|Cancer||Cell death||Cell cycle||Cytoskeleton||Exo/endocytosis||Differentiation||Division||Organelles||Signalling||Stem cells||Trafficking|
Replacement of mouse embryonic fibroblasts with bone marrow stromal cells for use in establishing and maintaining embryonic stem cells in mice
Chae Hyun Lee*, Jun Hong Park*, Jae Hee Lee†, Ji Yeon Ahn†, Jong Heum Park*, Bo‑Ram Lee‡, Dae Yong Kim‡ and Jeong Mook Lim*†1
*Department of Agricultural Biotechnology, Seoul National University, Seoul 151921, Korea, †Laboratory of Stem Cell and Bioevaluation, WCU Biomodulation Program, Seoul National University, Seoul 151742, Korea, and ‡Department of Veterinary Pathology, College of Veterinary Medicine, Seoul National University, Seoul 151742, Korea
We have investigated the use of BMSC (bone marrow stromal cell) as a feeder cell for improving culture efficiency of ESC (embryonic stem cell). B6CBAF1 blastocysts or ESC stored after their establishment were seeded on to a feeder layer of either SCA-1+/CD45−/CD11b− BMSC or MEF (mouse embryonic fibroblast). Feeder cell activity in promoting ESC establishment from the blastocysts and in supporting ESC maintenance did not differ significantly between BMSC and MEF feeders. However, the highest efficiency of colony formation after culturing of inner cell mass cells of blastocysts was observed with the BMSC line that secreted the largest amount of LIF (leukaemia inhibitory factor). Exogenous LIF was essential for the ESC establishment on BMSC feeder, but not for ESC maintenance. Neither change in stem cell-specific gene expression nor increase in stem cell aneuploidy was detected after the use of BMSC feeder. We conclude that BMSC can be utilized as the feeder of ESC, which improves culture efficiency.
Key words: bone marrow stromal cell (BMSC), embryonic stem cell (ESC), feeder cell
Abbreviations: BMSC, bone marrow stromal cell, DMEM, Dulbecco's modified Eagle's medium, EB, embryoid body, ESC, embryonic stem cell, FBS, fetal bovine serum, ICM, inner cell mass, JAK, Janus kinase, LIF, leukaemia inhibitory factor, MEF, mouse embryonic fibroblast, NOD, non-obese diabetic, PI, propidium iodide, SCID, severe combined immunodeficiency, STAT, signal transducer and activator of transcription
1To whom correspondence should be addressed (email email@example.com).
Received 22 July 2011/28 November 2011; accepted 6 February 2012
Published as Cell Biology International Immediate Publication 6 February 2012, doi:10.1042/CBI20110395
© The Author(s) Journal compilation © 2012 International Federation for Cell Biology