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Method of culturing pancreatic islet-like tissues by a tissue complex of pancreas-derived non-endocrinal epithelial cells and vascular endothelial cells

机译:通过胰腺来源的非内分泌上皮细胞和血管内皮细胞的组织复合物培养胰岛样组织的方法

摘要

According to embodiments, a method of producing insulin-producing tissues (IPTs) by culturing comprises: preparing non-endocrinal epithelial cells (NEECs) and vascular endothelial cells (VECs), which have been isolated or originated from postnatal pancreata, preferably by capturing of NEECs by collagen; culturing in vitro the NEECs and the VECs at least partly separately from each other; and then generating in vitro a tissue complex (IPTs) that contains both the NEECs and the VECs. In another embodiment, the native islet cells are seeded on a monolayer of VECs that have preferably been separately cultured and purified. In a further embodiment, a method of enriching NEECs comprises: culturing NEECs adhering to a container or substrate; removing NEECs by treating with a tissue-dissociation enzyme to leave left-over cells (LOCs) still attached on the container or substrate; and culturing NEECs in a medium conditioned by, or in the presence of the LOCs.
机译:根据实施方案,通过培养产生胰岛素产生组织(IPT)的方法包括:制备非内分泌上皮细胞(NEEC)和血管内皮细胞(VEC),所述非内分泌上皮细胞和血管内皮细胞(VEC)优选地通过捕获出生后胰腺而分离或起源于胰腺。胶原蛋白产生的NEEC; NEEC和VEC至少部分彼此分开体外培养;然后在体外生成包含NEEC和VEC的组织复合物(IPT)。在另一个实施方案中,将天然胰岛细胞接种在VEC的单层上,所述VEC已优选地分别培养和纯化。在另一个实施方案中,富集NEEC的方法包括:培养粘附在容器或底物上的NEEC;通过用组织解离酶处理除去NEEC,以使残留的细胞(LOC)仍附着在容器或基质上;在由LOC或存在LOC的培养基中培养NEEC。

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