公开(公告)号：US20220253016A1

公开(公告)日：2022-08-11

申请号：US17729267

申请日：2022-04-26

Applicant: FUJIFILM Corporation

Inventor： Yuta MURAKAMI ,  Ryusuke Osaki ,  Sho Onozawa ,  Sohichiro Nakamura

IPC: G03H1/08 ,  C12N5/0735

Abstract: There is provided a method including generating a phase contrast image of an aggregate of a pluripotent stem cell from a hologram in which the aggregates is captured; deriving a phase contrast amount density obtained by dividing a total phase contrast amount which is a value obtained by integrating a phase contrast amount of each of a plurality of pixels that constitute the phase contrast image, by a volume of the aggregate; and sorting the pluripotent stem cell based on the phase contrast amount density.

公开(公告)号：US20210002605A1

公开(公告)日：2021-01-07

申请号：US17025821

申请日：2020-09-18

Applicant: FUJIFILM Corporation

Inventor： Yuta MURAKAMI

IPC: C12N5/00 ,  C12N5/074

Abstract: An object of the present invention is to provide a method for three-dimensionally culturing a pluripotent stem cell, in which the pluripotent stem cell can maintain good proliferative properties even in a case where the frequency of the medium exchange is reduced. According to the present invention, a method for three-dimensionally culturing a pluripotent stem cell, in which a pluripotent stem cell is cultured in a medium that always contains 35 ng/mL or more of basic fibroblast growth factor until a cell concentration reaches 1.4×106 cells/mL or more, is provided.

公开(公告)号：US20160251613A1

公开(公告)日：2016-09-01

申请号：US15064619

申请日：2016-03-09

Applicant: FUJIFILM CORPORATION

Inventor： Yuta MURAKAMI ,  Sanae NOMIYAMA ,  Keita HAGIYA ,  Yuichi YOSHINO ,  Rie HANDO ,  Yoshihide IWAKI ,  Tasuku SASAKI

IPC: C12N5/00 ,  C12N5/074

CPC classification number: C12N5/0068 ,  C07K7/06 ,  C07K14/00 ,  C07K14/78 ,  C12N5/0696 ,  C12N2500/25 ,  C12N2500/32 ,  C12N2500/38 ,  C12N2500/90 ,  C12N2501/119 ,  C12N2501/15 ,  C12N2533/30 ,  C12N2533/50

Abstract: A culture method for pluripotent stem cells includes culturing pluripotent stem cells on a cell culture surface of a support by using a medium in which the concentration of 2-mercaptoethano is equal to or less than 10 μM in the presence of a polypeptide consisting of 40 to 450 amino acid residues, in which the polypeptide includes (1) a first domain including at least one amino acid sequence selected from the group consisting of an amino acid sequence represented by CSYYQSC (SEQ ID NO: 1) and an amino acid sequence represented by RGD and (2) a second domain including (2-i) an amino acid sequence which is represented by PRPSLAKKQRFRHRNRKGYRSQRGHSRGRNQN (SEQ ID NO: 2), (2-ii) an amino acid sequence which shares sequence identity of equal to or higher than 50% with the amino acid sequence represented by SEQ ID NO: 2 and exhibits adsorbability with respect to the cell culture surface of the support, or (2-iii) an amino acid sequence which is formed by the addition, substitution, or deletion of 1 to 30 amino acids in the amino acid sequence represented by SEQ ID NO: 2 and exhibits adsorbability with respect to the cell culture surface of the support.

Abstract translation: 多能干细胞的培养方法包括通过使用其中2-巯基乙醇的浓度等于或小于10μM的培养基，在多肽组成的培养基中培养多能干细胞，所述多肽由40〜 450个氨基酸残基，其中多肽包括（1）包含至少一个选自由CSYYQSC（SEQ ID NO：1）表示的氨基酸序列的氨基酸序列的氨基酸序列的第一结构域和由 RGD和（2）包含（2-i）由PRPSLAKKQRFRHRNRKGYRSQRGHSRGRNQN（SEQ ID NO：2）表示的氨基酸序列的第二结构域，（2-ii）具有等于或高于 50％与SEQ ID NO：2表示的氨基酸序列相比，表现出对载体的细胞培养表面的吸附性，或（2-iii）通过添加，取代或取代形成的氨基酸序列 在SEQ ID NO：2所示的氨基酸序列中为1〜30个氨基酸，并且相对于载体的细胞培养物表面具有吸附性。

公开(公告)号：US20180208902A1

公开(公告)日：2018-07-26

申请号：US15849552

申请日：2017-12-20

Applicant: FUJIFILM CORPORATION

Inventor： Yuta MURAKAMI

IPC: C12N5/074 ,  C12M1/00

CPC classification number: C12N5/0696 ,  C07K14/50 ,  C07K14/78 ,  C12M1/00 ,  C12M3/00 ,  C12M23/20 ,  C12N2501/115

Abstract: Provided are a method for culturing pluripotent stem cells including a culturing process of bringing pluripotent stem cells into contact with a polypeptide which has cell adhesion activity and includes a first domain for binding bFGF and a second domain for adhering to a culture vessel and culturing the pluripotent stem cells, in which bFGF is bound to the first domain, and the second domain is adhered to the culture vessel; a method for culturing pluripotent stem cells including a culturing process of bringing bFGF, which is directly or indirectly adhered to a culture vessel, and a polypeptide, which has cell adhesion activity and is adhered to the culture vessel, into contact with pluripotent stem cells and culturing the pluripotent stem cells; and applications thereof.

