公开(公告)号：US20120178159A1

公开(公告)日：2012-07-12

申请号：US13323475

申请日：2011-12-12

申请人： TENG MA ,  Junho Kim

发明人： TENG MA ,  Junho Kim

IPC分类号： C12N5/077 ,  C12N5/02 ,  C12N5/0775

CPC分类号： A61L27/3804 ,  A61L27/3895 ,  C12N5/0062 ,  C12N5/0663 ,  C12N2500/02 ,  C12N2533/90 ,  C12N2539/10

摘要： The subject invention concerns materials and methods for growing and expanding MSC while maintaining their undifferentiated phenotype, self-renewal ability, and/or multi-lineage potential. In one embodiment, a method of the invention comprises i) seeding freshly isolated MSC on a planar surface or a 3-D scaffold and growing the cells under physiological or low O2 tension for a period of time sufficient to support formation of 3-D ECM network; ii) decellularizing the planar surface or 3-D scaffold; and iii) reseeding the decellularized planar surface or 3-D scaffold with MSCs, whereby the reseeded MSCs can be grown on the scaffold and maintain an undifferentiated phenotype. In one embodiment, the 3-D scaffold comprises or is composed of PET. In one embodiment, the MSC are human MSC (hMSC).

摘要翻译： 本发明涉及用于生长和扩增MSC同时保持其未分化表型，自我更新能力和/或多谱系潜力的材料和方法。 在一个实施方案中，本发明的方法包括：i）在平坦表面或3-D支架上接种新鲜分离的MSC，并在生理或低O 2张力下培养细胞足以支持3-D ECM形成的时间 网络; ii）使平面表面或3-D支架脱细胞; 和iii）用MSCs再剥离脱细胞的平面或3-D支架，由此可将重新植入的MSC植入支架上并保持未分化的表型。 在一个实施方案中，3-D支架包含PET或由PET组成。 在一个实施例中，MSC是人MSC（hMSC）。

公开(公告)号：US20120020934A1

公开(公告)日：2012-01-26

申请号：US13184357

申请日：2011-07-15

申请人： TENG MA

发明人： TENG MA

IPC分类号： A61K35/12 ,  C12N5/0775 ,  A61P19/00 ,  C12N5/071

CPC分类号： C12N5/0068 ,  A61K2035/124 ,  A61L27/12 ,  A61L27/26 ,  A61L27/3834 ,  A61L27/3847 ,  C12N5/0663 ,  C12N2533/18 ,  C12N2533/54 ,  C12N2533/72 ,  C08L5/08 ,  C08L89/06

摘要： The subject invention concern materials and methods for cryopreservation of HCG-cell constructs. In one embodiment, porous HCG scaffolds are provided in a perfusion bioreactor having perfusion chambers that can contain the HCG scaffolds, cells are then seeded in the HCG scaffolds in the perfusion bioreactor, cell culture media is perfused through and the bioreactor operated so as to allow for cell seeding and growth in the HCG scaffold. After a suitable period of time, the cell culture media is removed and the HCG containing cells (HCG-cell constructs) can be washed with a suitable buffer, such as phosphate-buffered saline (PBS). The HCG-cell constructs are then perfused in the bioreactor with a suitable cryopreservation fluid. The cryopreservant can comprise one or more of the following: DMSO, trehalose, glycerol, ethylene glycol, and serum for cell culture (e.g., fetal bovine serum (FBS)). In one embodiment, the HCG-cell constructs are perfused for a suitable period of time with cryopreservant fluid using transverse flow of the fluid in the bioreactor at a suitable flow rate. The HCG-cell constructs (or the perfusion chambers containing them) are then removed from the bioreactor and placed in a cryopreservant media and maintained at increasingly colder temperatures until temperatures reach about −80° C. The frozen HCG-cell constructs (or the chambers containing them) can then be stored at a suitable cryogenic temperature (e.g., in liquid nitrogen) until needed. When needed, frozen HCG-cell constructs can be removed from cold storage and thawed using suitable means (e.g., 37° C. water bath). Cells contemplated for use in the present invention include stem cells, such mesenchymal stem cells. Cells can be animal cells, such as mammalian cells. In one embodiment, the cells are human cells.

