A novel two-step procedure to expand cardiac Sca-1+ cells clonally.
Resident cardiac stem cells (CSCs) are characterized by their capacity to self-renew in culture, and are multipotent for forming normal cell types in hearts. CSCs were originally isolated directly from enzymatically digested hearts using stem cell markers. However, long exposure to enzymatic digestion can affect the integrity of stem cell markers on the cell surface, and also compromise stem cell function. Alternatively resident CSCs can migrate from tissue explant and form cardiospheres in culture. However, fibroblast contamination can easily occur during CSC culture. To avoid these problems, we developed a two-step procedure by growing the cells before selecting the Sca-1+ cells and culturing in cardiac fibroblast conditioned medium, they avoid fibroblast overgrowth.
Tang
Yao Liang
YL
Stem Cell Biology, Keck Graduate Institute, 535 Watson Dr., Claremont, CA 91711, USA. ytang@kgi.edu
Shen
Leping
L
Qian
Keping
K
Phillips
M Ian
MI
D000818:Animals; D018929:Cell Culture Techniques; D002454:Cell Differentiation; D049109:Cell Proliferation; D002478:Cells, Cultured; D019976:Cloning, Organism / Q000379:methods*; D018920:Coculture Techniques / Q000379:methods*; D005347:Fibroblasts / Q000166:cytology* / Q000502:physiology; D051379:Mice; D032383:Myocytes, Cardiac / Q000166:cytology* / Q000502:physiology; D013234:Stem Cells / Q000166:cytology* / Q000502:physiology; D023822:Tissue Engineering / Q000379:methods