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

LU Xin YUAN Jie GUO Xin-yue LI Wei-hong SHANG Hong-wei ZHANG Li-xin HUANG Xiao-wu ZHANG Hai-yan*

doi:10.16098/j.issn.0529-1356.2016.04.021

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Acta Anatomica Sinica ›› 2016, Vol. 47 ›› Issue (4) : 557-562. DOI: 10.16098/j.issn.0529-1356.2016.04.021

Bioengineering

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

LU Xin¹YUAN Jie²GUO Xin-yue²LI Wei-hong¹SHANG Hong-wei¹ ZHANG Li-xin¹ HUANG Xiao-wu³ ZHANG Hai-yan ^2*

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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

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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]. Acta Anatomica Sinica. 2016, 47(4): 557-562 https://doi.org/10.16098/j.issn.0529-1356.2016.04.021

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