The cell-based therapeutics and cell fate of stem cells derived from amniotic fluid of pig and mouse conceptuses
Date Issued
2014
Date
2014
Author(s)
Peng, Shao-Yu
Abstract
Amniotic fluid-derived stem cells (AFSCs) are multipotent and shed from the fetus into the amniotic fluid. We established AFSCs from Ds-red transgenic pig and EGFP transgenic mouse which were produced in our lab for tracing and tracking purpose. The result indicated that mouse derived AFSCs can express stem cells related markers such as CD29, CD44, Sca-1, whereas do not express hematopoietic、lymphocyte、immune related or endothelial cells markers such as CD31, CD45, CD34, CD166, CD11b, CD117, CD133, CD86, CD105. Pig derived AFSCs can express stem cells related markers such as CD44, CD90, yet do not express neutrophile, macrophages, endothelial cells or lymphocyte related markers such as CD31,CD4a. AFSCs can give rise to different lineages cells such as ostoblast (ARS for calcium deposition)、chondrocyte (Toluidine blue staining for glycosaminoglycan)、adipocyte (Oil Red O staining for neutral lipid vacuoles)、spontaneously beating cardiomyocyte like cells (myosin heavy chain and cardiac troponin I antibody conjugation). After differentiation of AFSCs, the cells still remain expressing fluorescent protein. The outcome demonstrated here implies that after transplantation of the cells into the receipient, we can trace the cell fate and cell destiny. Hence, AFSCs are capable of expressing stem cell like markers and differentiating into specific cell type in the experiments in vitro. However, no study has fully investigated the potentiality and destiny of these cells in in vivo experiments. Mouse fetuses (on day 12.5-13.5 of pregnancy) were transplanted in utero with mouse or pig amniotic fluid-derived stem cells. Our results demonstrated that transplantability of AFSCs into intestine, liver and kidney ect these three germ layers were observed. By means of histological section and flow cytometry methods, allogenic transplantation for example, EGFP harboring mouse AFSCs hold the potentiality to fuse with the mouse recipient Ds-red harboring cells and eventually present in yellow color. In terms of xenotransplantation, Ds-red harboring pig AFSCs are capable of fusing with the mouse recipient EGFP harboring cells and finially present in yellow color. The pathological section results reveal that after either allotransplantion or xenotransplantaion, all organs were not found tumor formation risk. The findings may justify a clinical trial of in utero transplantation during gestation for patients who have inherited genetic disorders.
Subjects
小鼠
豬
羊水幹細胞
Type
thesis
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