Composition and method for enabling proliferation of pluripotent stem cells

A technology of pluripotent stem cells and compositions, which can be applied to the composition field of stem cell adhesion, proliferation and self-renewal, and the composition field of stem cells, which can solve problems such as inability to use in clinical practice.

Active Publication Date: 2009-12-16
卡尔·特吕格瓦松 +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, these coatings are of allogeneic origin and thus cannot be used clinically according to FDA regulations (Hovatta, O. and Skottman, H. (2005); Feeder-free derivation of human embryonic stem-cell lines; Lancet 365, 1601-1603 )

Method used

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  • Composition and method for enabling proliferation of pluripotent stem cells
  • Composition and method for enabling proliferation of pluripotent stem cells
  • Composition and method for enabling proliferation of pluripotent stem cells

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Experimental program
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Embodiment Construction

[0026] It has been found that in the presence of the mitogenic factor bFGF (10ng / ml) and the absence of any differentiation inhibitors, when pluripotent mouse embryonic stem cells are coated with recombinant human laminin-5 (laminin- 332) or laminin-10 (laminin-511), the cells proliferate and maintain their pluripotency for at least 140 days (23 passages) ( Figure 1a , Figure 6 ). Expression of pluripotent markers such as Oct4, TERT, Sox2, Nanog and UTF1 ( figure 2 , Figure 3a-3d , Figure 4 ) And the proliferation rate ( Figure 1a ) Also remains stable. In addition, it is noted that the adhesion of embryonic cells to laminin-5 or laminin-10 molecules is related to the latter's ability to support embryonic cell self-renewal ( Figure 5a-5c ).

[0027] In contrast, when mouse embryonic stem cells are coated with mouse (EHS) laminin-1 (laminin-111), laminin-8 (laminin-411), and Matrigel under the same conditions TM , Or when cultured on gelatin plates, the pluripotent cells sto...

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Abstract

The present disclosure is directed to the development of compositions, such as extracellular matrices, and processes for using the same, for culturing stem cells in vitro in an undifferentiated state. In this regard, it has been discovered that when pluripotent mouse and human embryonic stem cells are cultured on plates coated with recombinant laminin-10 (laminin-51 1 ) or laminin-5 (laminin-322), or their functional domains, the embryonic stem cells proliferated and maintained their pluripotency.

Description

[0001] Cross-references to related applications [0002] This application claims the priority of the U.S. Provisional Application Serial No. 60 / 883,406 filed on January 4, 2007, the entire contents of which are hereby incorporated by reference. Background technique [0003] In many exemplary embodiments, the present disclosure generally relates to compositions and methods capable of adhering, proliferating, and maintaining self-renewal of pluripotent or undifferentiated stem cells in vitro. These stem cells include embryonic stem cells (such as murine embryonic stem cells or human embryonic stem cells) and bone marrow stem cells. [0004] Stem cells are undifferentiated cells from which specialized cells are subsequently derived. Embryonic stem cells have extensive self-renewal capabilities and have the pluripotent potential to differentiate into all three germ layer cells. They can be used for therapeutic purposes, and can provide unlimited cell sources for tissue transplantation ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/06C12N5/0735
CPCC12N2533/52C12N5/0606C12N5/00C12N5/06C12N5/0602
Inventor 卡尔·特吕格瓦松安娜·多莫加茨卡娅谢尔盖·罗丁
Owner 卡尔·特吕格瓦松
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