Long-term self-renewal of human pluripotent stem cells on human recombinant laminin-511.
We describe a system for culturing human embryonic stem (hES) cells and induced pluripotent stem (iPS) cells on a recombinant form of human laminin-511, a component of the natural hES cell niche. The system is devoid of animal products and feeder cells and contains only one undefined component, human albumin. The hES cells self-renewed with normal karyotype for at least 4 months (20 passages), after which the cells could produce teratomas containing cell lineages of all three germ layers. When plated on laminin-511 in small clumps, hES cells spread out in a monolayer, maintaining cellular homogeneity with approximately 97% OCT4-positive cells. Adhesion of hES cells was dependent on alpha6beta1 integrin. The use of homogeneous monolayer hES or iPS cell cultures provides more controllable conditions for the design of differentiation methods. This xeno-free and feeder-free system may be useful for the development of cell lineages for therapeutic purposes.
Division of Matrix Biology, Department of Medical Biochemistry and Biophysics, Karolinska Institute, Stockholm, Sweden.
Sergey
S
Rodin
Anna
A
Domogatskaya
Susanne
S
Ström
Emil M
EM
Hansson
Kenneth R
KR
Chien
José
J
Inzunza
Outi
O
Hovatta
Karl
K
Tryggvason
D002448:Cell Adhesion / Q000187:drug effects; D018929:Cell Culture Techniques / Q000379:methods*; D002460:Cell Line; D002462:Cell Membrane / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D005434:Flow Cytometry; D006801:Humans; D057026:Induced Pluripotent Stem Cells / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D016023:Integrins / Q000378:metabolism; D007797:Laminin / Q000494:pharmacology*; D039904:Pluripotent Stem Cells / Q000166:cytology*; D011956:Receptors, Cell Surface / Q000378:metabolism; D011994:Recombinant Proteins / Q000494:pharmacology*; D013997:Time Factors
D016023:Integrins; D007797:Laminin; D011956:Receptors, Cell Surface; D011994:Recombinant Proteins; C551997:laminin-511, human