinese)
王振显, 蔡文清, 瞿长宝,等. 人脱细胞羊膜与体外培养大鼠血管平滑肌细胞的生物相容性［J］. 解剖学报, 2008，39（4）: 598-601.
［13］Kubo M, Sonoda Y, Muramatsu R, et al. Immunogenicity of human amniotic membrane in experimental xenotransplantation ［J］. Invest Ophthalmol Vis Sci, 2001, 42(7):1539-1546.
［14］Lion J, Taflin C, Cross AR, et al. HLA class Ⅱ antibody activation of endothelial cells promotes Th17 and disrupts regulatory T lymphocyte expansion ［J］. Am J Transplant, 2016,16(5):1408-1420.
［15］Li C, Lin Y, Gao C, et al. Function of CD4+ T cells in CD8+ T cell mediated rejection ［J］. Zhonghua Yi Xue Za Zhi, 2014, 94(34):2690-2694.

[1]	HONG Xiao-min LI San-qiang CUI Qin-yi ZHENG Run-yue YANG Meng-li LUO Ren-li LI Qian-hui. Nuclear factor-κB signaling pathway and gender differences in alcoholic liver fibrosis [J]. Acta Anatomica Sinica, 2024, 55(1): 55-61.
[2]	YUAN Lei YANG Zhi-wei YANG Hui LIU Zheng-hai HE Jie WAN Wei. Panax notoginseng saponin relieving the inflammatory pain caused by complete Freund’s adjuvant by inhibiting the activation of astrocytes in mice [J]. Acta Anatomica Sinica, 2024, 55(1): 25-31.
[3]	MA Ting-ting CHEN Jian-hong LIU Ai-cui LI Hai-ning. Role of inhibiting lncRNA TUG1 to down-regulate nucleotide binding oligomerization domain like receptor protein 1 inflammasome in delaying the progression of Alzheimer’s disease [J]. Acta Anatomica Sinica, 2024, 55(1): 32-42.
[4]	ZOU Cheng-gong FENG Hao CHEN Bing TANG Hui SHAO Chuan SUN Mou YANG Rong HE Jia-quan . Research on the dynamic changes of neurological dysfunction and cognitive function impairment in traumatic brain injury [J]. Acta Anatomica Sinica, 2024, 55(1): 43-48.
[5]	ZHANG Qin YANG Jun-xia JIANG Qing-song. Effect of fosinopril on angiotensin converting enzyme 2 and vascular endothelial growth factor in diabetic retinopathy mice [J]. Acta Anatomica Sinica, 2023, 54(6): 628-634.
[6]	CHEN Zi-xuan SI Li-na SHU Wei-han ZHANG Xin WEI Chen-yang CHENG Lu-yang YANG Song-he QIAO Yue-bing. Mechanism of melatonin regulation of PI3K/Akt/mTOR signaling in hypothalamus delaying the initiation of puberty in female mice mice [J]. Acta Anatomica Sinica, 2023, 54(6): 644-651.
[7]	LIU Zhe-chuan LI Kun MA Shuai-nan MENG Jia-qi WANG Yan-qin. Effects of liraglutide on inflammation and mitochondrial fusion/division in Parkinson’s disease model of mice induced by paraquat [J]. Acta Anatomica Sinica, 2023, 54(6): 676-681.
[8]	LIU Ye CHU Ya-nan XU Ce-lu HE Jia-cheng SU Bing-yin TAI Hao-ran. Roscovitine rescuing neuronal loss and neuroinflammation in brain regions associated with Parkinson’s disease mice [J]. Acta Anatomica Sinica, 2023, 54(6): 635-643.
[9]	WEI Chun-chun WANG Ping LIN Fang-xing MA Xian-hua ZHANG Wei-ping XIE Zhi-fang. An improved fixation method for preparing mouse brown adipose tissue for transmission electron microscopy [J]. Acta Anatomica Sinica, 2023, 54(6): 738-743.
[10]	YAN Kun WU Xiao-lin LIU Ying-juan GE Ke-li1 REN Lei-ming LI Hong-yun . Effect of cerebrotein hydrolysate-Ⅰon intestinal microflora regulation of mice with Parkinson’s disease [J]. Acta Anatomica Sinica, 2023, 54(5): 497-504.
[11]	Ling GU ZHANG Ping SONG KeXin Na Li Hong-yu Chen Zhang Jie CONG Jing ZHAI XiaoYue GU Ling ZHANG Ping SONG Ke-xin LI Na CHEN Hong-yu ZHANG Jie CONG -Jing ZHAI Xiao-yue . Morphometry of the ureteric bud branching in the developing mouse kidney [J]. Acta Anatomica Sinica, 2023, 54(5): 593-598.
[12]	HE Ke-ke LI Yuan-di WEN Ting-hao ZHU Jie GAO Jie HU Rong HAN Feng SU Min . Erythrocyte membrane-associated protein inhibiting Th17 cell differentiation via IL-6/STAT3/ROR-γt signaling pathway in experimental autoimmune encephalomyelitis mice encephalomyelitis mice [J]. Acta Anatomica Sinica, 2023, 54(5): 538-545.
[13]	FAN Wen-juan CHEN Xu-dong CHEN Yong-fang YANG Xu-guang JIN Shao-ju ZHAO Zhi-jun DENG Jin-bo. Developmental comparison between cerebral organoids in vitro and body’s cortices in vivo [J]. Acta Anatomica Sinica, 2023, 54(4): 383-391.
[14]	ZHANG Yi-zhou LI Sha MI Shi-xiong ZUO Hong-chun ZHANG Qian-qian LI Han-lin LEI Zi-han ZHANG Dong-ze CUI Hui-xian. Effect of estradiol regulating sortilin-related receptor A expression on hippocampal spine density and synaptic protein expression via estrogen receptor of mouse [J]. Acta Anatomica Sinica, 2023, 54(3): 261-268.
[15]	LIU Ke-ting CHEN Bin-bin SHUI Yu-lian SHI Qing-ming XIAO Li. Neuroprotective effect of exogenous hydrogen sulfide and the expression of matrix metalloproteinases-9 and Caspase-1 in mice model of Parkinson’s disease [J]. Acta Anatomica Sinica, 2023, 54(3): 289-295.