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1. 发明授权

US10240124B2 Cell culture substrate comprising a laminin and a cadherin 有权
公开(公告)号：US10240124B2
公开(公告)日：2019-03-26
申请号：US14360040
申请日：2012-09-19
申请人： BioLamina AB
发明人： Karl Tryggvason , Sergey Rodin
IPC分类号： C12N5/074 , C07K14/705 , C07K14/78 , C12M1/12 , C12N5/00 , C12N5/0735
摘要： The present disclosure related to isolated laminin-521, methods for making recombinant laminin-521, host cells that express recombinant laminin-521, and compositions containing laminin-521. Laminin-521 can maintain stem cells in vitro pluripotency, enable self-renewal, and enable single cell survival of human embryonic stem cells. When pluripotent human embryonic stem cells are cultured on plates coated with recombinant laminin-521 (laminin-11), in the absence of differentiation inhibitors or feeder cells, the embryonic stem cells proliferate and maintain their pluripotency. It has also been discovered that human recombinant laminin-521 (laminin-11) provides single cell survival of stem cells after complete dissociation into a single cell suspension. Useful cell culture mediums containing at most 3.9 ng/ml of beta fibroblast growth factor (bFGF) are also described herein.

2. 发明申请

US20120156254A1 RECOMBINANT LAMININ-521 有权
标题翻译：重组LAMININ-521
公开(公告)号：US20120156254A1
公开(公告)日：2012-06-21
申请号：US13328145
申请日：2011-12-16
申请人： KARL TRYGGVASON , SERGEY RODIN
发明人： KARL TRYGGVASON , SERGEY RODIN
IPC分类号： B32B1/08 , A61K38/14 , A61K38/18 , C12M3/00 , C12P21/00 , C12N15/63 , A61K9/00 , C07K14/47 , C12N5/071
CPC分类号： C07K14/78 , C12N5/0606 , C12N5/0696 , C12N2501/602 , C12N2501/603 , C12N2501/605 , C12N2501/608 , C12N2510/00 , C12N2533/52 , Y10T428/13
摘要： The present disclosure related to isolated laminin-521, methods for making recombinant laminin-521, host cells that express recombinant laminin-521, and compositions containing laminin-521. Laminin-521 can maintain stem cells in vitro pluripotency, enable self-renewal, and enable single cell survival of human embryonic stem cells. When pluripotent human embryonic stem cells are cultured on plates coated with a matrix of recombinant laminin-521 (laminin 11), in the absence of differentiation inhibitors or feeder cells, the embryonic stem cells proliferate and maintain their pluripotency. It has also been discovered that human recombinant laminin-521 (laminin-11) provides single cell survival of stem cells after complete dissociation into a single cell suspension. Useful cell culture mediums containing at most 3.9 ng/ml of beta fibroblast growth factor (bFGF) are also described herein.
摘要翻译：本公开涉及分离的层粘连蛋白-521，制备重组层粘连蛋白521的方法，表达重组层粘连蛋白-521的宿主细胞和含有层粘连蛋白-51的组合物。层粘连蛋白-521可以维持干细胞的体外多能性，实现自我更新，并能使人类胚胎干细胞的单细胞存活。当多能人胚胎干细胞在涂有重组层粘连蛋白-51（层粘连蛋白11）基质的平板上培养时，在不存在分化抑制剂或饲养细胞的情况下，胚胎干细胞增殖并保持其多能性。还已经发现人重组层粘连蛋白-51（层粘连蛋白-11）在完全解离成单细胞悬液后提供干细胞的单细胞存活。含有至多3.9ng / ml的β成纤维细胞生长因子（bFGF）的有用细胞培养基也在本文中描述。

3. 发明公开

US20240132838A1 METHODS OF PRODUCING RPE CELLS 审中-公开
公开(公告)号：US20240132838A1
公开(公告)日：2024-04-25
申请号：US18453712
申请日：2023-08-21
申请人： BioLamina AB
发明人： Outi Hovatta , Karl Tryggvason
IPC分类号： C12N5/079 , A61K9/00 , A61K35/30 , C12N5/00
CPC分类号： C12N5/0621 , A61K9/0019 , A61K9/0048 , A61K35/30 , C12N5/0056 , C12N2500/92 , C12N2500/98 , C12N2501/115 , C12N2506/02 , C12N2533/50 , C12N2533/52
摘要： The present disclosure relates to the use of laminin-521 in obtaining retinal pigment epithelium (RPE) cells. Pluripotent human embryonic stem cells are cultured on plates coated with recombinant laminin-521 (laminin-11), in totally defined and xeno-free conditions. A first cell culture medium contains a growth factor, and a second cell culture medium does not contain growth factor. The stem cells are first exposed to the first cell culture medium, then exposed to the second cell culture medium for a longer time period. After a number of weeks, clinical grade RPE cells are obtained from the stem cells.

