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1.发明授权Method of culturing pancreatic islet-like tissues by a tissue complex of pancreas-derived non-endocrinal epithelial cells and vascular endothelial cells 有权通过胰腺衍生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公开(公告)号：US08852934B2
公开(公告)日：2014-10-07
申请号：US13635630
申请日：2011-03-15
CPC分类号： C12N5/0677 , C12N5/0676 , C12N2502/28
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.
摘要翻译： 根据实施方案，通过培养产生产生胰岛素的组织（IPT）的方法包括：制备已经从出生后胰腺分离或产生的非内分泌上皮细胞（NEEC）和血管内皮细胞（VEC），优选通过捕获 胶原蛋白NEECs; NEECs和VECs体外至少部分分离培养; 然后在体外产生包含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优选单独培养和纯化的VEC单层上。 在进一步的实施方案中，富集NEEC的方法包括：培养附着于容器或基质的NEEC; 通过用组织解离酶处理以留下仍然附着在容器或底物上的左侧细胞（LOC）来去除NEEC; 并在由LOC或在LOC的存在条件下培养NEECs。
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2.发明申请METHOD OF CULTURING PANCREATIC ISLET-LIKE TISSUES BY A TISSUE COMPLEX OF PANCREAS-DERIVED NON-ENDOCRINAL EPITHELIAL CELLS AND VASCULAR ENDOTHELIAL CELLS 有权通过胰腺衍生的非内皮细胞上皮细胞和血管内皮细胞的组织复合物培养胰蛋白酶样组织的方法公开(公告)号：US20130122586A1
公开(公告)日：2013-05-16
申请号：US13635630
申请日：2011-03-15
IPC分类号： C12N5/071
CPC分类号： C12N5/0677 , C12N5/0676 , C12N2502/28
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.
摘要翻译： 根据实施方案，通过培养产生产生胰岛素的组织（IPT）的方法包括：制备已经从出生后胰腺分离或产生的非内分泌上皮细胞（NEEC）和血管内皮细胞（VEC），优选通过捕获 胶原蛋白NEECs; NEECs和VECs体外至少部分分离培养; 然后在体外产生包含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优选单独培养和纯化的VEC单层上。 在进一步的实施方案中，富集NEEC的方法包括：培养附着于容器或基质的NEEC; 通过用组织解离酶处理以留下仍然附着在容器或底物上的左侧细胞（LOC）来去除NEEC; 并在由LOC或在LOC的存在条件下培养NEECs。
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3.发明申请Cryopreservation medium and cryopreservation method for tissues and cells 审中-公开冷冻保存培养基和组织和细胞的低温保存方法公开(公告)号：US20070259327A1
公开(公告)日：2007-11-08
申请号：US11418421
申请日：2006-05-03
CPC分类号： A01N1/02 , A01N1/0221
摘要： A cryopreservation medium and a cryopreservation method which make it possible to cryopreserve tissues and cells with high viability are provided. A cryopreservation medium contains polyphenol at a concentration of from 30 to 120 ppm. In an embodiment, a cryopreservation method for tissues and cells comprises: incubating tissues or cells for 2 hours in culture medium supplemented with polyphenol at concentrations of from 30 to 120 ppm; suspending tissues or cells in a cryopreservation medium; freezing the suspension of the tissues or cells in the cryopreservation medium; and storing the frozen suspension of tissues or cells in the cryopreservation medium.
摘要翻译： 提供了一种冷冻保存培养基和一种低温保存方法，其可以以高活力冷冻保存组织和细胞。 冷冻保存培养基含有浓度为30至120ppm的多酚。 在一个实施方案中，用于组织和细胞的冷冻保存方法包括：在补充有30至120ppm浓度的多酚的培养基中孵育组织或细胞2小时; 将组织或细胞悬浮在冷冻保存培养基中; 冻结冷冻保存培养基中组织或细胞的悬浮液; 并将冷冻的组织或细胞悬浮液储存在冷冻保存培养基中。
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公开(公告)号：US20070259329A1
公开(公告)日：2007-11-08
申请号：US11418422
申请日：2006-05-03
IPC分类号： A01N1/02
CPC分类号： A01N1/02 , A01N1/0226
摘要： The invention is directed to an anergy-inducing cellular composition comprising: cells having antigenic determinants on the cell surface and an anergy-inducing compound [e.g., (−)-epigallocatechin-3-o-gallate (EGCG)] attached by its chemical affinity to the antigenic determinants. In one embodiment, the anergy-inducing compound blocks co-stimulatory molecules among the antigenic determinants of the cells, which upon encountering with T-cell renders the latter anergic, that is, unresponsive toward alloantigens. In another embodiment, EGCG is of high purity for its efficient attachment to the antigenic determinants. The anergy-inducing cellular composition is prepared by immersing the cells in a culture media solution (RPMI 1640) containing 50-500 ppm EGCG at low physiological temperatures for one to two hours to minimize the cells' mortality. A specific use of the resulting anergic cellular composition is suggested, which is the attenuation of two undesirable acute allogenic responses resulting from major histocompatibility disparity, namely, transplant rejection and GVHD.
摘要翻译： 本发明涉及一种免疫诱导细胞组合物，其包含：在细胞表面具有抗原决定簇的细胞和通过其化学亲合力附着的无能力诱导化合物[例如（ - ） - 表没食子儿茶素-3-邻氯乙酸酯（EGCG）] 对抗原决定簇。 在一个实施方案中，所述无反应诱导化合物阻断所述细胞的抗原决定簇之间的共刺激分子，其在与T细胞相遇时使得后者不敏感，即对同种异体抗原无反应。 在另一个实施方案中，EGCG与抗原决定簇的有效连接具有高纯度。 通过将细胞浸入含有50-500ppm EGCG的培养基溶液（RPMI 1640）中在低生理温度下浸泡1至2小时来制备无能诱导细胞组合物，以使细胞的死亡率最小化。 提出了所得到的无菌细胞组合物的具体用途，其是由主要组织相容性差异（即移植排斥和GVHD）引起的两种不希望的急性同种异体反应的减弱。
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