公开(公告)号：US20160272945A1

公开(公告)日：2016-09-22

申请号：US15139497

申请日：2016-04-27

Applicant: FUJIFILM Corporation

Inventor： Keita HAGIYA ,  Sanae MORIOKA ,  Yuta MURAKAMI ,  Rie HANDO ,  Kouo SUZUKI ,  Yoshihide IWAKI ,  Tasuku SASAKI

IPC: C12N5/074 ,  C07K14/78

CPC classification number: C12N5/0696 ,  C07K14/78 ,  C12N5/0018 ,  C12N2500/32 ,  C12N2500/38 ,  C12N2501/115 ,  C12N2501/15 ,  C12N2501/33 ,  C12N2501/998

Abstract: A polypeptide composition induces a pluripotent stem cell culturing property, particularly, an excellent cell growth ability. The polypeptide composition contains a predetermined polypeptide including an amino acid sequence of human vitronectin or an amino acid sequence of a predetermined first region derived from human vitronectin. The polypeptide composition includes a multimeric polypeptide, which is composed of two or more monomers held together by intermolecular cross-linking via cysteine residues included in the first region, in an amount equal to or less than 20% by mass of a total mass of polypeptides contained in the composition. A culture method for pluripotent stem cells includes culturing pluripotent stem cells in the presence of the polypeptide composition. Also provided is a culture vessel including a support which has a cell culture surface and the polypeptide contained in the polypeptide composition disposed on the cell culture surface of the support.

Abstract translation: 多肽组合物诱导多能干细胞培养特性，特别是优良的细胞生长能力。 多肽组合物含有包含人玻连蛋白的氨基酸序列或来自人玻连蛋白的预定第一区的氨基酸序列的预定多肽。 多肽组合物包括多聚体多肽，其由两个或更多个通过分子间交联通过包含在第一区域中的半胱氨酸残基保持在一起的单体组成，其量等于或小于20质量％ 包含在组合物中。 多能干细胞的培养方法包括在多肽组合物存在下培养多能干细胞。 还提供了一种培养容器，其包括具有细胞培养表面的载体和多肽组合中所含的多肽，其设置在载体的细胞培养表面上。

公开(公告)号：US20160244728A1

公开(公告)日：2016-08-25

申请号：US15065217

申请日：2016-03-09

Applicant: FUJIFILM Corporation

Inventor： Keita HAGIYA ,  Yuta MURAKAMI ,  Yuichi YOSHINO ,  Sanae NOMIYAMA ,  Rie HANDO ,  Yoshihide IWAKI ,  Tasuku SASAKI

IPC: C12N5/074

CPC classification number: C12N5/0696 ,  C12N2500/38 ,  C12N2510/00 ,  C12N2533/50 ,  C12N2533/52

Abstract: A culture method for pluripotent stem cells includes obtaining a polypeptide-coated culture surface by applying a polypeptide to a cell culture surface of a support, and culturing pluripotent stem cells by seeding the pluripotent stem cells onto the polypeptide-coated culture surface by using a medium in which the content of an ascorbic acid derivative is equal to or greater than 1.5 mmol/L (mM), in which the polypeptide is (a) a polypeptide having an amino acid sequence represented by SEQ ID NO: 1, (b) a polypeptide having an amino acid sequence, which shares identity of equal to or higher than 80% with the amino acid sequence represented by SEQ ID NO: 1, and having culture performance for pluripotent stem cells, or (c) a polypeptide having an amino acid sequence, which is formed by the deletion, substitution, or addition of one amino acid or several amino acids in SEQ ID NO: 1, and having culture performance for pluripotent stem cells.

Abstract translation: 多能干细胞的培养方法包括通过将多肽施用于载体的细胞培养物表面获得多肽包被的培养物表面，并通过使用培养基将多能干细胞接种到多肽包被的培养物表面上来培养多能干细胞 其中抗坏血酸衍生物的含量等于或大于1.5mmol / L（mM），其中多肽是（a）具有SEQ ID NO：1所示氨基酸序列的多肽，（b）a 具有氨基酸序列的多肽，其与SEQ ID NO：1所示的氨基酸序列具有等于或高于80％的同一性，并具有多能干细胞的培养性能，或（c）具有氨基酸的多肽 序列，其通过SEQ ID NO：1中的一个氨基酸或几个氨基酸的缺失，取代或添加而形成，并具有多能干细胞的培养性能。