4. 发明申请

US20210261918A1 METHODS FOR PRODUCING RETINAL PIGMENT EPITHELIUM CELLS 有权
公开(公告)号：US20210261918A1
公开(公告)日：2021-08-26
申请号：US17252789
申请日：2019-06-19
申请人： BioLamina AB
发明人： Karl Anders Fredrik Lanner , Álvaro Plaza Reyes , Sandra Petrus Reurer , Sara Padrell Sánchez
IPC分类号： C12N5/079
摘要： Method comprises providing a culture of human pluripotent stem cells adherent on a first substrate comprising a first laminin; exposing the stem cells to a differentiation medium for a first time period of 15 days to 50 days to obtain a first population of adherent cells comprising RPE cells and/or progenitors thereof; at the end of the first time period, dissociating the first population from the first substrate; replating the dissociated first population of cells on a second substrate comprising a second laminin; and culturing the replated first population of cells on the second substrate for a second time period to obtain an expanded and matured second population of cells comprising the RPE cells. The first and second laminins may be independently selected from LN-521, LN-511, LN-111 and LN-121, and are an intact protein or protein fragment. Cell surface markers useful for in vitro generation of RPE cells selected from CD140b, CD56, CD104, CD164, CD220, EGER, GD2, CD184, CD10, CD30, CD49a, CD49b, CD50, CD171, TRA-1-60 and CD326, preferably CD140b, CD56, GD2 and/or CD184. Also included are RPE cells so produced as well as materials and compositions utilizing such RPE cells for various treatments.

5. 发明授权

US11001807B2 Differentiation of pluripotent stem cells and cardiac progenitor cells into striated cardiomyocyte fibers using laminins LN-511, LN-521 and LN-221 有权
公开(公告)号：US11001807B2
公开(公告)日：2021-05-11
申请号：US14895669
申请日：2014-07-02
申请人： National University of Singapore , BioLamina AB
发明人： Karl Tryggvason , Yan Wen Yap , Sun Yi , Kristian Tryggvason
IPC分类号： C12N5/077 , C12Q1/6876
摘要： The present disclosure describes methods of differentiating cardiomyocyte progenitor cells and mature cardiomyocyte cells from pluripotent stem cells. The methods may include differentiating pluripotent stems cells on a substrate including (i) laminin-511 or 521 and (ii) laminin 221. The mature cardiomyocyte cells produced by the method may form a human heart muscle cell line for use in regenerative cardiology.

6. 发明授权

US09499794B2 Maintenance of differentiated cells with laminins 有权
标题翻译：维持层粘连蛋白分化细胞
公开(公告)号：US09499794B2
公开(公告)日：2016-11-22
申请号：US13866177
申请日：2013-04-19
申请人： Karl Tryggvason , Karl Kristian Tryggvason , Anna Domogatskaya
发明人： Karl Tryggvason , Karl Kristian Tryggvason , Anna Domogatskaya
IPC分类号： C12N5/071 , C12N5/00
CPC分类号： C12N5/0068 , C12N5/0676 , C12N2501/998 , C12N2533/52
摘要： The present disclosure describes methods of maintaining the phenotype of differentiated cells. Generally, the natural environment of the body is replicated for the differentiated cell. The differentiated cell is plated on a cell culture substrate comprising a laminin, such as laminin-521 or laminin-511. The substrate may also contain a cadherin. This maintains the phenotype of the differentiated cell.
摘要翻译：本公开描述了维持分化细胞表型的方法。通常，体内的天然环境被复制用于分化细胞。将分化的细胞接种在包含层粘连蛋白如层粘连蛋白-51或层粘连蛋白-511的细胞培养基质上。底物也可以含有钙粘着素。这维持了分化细胞的表型。

