National Institute of Advanced Industrial Science and Technology

Wako Pure Chemical Industries, Inc.

Yasuko Onuma

Jun Hirabayashi

Makoto Asashima

Hiroaki Tateno

Yuzuru Ito

Masakazu Fukuda

Masaki Warashina

Atsushi Kuno

An object of the present invention is to provide a method for introducing a target substance into undifferentiated cells, and a carrier therefor, whereby the target substance can be specifically introduced into the undifferentiated cells by contacting the undifferentiated cells with an rBC2LCN-target substance fusion product in which rBC2LCN lectin is fused to the target substance. Particularly, an rBC2LCN-toxin fusion product in which rBC2LCN lectin is fused to a toxin functioning in cells or its domain having the ability to kill cells functions as an agent for eliminating undifferentiated stem cells and can be administered into a medium after inducing the differentiation of the stem cells to reliably kill only the stem cells in an undifferentiated state.

Method of introducing a toxin or dye into mammalian pluripotent cells using rBC2LCN

An object of the present invention is to provide a method for evaluating a differentiation status of cells using a culture supernatant of stem cells. Provided is 'undifferentiation sugar chain marker' composed of the sugar chain structure 'Fuc&#x3b1;1-2Gal&#x3b2;1-3GlcNAc' or &#x201c;Fuc&#x3b1;1-2Gal&#x3b2;1-3GalNAc&#x201d; and capable of sensitively determining the undifferentiated state of stem cells using a culture supernatant of stem cells. Also found is BC2LCN lectin or a modified product thereof capable of sensitively recognizing the &#x201c;undifferentiation sugar chain marker&#x201d; as excellent &#x201c;probe for detecting the undifferentiation sugar chain marker&#x201d; capable of determining the undifferentiation status of cells using a culture supernatant.

Undifferentiated cell detection method and complex carbohydrate detection method

Provided are a method for accurately evaluating the differentiation status of stem cells by selectively staining only stem cells in an undifferentiated state, and a method for positively isolating only stem cells in an undifferentiated state. Specifically provided is a method for determining differentiation of a cell comprising a step of contacting a test cell with a probe comprising protein (A) or (B) below and a step of detecting the presence of binding of the probe to the test cell. The method for determining differentiation of a cell is capable of detecting the presence or absence of an undifferentiated stem cell in test cells by using a probe that specifically reacts with undifferentiated stem cells and detecting the presence of bonding to the test cell. (A) A protein comprising an amino acid sequence shown in SEQ ID NO: 1 and recognizing a sugar chain structure of 'Fuc&#x3b1;1-2Gal&#x3b2;1-3GlcNAc' and/or 'Fuc&#x3b1;1-2Gal&#x3b2;1-3GalNAc;' and (B) a protein comprising an amino acid sequence showing 80% or more similarity to the amino acid sequence shown in SEQ ID NO: 1 and recognizing a sugar chain structure of &#x201c;Fuc&#x3b1;1-2Gal&#x3b2;1-3GlcNAc&#x201d; and/or &#x201c;Fuc&#x3b1;1-2Gal&#x3b2;1-3GalNAc.&#x201d;

Cell differentiation assay method, cell isolation method, method for producing induced pluripotent stem cells, and method for producing differentiated cells

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Ibaraki, Japan

Yasuko Onuma