TY - JOUR
T1 - Differentiation of human embryonic stem cells to cardiomyocytes on microcarrier cultures
AU - Ting, Sherwin
AU - Lecina, Marti
AU - Reuveny, Shaul
AU - Oh, Steve
N1 - Sherwin Ting; Martí Lecina; Shaul Reuveny; Steve Oh
PY - 2012/5/1
Y1 - 2012/5/1
N2 - We have developed an improved cardiomyocyte differentiation protocol where we stabilized embryoid bodies (EB) in serum- and insulin-free medium (bSFS) supplemented with p38 MAP kinase inhibitor (SB203580) by addition of 10 μm laminin-coated positively charged (protamine sulfate derivatized TSKgel Tresyl-5PW) microcarriers. This protocol achieved a maximum 3-fold cell expansion, differentiation efficiency of 20%, and an overall cardiomyocyte yield of 3 × 105 CM/ml in static conditions. In comparison, EB cultures achieved 1.5-fold cell expansion, differentiation efficiency of 15%, and an overall cardiomyocyte yield of 1.1 × 105 CM/ml. The scalability of this platform was demonstrated in suspended spinner cultures, producing a maximum of 2.14 × 105 CM/ml in 50-ml cultures. This yield is two-fold higher than the control static EB-based platform (1.1 × 105 CM/ml), and seven-fold higher than yields reported in literature, 3.1-9 × 104 CM/ml. The robustness of this protocol was tested with HES-3 and H1 cell lines.
AB - We have developed an improved cardiomyocyte differentiation protocol where we stabilized embryoid bodies (EB) in serum- and insulin-free medium (bSFS) supplemented with p38 MAP kinase inhibitor (SB203580) by addition of 10 μm laminin-coated positively charged (protamine sulfate derivatized TSKgel Tresyl-5PW) microcarriers. This protocol achieved a maximum 3-fold cell expansion, differentiation efficiency of 20%, and an overall cardiomyocyte yield of 3 × 105 CM/ml in static conditions. In comparison, EB cultures achieved 1.5-fold cell expansion, differentiation efficiency of 15%, and an overall cardiomyocyte yield of 1.1 × 105 CM/ml. The scalability of this platform was demonstrated in suspended spinner cultures, producing a maximum of 2.14 × 105 CM/ml in 50-ml cultures. This yield is two-fold higher than the control static EB-based platform (1.1 × 105 CM/ml), and seven-fold higher than yields reported in literature, 3.1-9 × 104 CM/ml. The robustness of this protocol was tested with HES-3 and H1 cell lines.
KW - Cardiomyocytes;SB203580
KW - Hesc;differentiation
KW - Human embryonic
KW - Microcarriers
KW - Scale up
KW - Stem cells
UR - http://www.scopus.com/inward/record.url?scp=84866033931&partnerID=8YFLogxK
U2 - 10.1002/9780470151808.sc01d07s21
DO - 10.1002/9780470151808.sc01d07s21
M3 - Article
C2 - 22605644
AN - SCOPUS:84866033931
SN - 1941-7322
VL - 21
JO - Current Protocols in Stem Cell Biology
JF - Current Protocols in Stem Cell Biology
IS - 1
M1 - 1D.7
ER -