7. 发明申请

US20130280750A1 MAINTENANCE OF DIFFERENTIATED CELLS WITH LAMININS 有权
标题翻译：维持细胞与层间粘附
公开(公告)号：US20130280750A1
公开(公告)日：2013-10-24
申请号：US13866177
申请日：2013-04-19
申请人： Karl Tryggvason , Karl Kristian Tryggvason , Anna Domogatskaya
发明人： Karl Tryggvason , Karl Kristian Tryggvason , Anna Domogatskaya
IPC分类号： C12N5/071
CPC分类号： C12N5/0068 , C12N5/0676 , C12N2501/998 , C12N2533/52
摘要： The present disclosure describes methods of maintaining the phenotype of differentiated cells. Generally, the natural environment of the body is replicated for the differentiated cell. The differentiated cell is plated on a cell culture substrate comprising a laminin, such as laminin-521 or laminin-511. The substrate may also contain a cadherin. This maintains the phenotype of the differentiated cell.
摘要翻译：本公开描述了维持分化细胞表型的方法。通常，体内的天然环境被复制用于分化细胞。将分化的细胞接种在包含层粘连蛋白如层粘连蛋白-51或层粘连蛋白-511的细胞培养基质上。底物也可以含有钙粘着素。这维持了分化细胞的表型。

8. 发明申请

US20210301257A1 DIFFERENTIATION OF PLURIPOTENT STEM CELLS AND CARDIAC PROGENITOR CELLS INTO STRIATED CARDIOMYOCYTE FIBERS USING LAMININS LN-511, LN-521 AND LN-221 有权
公开(公告)号：US20210301257A1
公开(公告)日：2021-09-30
申请号：US17226611
申请日：2021-04-09
申请人： National University of Singapore , BioLamina AB
发明人： Karl Tryggvason , Yan Wen Yap , Sun Yi , Kristian Tryggvason
IPC分类号： C12N5/077 , C12Q1/6876
摘要： The present disclosure describes methods of differentiating cardiomyocyte progenitor cells and mature cardiomyocyte cells from pluripotent stem cells. The methods may include differentiating pluripotent stems cells on a substrate including (i) laminin-511 or 521 and (ii) laminin 221. The mature cardiomyocyte cells produced by the method may form a human heart muscle cell line for use in regenerative cardiology.

9. 发明申请

US20210139848A1 METHODS OF PRODUCING RPE CELLS 有权
公开(公告)号：US20210139848A1
公开(公告)日：2021-05-13
申请号：US17138327
申请日：2020-12-30
申请人： BioLamina AB
发明人： Outi Hovatta , Karl Tryggvason
IPC分类号： C12N5/079 , C12N5/00 , A61K9/00 , A61K35/30
摘要： The present disclosure relates to the use of laminin-521 in obtaining retinal pigment epithelium (RPE) cells. Pluripotent human embryonic stem cells are cultured on plates coated with recombinant laminin-521 (laminin-11), in totally defined and xeno-free conditions. A first cell culture medium contains a growth factor, and a second cell culture medium does not contain growth factor. The stem cells are first exposed to the first cell culture medium, then exposed to the second cell culture medium for a longer time period. After a number of weeks, clinical grade RPE cells are obtained from the stem cells.

10. 发明授权

US10138292B2 Cell culture medium 有权
公开(公告)号：US10138292B2
公开(公告)日：2018-11-27
申请号：US13328156
申请日：2011-12-16
申请人： Karl Tryggvason , Sergey Rodin
发明人： Karl Tryggvason , Sergey Rodin
IPC分类号： C07K14/78 , C12N5/0735 , C12N5/074
摘要： The present disclosure related to isolated laminin-521, methods for making recombinant laminin-521, host cells that express recombinant laminin-521, and compositions containing laminin-521. Laminin-521 can maintain stem cells in vitro pluripotency, enable self-renewal, and enable single cell survival of human embryonic stem cells. When pluripotent human embryonic stem cells are cultured on plates coated with recombinant laminin-521 (laminin-11), in the absence of differentiation inhibitors or feeder cells, the embryonic stem cells proliferate and maintain their pluripotency. It has also been discovered that human recombinant laminin-521 (laminin-11) provides single cell survival of stem cells after complete dissociation into a single cell suspension. Useful cell culture mediums containing at most 3.9 ng/ml of beta fibroblast growth factor (bFGF) are also described herein.

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