Replacement of mouse embryonic fibroblasts with bone marrow stromal cells for use in establishing and maintaining embryonic stem cells in mice.
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.
Department of Agricultural Biotechnology, Seoul National University, Seoul 151-921, Korea.
Chae Hyun
CH
Lee
Jun Hong
JH
Park
Jae Hee
JH
Lee
Ji Yeon
JY
Ahn
Jong Heum
JH
Park
Bo-Ram
BR
Lee
Dae Yong
DY
Kim
Jeong Mook
JM
Lim
D000818:Animals; D000943:Antigens, Differentiation / Q000235:genetics / Q000378:metabolism; D053624:Blastocyst Inner Cell Mass / Q000166:cytology*; D001854:Bone Marrow Cells / Q000378:metabolism*; D002454:Cell Differentiation; D049109:Cell Proliferation; D018920:Coculture Techniques; D017077:Culture Media, Conditioned / Q000737:chemistry; D053595:Embryonic Stem Cells / Q000502:physiology* / Q000637:transplantation; D061252:Feeder Cells; D005269:Femur / Q000166:cytology; D005347:Fibroblasts / Q000378:metabolism*; D015870:Gene Expression; D059785:Karyotype; D053605:Leukemia Inhibitory Factor / Q000378:metabolism; D051379:Mice; D008813:Mice, Inbred ICR; D016688:Mice, Inbred NOD; D016513:Mice, SCID; D017154:Stromal Cells / Q000378:metabolism*; D013977:Tibia / Q000166:cytology
D000943:Antigens, Differentiation; D017077:Culture Media, Conditioned; D053605:Leukemia Inhibitory Factor