公开(公告)号：US20150050737A1

公开(公告)日：2015-02-19

申请号：US14528197

申请日：2014-10-30

Applicant: FUJIFILM CORPORATION

Inventor： Yuta MURAKAMI ,  Rie IWATA ,  Yoshihide IWAKI ,  Tasuku SASAKI

IPC: C12N5/074 ,  C07K14/00

CPC classification number: C12N5/0068 ,  C07K14/001 ,  C07K14/47 ,  C07K14/78 ,  C12N5/0607 ,  C12N5/0696 ,  C12N2533/50 ,  C12N2533/52 ,  C12N2537/00

Abstract: A polypeptide including: (1) a first region containing at least one selected from the group consisting of an amino acid sequence represented by CSYYQSC (SEQ ID NO:1) and an amino acid sequence represented by RGD; and (2) a second region containing (2-i) an amino acid sequence represented by PRPSLAKKQRFRHRNRKGYRSQRGHSRGRNQN (SEQ ID NO:2), (2-ii) an amino acid sequence having an identity of not less than 50% to the amino acid sequence represented by SEQ ID NO:2 and having an adsorption ability to a cultivation container, or (2-iii) an amino acid sequence that is the amino acid sequence represented by SEQ ID NO:2 in which from 1 to 30 amino acid residues are added, substituted, or deleted, and has an adsorption ability to a cultivation container, in which the polypeptide includes from 40 to 450 amino acid residues.

Abstract translation: 一种多肽，包括：（1）含有选自由CSYYQSC（SEQ ID NO：1）表示的氨基酸序列和由RGD表示的氨基酸序列组成的组中的至少一种的第一区域; 和（2）含有（2-i）由PRPSLAKKQRFRHRNRKGYRSQRGHSRGRNQN（SEQ ID NO：2）表示的氨基酸序列的第二区域，（2-ii）与氨基酸具有不小于50％的同一性的氨基酸序列 由SEQ ID NO：2表示的具有对培养容器的吸附能力的序列，或（2-iii）作为SEQ ID NO：2所示的氨基酸序列的氨基酸序列，其中含有1〜30个氨基酸残基 被添加，取代或缺失，并且对培养容器具有吸附能力，其中多肽包含40至450个氨基酸残基。

公开(公告)号：US20230066188A1

公开(公告)日：2023-03-02

申请号：US18053373

申请日：2022-11-08

Applicant: FUJIFILM CORPORATION

Inventor： Yuta MURAKAMI ,  Masaya NAGASE ,  Yukihisa NOGUCHI

IPC: C12Q1/68 ,  C12N5/07 ,  C12Q1/04

Abstract: Provided are a biomarker identifying method including (1) to (4) and an application thereof. (1) An evaluation value for each of a plurality of biomarkers is derived based on annotation information imparted to each of biomarkers, and a measurement target biomarker is selected based on the evaluation value. (2) The evaluation data of the measurement target biomarker is acquired from the cell A and/or the culture system, before the start of culture of the cell A and/or during the culture. (3) The evaluation data of the discrimination marker of the B cell is acquired from the cell A and/or the culture system, at the final stage of the culture of the cell A and/or after the end of the culture. (4) At least one biomarker indicating characteristics of the cell A is identified from among the measurement target biomarkers, based on the data obtained from (2) and (3).

公开(公告)号：US20220244243A1

公开(公告)日：2022-08-04

申请号：US17727049

申请日：2022-04-22

Applicant: FUJIFILM Corporation

Inventor： Yuta MURAKAMI ,  Ryusuke OSAKI ,  Sho ONOZAWA ,  Sohichiro NAKAMURA

IPC: G01N33/50 ,  C12N5/074 ,  C12N5/077 ,  G01N15/14

Abstract: There is provided a method including generating a phase contrast image of an aggregate of a pluripotent stem cell from a hologram in which the aggregate is captured; deriving, based on the phase contrast image, an index value indicating a complexity or simplicity of a contour line, the index value being determined according to a relationship between an area of the aggregate and a length of the contour line of the aggregate; and sorting the pluripotent stem cell based on the index value.

公开(公告)号：US20220017872A1

公开(公告)日：2022-01-20

申请号：US17487833

申请日：2021-09-28

Applicant: FUJIFILM Corporation

Inventor： Yuta MURAKAMI ,  Masaya NAGASE

IPC: C12N5/074 ,  C12N5/10

Abstract: An object of the present invention is to provide a producing method for a pluripotent stem cell capable of differentiating into a specific cell. According to the present invention, there is provided a producing method for a pluripotent stem cell capable of differentiating into a specific cell. According to the present invention, there is provided a pluripotent stem cell capable of differentiating into a specific cell. According to the present invention, there is provided a producing method for a differentiated cell, and a differentiated cell. According to the present invention, there is provided a method for quality evaluation of a pluripotent stem cell. Further, according to the present invention, there is provided a method of screening a pluripotent stem cell capable of differentiating into a specific cell.