α6β1- and αV-integrins are required for long-term self-renewal of murine embryonic stem cells in the absence of LIF

BMC Cell Biology

Aki Manninen¹  Jayendrakishore Tanjore Ramanathan²  Riitta Palovuori¹  Sandhanakrishnan Cattavarayane¹ 

[1] Biocenter Oulu, Oulu Center for Cell-Matrix Research, Faculty of Biochemistry and Molecular Medicine, University of Oulu, Aapistie 5, Oulu 90220, Finland;Current address: Université de Lorraine, CS 25233, Nancy cedex, 54052, France

关键词: Adhesion;    Self-renewal;    Extracellular matrix;    Integrin;    Embryonic stem cells;   

Others  :  1137096
DOI  :  10.1186/s12860-015-0051-y

【 摘 要 】

Background

The growth properties and self-renewal capacity of embryonic stem (ES) cells are regulated by their immediate microenvironment such as the extracellular matrix (ECM). Integrins, a central family of cellular ECM receptors, have been implicated in these processes but their specific role in ES cell self-renewal remains unclear.

Results

Here we have studied the effects of different ECM substrates and integrins in mouse ES cells in the absence of Leukemia Inhibitory Factor (LIF) using short-term assays as well as long-term cultures. Removal of LIF from ES cell culture medium induced morphological differentiation of ES cells into polarized epistem cell-like cells. These cells maintained epithelial morphology and expression of key stemness markers for at least 10 passages in the absence of LIF when cultured on laminin, fibronectin or collagen IV substrates. The specific functional roles of α6-, αV- and β1-integrin subunits were dissected using stable lentivirus-mediated RNAi methodology. β1-integrins were required for ES cell survival in long-term cultures and for the maintenance of stem cell marker expression. Inhibition of α6-integrin expression compromised self-renewal on collagen while αV-integrins were required for robust ES cell adhesion on laminin. Analysis of the stemness marker expression revealed subtle differences between α6- and αV-depleted ES cells but the expression of both was required for optimal self-renewal in long-term ES cell cultures.

Conclusions

In the absence of LIF, long-term ES cell cultures adapt an epistem cell-like epithelial phenotype and retain the expression of multiple stem cell markers. Long-term maintenance of such self-renewing cultures depends on the expression of β1-, α6- and αV-integrins.

【 授权许可】

   
2015 Cattavarayane et al.; licensee BioMed Central.

【 图 表 】

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