Alginate microencapsulated hepatocytes optimised for transplantation in acute liver failure.
Alginate microencapsulated hepatocytes optimised for transplantation in acute liver failure.
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DOI：
10.1371/journal.pone.0113609
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发表时间：
2014
期刊：
影响因子：
3.7
通讯作者：
Mitry RR
中科院分区：
文献类型：
--
作者：
Jitraruch S;Dhawan A;Hughes RD;Filippi C;Soong D;Philippeos C;Lehec SC;Heaton ND;Longhi MS;Mitry RR
藻酸盐微囊化人肝细胞的腹膜内移植是用于管理急性肝衰竭（ALF）的有吸引力的选择，其提供了允许天然肝再生的短期支持。本研究的主要目的是建立一个优化的方案，用于生产藻酸盐包封的人肝细胞，并评估其临床应用的适用性。使用无菌GMP级材料制备人肝细胞微珠（HMB）。我们使用不同的聚合时间和细胞密度测定了HMB的物理稳定性、细胞活力和肝细胞代谢功能。研究了HMB与外周血单个核细胞（PBMC）共培养后的免疫激活作用。用半乳糖胺诱导的ALF大鼠腹腔内移植大鼠肝细胞微珠（RMBs），使用类似的优化方案产生。测定存活率和生化特征。评价回收的微珠的形态和功能。优化的HMB具有均匀的尺寸（583.5±3.3 μm），并且使用15分钟聚合时间与10分钟和20分钟相比具有机械稳定性（p<0.001）。三维共聚焦显微镜图像显示，具有相似细胞活力的肝细胞均匀分布在HMB内。3.5×106个细胞/ml的细胞密度可提供最高的活力。在人腹水中孵育的HMB显示出比对照更好的细胞活力和功能。与单独的PBMC相比，与空微珠或肝细胞微珠共培养的PBMC没有显著的活化。与对照组相比，腹膜内移植RMBs是安全的，并且显著改善了肝损伤的严重程度（空微珠和单独的培养基; p<0.01）。回收的RMB完整且无免疫细胞粘附，并含有保留功能的活肝细胞。已经建立了生产GMP级藻酸盐包封的人肝细胞的优化方案。微珠移植在ALF中提供了有效的代谢功能。这些高质量的HMB应该适合用于临床移植。
Intraperitoneal transplantation of alginate-microencapsulated human hepatocytes is an attractive option for the management of acute liver failure (ALF) providing short-term support to allow native liver regeneration. The main aim of this study was to establish an optimised protocol for production of alginate-encapsulated human hepatocytes and evaluate their suitability for clinical use. Human hepatocyte microbeads (HMBs) were prepared using sterile GMP grade materials. We determined physical stability, cell viability, and hepatocyte metabolic function of HMBs using different polymerisation times and cell densities. The immune activation of peripheral blood mononuclear cells (PBMCs) after co-culture with HMBs was studied. Rats with ALF induced by galactosamine were transplanted intraperitoneally with rat hepatocyte microbeads (RMBs) produced using a similar optimised protocol. Survival rate and biochemical profiles were determined. Retrieved microbeads were evaluated for morphology and functionality. The optimised HMBs were of uniform size (583.5±3.3 µm) and mechanically stable using 15 min polymerisation time compared to 10 min and 20 min (p<0.001). 3D confocal microscopy images demonstrated that hepatocytes with similar cell viability were evenly distributed within HMBs. Cell density of 3.5×106 cells/ml provided the highest viability. HMBs incubated in human ascitic fluid showed better cell viability and function than controls. There was no significant activation of PBMCs co-cultured with empty or hepatocyte microbeads, compared to PBMCs alone. Intraperitoneal transplantation of RMBs was safe and significantly improved the severity of liver damage compared to control groups (empty microbeads and medium alone; p<0.01). Retrieved RMBs were intact and free of immune cell adherence and contained viable hepatocytes with preserved function. An optimised protocol to produce GMP grade alginate-encapsulated human hepatocytes has been established. Transplantation of microbeads provided effective metabolic function in ALF. These high quality HMBs should be suitable for use in clinical transplantation.
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影响因子：
3.3
作者：
Mei, Jie;Sgroi, Antonino;Buehler, Leo H.
通讯作者：
Buehler, Leo H.
影响因子：
3.3
作者：
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DOI：
10.1080/09205063.2012.735100
发表时间：
2013-06-01
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3.6
作者：
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通讯作者：
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影响因子：
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作者：
DONATO, MT;GOMEZLECHON, MJ;CASTELL, JV
通讯作者：
CASTELL, JV