摘要翻译： 本发明涉及用于冷冻保存HCG细胞构建体的材料和方法。 在一个实施方案中，在具有可含有HCG支架的灌注室的灌注生物反应器中提供多孔HCG支架，然后将细胞接种在灌注生物反应器中的HCG支架中，通过灌注细胞培养基并且使生物反应器运行以允许 用于HCG支架中的细胞接种和生长。 在合适的时间段之后，除去细胞培养基，并用合适的缓冲液如磷酸盐缓冲盐水（PBS）洗涤含有HCG的细胞（HCG细胞构建体）。 然后将HCG细胞构建体用合适的冷冻保存液在生物反应器中灌注。 冷冻保存剂可以包含以下一种或多种：DMSO，海藻糖，甘油，乙二醇和用于细胞培养的血清（例如胎牛血清（FBS））。 在一个实施方案中，使用流体在生物反应器中的流体以合适的流速将冷冻保存的流体灌注HCG细胞构建体适合的时间。 然后将HCG细胞构建体（或含有它们的灌注室）从生物反应器中移出并置于冷冻保存的培养基中并保持在越来越冷的温度，直到温度达到约-80℃。冷冻的HCG细胞构建体（或室 含有它们）然后可以在合适的低温（例如，在液氮中）储存直至需要。 当需要时，可以使用合适的方法（例如，37℃水浴）将冷冻的HCG-细胞构建体从冷藏中移出并解冻。 考虑用于本发明的细胞包括干细胞，这种间充质干细胞。 细胞可以是动物细胞，例如哺乳动物细胞。 在一个实施方案中，细胞是人细胞。

公开(公告)号：US20110229970A1

公开(公告)日：2011-09-22

申请号：US13041688

申请日：2011-03-07

申请人： TENG MA

发明人： TENG MA

IPC分类号： C12N15/85 ,  C12N5/071 ,  C12N5/0775 ,  C12M3/00 ,  B82Y5/00

CPC分类号： C12M29/10 ,  A61L27/26 ,  A61L27/38 ,  A61L27/46 ,  A61L27/52 ,  A61L27/56 ,  A61L2300/258 ,  A61L2300/624 ,  A61L2430/02 ,  C12M25/14 ,  C12N5/0654 ,  C12N2533/18 ,  C12N2533/54 ,  C12N2533/72 ,  C08L89/06 ,  C08L5/08

摘要： The subject invention concerns a perfusion bioreactor device and methods of using the same. A bioreactor device of the invention can be used to grow cells and tissue in a controlled in vitro environment. A perfusion bioreactor device of the invention can have multiple perfusion chambers that can be controlled individually. Transverse or parallel flow of a fluid can be provided to each chamber. Cells can be seeded on a hydrogel and/or 3D scaffold to provide a 3D environment in the bioreactor device where the cells can adhere, proliferate, migrate, secrete growth and/or differentiation factors, and/or undergo differentiation, etc. The subject invention also concerns hydrogels and 3D scaffolds that can be used to grow and/or differentiate cells thereon.

摘要翻译： 本发明涉及灌注生物反应器装置及其使用方法。 本发明的生物反应器装置可用于在受控的体外环境中生长细胞和组织。 本发明的灌注生物反应器装置可以具有可以单独控制的多个灌注室。 可以向每个室提供流体的横向或平行流动。 细胞可以接种在水凝胶和/或3D支架上以在生物反应器装置中提供3D环境，其中细胞可以粘附，增殖，迁移，分泌生长和/或分化因子，和/或进行分化等。本发明 也涉及可用于在其上生长和/或分化细胞的水凝胶和3D支架。