利用在生物材料上培養各種人類間充質幹細胞治療移植物抗宿主疾病;GVHD treatment using several types of hMSCs cultured on biomaterials

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題名:	利用在生物材料上培養各種人類間充質幹細胞治療移植物抗宿主疾病;GVHD treatment using several types of hMSCs cultured on biomaterials
作者:	蔡長諺;Tsai, Chang-Yen
貢獻者:	化學工程與材料工程學系
關鍵詞:	移植物抗宿主病;生物材料;間質幹細胞;人類羊水幹細胞;人類脂肪幹細胞;Graft Versus Host Disease;Biomaterial;Mesenchymal Stem Cells;Human Amniotic Fluid Stem Cells;Human Adipose Derived Stem Cells
日期:	2024-07-26
上傳時間:	2024-10-09 15:25:01 (UTC+8)
出版者:	國立中央大學
摘要:	移植物抗宿主疾病（GVHD）是一種異體移植過後可能發生的重要併發症，尤其是在骨髓移植手術後最常發生。間充質幹細胞（MSCs）因其免疫抑制性能而引起人們的關注，可被用於治療GVHD等免疫相關疾病。在這項研究中，我們想評估各種類型的人類間充質幹細胞 (hMSCs) 在抑制GVHD方面的功效，並通過hMSCs研究GVHD治療的最佳細胞培養環境（生物材料）。透過hMSCs進行了成骨細胞分化以產生成骨細胞，模擬了GVHD情境中常見受影響的骨細胞環境。隨後，用異體外周血單核細胞共同培養了人羊膜液幹細胞（hAFSCs）、人脂肪來源幹細胞（hADSCs）和人類多能幹細胞（hiPSCs，HPS0077）來源的MSCs。在共同培養後，對hMSCs進行了活細胞和死細胞的檢測，以評估其對異體外周血單核細胞的活性和反應。這些細胞經過異體外周血單核細胞共同培養，評估了它們抑制與GVHD相關的炎症反應的能力。結果表明，hADSCs、hAFSCs和hiPSCs來源的MSCs能夠抑制由異體外周血單核細胞引起的炎症反應。此外，在異體外周血單核細胞存在的情況下評估了不同細胞外基質（ECM）的細胞培養環境（基質膠Matrigel、層粘連蛋白laminin-521和人類重組-玻璃粘連蛋白rVN）對hMSCs存活的影響。在塗有Matrigel的培養皿上培養的hMSCs在接受外周血單核細胞處理後比其他ECM塗層培養的細胞表現出更高的存活率。值得注意的是，在LN521上培養的hAFSCs和hiPSCs來源的MSCs表現出增強的免疫耐受性，表明這些細胞在預防免疫反應方面具有潛力。該研究突顯了這些hMSCs的免疫抑制作用的未來潛力，特別是hiPSC來源的MSCs，在治療GVHD以外的其他免疫相關疾病方面。總的來說，這些發現強調了各種hMSCs的有希望的治療潛力，以及在優化其治療GVHD的效果時考慮細胞培養環境的重要性。 ;Graft versus host disease (GVHD) is a significant complication that can occur after allogeneic transplants, particularly bone marrow transplantation. Mesenchymal stem cells (MSCs) have garnered attention for their potential in treating immune-related diseases such as GVHD because of their immunosuppressive properties. In this study, I aimed to evaluate the efficacy of various types of human MSCs (hMSCs) in suppressing GVHD, and the optimal cell culture environment (biomaterials) was investigated for GVHD treatment by hMSCs. The osteogenic differentiation of hMSCs was conducted to generate osteoblasts, which simulated the bone cell environment commonly affected in GVHD scenarios. Subsequently, human amniotic fluid stem cells (hAFSCs), human adipose-derived stem cells (hADSCs), and human pluripotent stem cells (hiPSCs, HPS0077)-derived MSCs were treated with allogeneic mononuclear cells. Following this treatment, live and dead staining of hMSCs was conducted to assess their viability and response to allogeneic mononuclear cells. These cells were subjected to allogeneic mononuclear cell treatment to evaluate their ability to suppress inflammation reactions associated with GVHD. hiPSC-derived osteoblasts with several hMSCs (hAFSCs, hADSCs, and hiPSC-derived MSCs) were also treated with allogenic mononuclear cells and were evaluated the viability of osteoblasts and the immune response by the mononuclear cells. Results indicated that hADSCs, hAFSCs, and hiPSCs-derived MSCs could suppress inflammation reactions on hMSCs as well as hiPSC-derived osteoblasts, which were induced by allogenic mononuclear cells. Furthermore, the effect of extracellular matrix (ECM) proteins (laminin-521, recombinant vitronectin, and Matrigel) on the survival of hMSCs and hiPSC-derived osteoblasts was evaluated in the presence of allogeneic mononuclear cells. hMSCs and hiPSC-derived osteoblasts cultured on Matrigel-coated dishs exhibited higher viability than those cultured on other ECM-coated dishs after treatment with allogeneic mononuclear cells. Notably, hAFSCs and hiPSCs-derived MSCs cultured on LN521 demonstrated enhanced immune tolerance, suggesting the potential for application in preventing immune reactions by the cells. The study highlights the future potential of these immunosuppressive effects of hMSCs, particularly hiPSC-derived MSCs, in treating other immune-related diseases beyond GVHD. Overall, the findings underscore the promising therapeutic potential of various hMSCs and the importance of considering the cell culture environment in optimizing their efficacy for GVHD treatment.
顯示於類別:	[化學工程與材料工程研究所] 博碩士論文

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