Induced extended pluripotent stem cells, methods of making and using

A pluripotent cell and pluripotency technology, applied in the field of pluripotency compositions in vivo, can solve the problems of human embryonic stem cells hindering operation and development

Active Publication Date: 2018-11-23
BEIHAO STEM CELL & REGENERATIVE MEDICINE RES INST CO LTD +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

As an additional example, poor survival of human embryonic stem cells after cell dissociation hampers further manipulation and development

Method used

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  • Induced extended pluripotent stem cells, methods of making and using
  • Induced extended pluripotent stem cells, methods of making and using
  • Induced extended pluripotent stem cells, methods of making and using

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Experimental program
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Embodiment

[0148] method

[0149] small molecule library

[0150] Small molecule libraries for screening were purchased or generated in-house as described in Table 1.

[0151] Table 1. Small molecule compound libraries used for screening

[0152]

[0153] *This library was generated in-house and includes 108 selected small molecules related to pluripotency or epigenetic modification.

[0154] Chemical Screening Based on Luciferase Reporter Gene Assay

[0155] Initial efforts have focused on identifying new small molecules that support naive pluripotency in humans. Based on 2i plus LIF conditions (ERK inhibitor PD0325901, GSK3β inhibitor CHIR99021, and human LIF (hLIF)) that support naive pluripotency in mice, an initial screen was performed using primed hES cells to identify those capable of activating naive pluripotency Marker OCT4 distal enhancer (DE) compound (Yeom et al., Development, 122:881-894 (1996); Tesar et al., Nature, 448:196-199 (2007) ( Figure 1A and Table 1).

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Abstract

Factors for extending the ability of isolated pluripotent stem cells to generate extraembryonic lineages in vivo, following in vitro culture, herein, chemical extenders of pluripotency (CEP). Methodsof extending the ability of a pluripotent cell to generate embryonic and extraembryonic lineages. The cell to be reprogrammed is contacted with effective amounts of the CEPs for a sufficient period oftime to reprogram the cell into a chemically induced extended pluripotent cell (ciEPSC). ciEPSC are identified as an extended pluripotent cell based on properties including: (i) morphologically and (ii) functionally for example, based on their ability contribute to both TE and ICM, in vivo. The ciEPSCs can be cultured or induced to differentiate into cells of a desired type, and used in a numberof applications, including but not limited to cell therapy and tissue engineering.

Description

field of invention [0001] The present invention relates generally to compositions and methods for extending the in vivo pluripotency of pluripotent stem cells cultured in vitro. Background of the invention [0002] Totipotent cells in the early embryo are the ancestors of all stem cells and are capable of developing into whole organisms, including extraembryonic tissues such as the placenta. Pluripotent cells in the inner cell mass (ICM) are the descendants of totipotent cells and are capable of differentiating into any cell type of the body except extraembryonic tissues. [0003] Animal development is initiated by the fertilization of an egg by a sperm, followed by mitotic cell division, or cleavage, to produce blastomeres. In most animals, the first step in diversifying cell types is the creation of the primordrm, the endoderm, mesoderm, and ectoderm. In general, the endoderm is the precursor of the gastrointestinal tract necessary for nutrient absorption; the mesoderm p...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/00C07K14/54C07K14/715C12N5/07C12N5/074C12N5/0789C12N5/0797C12N5/095C07K16/38C07K16/40
CPCC07K16/38C07K16/40C12N5/0605C12N5/0606C12N2501/15C12N2501/235C12N2501/603C12N2501/999C12N2503/02C12N2510/00C12N15/873A01K2207/12A01K2217/00C12N5/0696A61P25/00A61P35/00A61P43/00A61P3/10A01K67/027A01K67/0271A01K2227/105A61K35/545C12N2501/20C12N2501/2306C12N2501/2311C12N2501/2327C12N2501/2331C12N2501/237
Inventor 邓宏魁杨杨刘蓓徐君
Owner BEIHAO STEM CELL & REGENERATIVE MEDICINE RES INST CO LTD
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