Comparison of two different culture conditions for derivation of early hiPSC
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Comparison of two different culture conditions for derivation of early hiPSC. / Hey, Caroline A.B.; Saltõkova, Katarina B.; Bisgaard, Hanne C.; Møller, Lisbeth B.
I: Cell Biology International, Bind 42, Nr. 11, 2018, s. 1467-1473.Publikation: Bidrag til tidsskrift › Tidsskriftartikel › Forskning › fagfællebedømt
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TY - JOUR
T1 - Comparison of two different culture conditions for derivation of early hiPSC
AU - Hey, Caroline A.B.
AU - Saltõkova, Katarina B.
AU - Bisgaard, Hanne C.
AU - Møller, Lisbeth B.
PY - 2018
Y1 - 2018
N2 - Different culture-systems for derivation of induced pluripotent stem cells (iPSC) in vitro from human fibroblasts have been established. Here, we compared the efficacy of two different feeder-free culture-systems; Matrigel-coated surfaces in combination with mTeSR1 medium versus Vitronectin-coated surfaces in combination with Essential 8 (E8) medium. The comparison was performed by counting the number of emerging iPSC-looking colonies of re-programmed fibroblasts. The fibroblasts were re-programmed using episomal plasmids expressing OCT3/4, SOX2, KLF4, L-MYC, LIN28, and a p53 knock down shP53. Three different fibroblast lines, K40 and K48 from healthy controls and BBS1 from a patient with Bardet–Biedl syndrome, were used in two independent setups. The BBS1 line was used in both setups in combination with K40 and K48 respectively. In all four re-programming experiments, we observed a significantly higher number of emerging colonies with the combination Matrigel/mTeSR1 as compared to the combination Vitronectin/E8. The presence of iPSC was verified by alkaline phosphatase and Tra-1-60 staining. Furthermore, a higher expression of the pluripotency-associated markers NANOG and SOX2 in cells under Matrigel/mTeSR1 conditions compared with Vitronectin/E8 supported the higher proportion of iPSC on Matrigel/mTeSR1 plates. In conclusion, the combination Matrigel/mTeSR1 is more efficient for derivation of iPSC compared to the Vitronectin/E8 combination.
AB - Different culture-systems for derivation of induced pluripotent stem cells (iPSC) in vitro from human fibroblasts have been established. Here, we compared the efficacy of two different feeder-free culture-systems; Matrigel-coated surfaces in combination with mTeSR1 medium versus Vitronectin-coated surfaces in combination with Essential 8 (E8) medium. The comparison was performed by counting the number of emerging iPSC-looking colonies of re-programmed fibroblasts. The fibroblasts were re-programmed using episomal plasmids expressing OCT3/4, SOX2, KLF4, L-MYC, LIN28, and a p53 knock down shP53. Three different fibroblast lines, K40 and K48 from healthy controls and BBS1 from a patient with Bardet–Biedl syndrome, were used in two independent setups. The BBS1 line was used in both setups in combination with K40 and K48 respectively. In all four re-programming experiments, we observed a significantly higher number of emerging colonies with the combination Matrigel/mTeSR1 as compared to the combination Vitronectin/E8. The presence of iPSC was verified by alkaline phosphatase and Tra-1-60 staining. Furthermore, a higher expression of the pluripotency-associated markers NANOG and SOX2 in cells under Matrigel/mTeSR1 conditions compared with Vitronectin/E8 supported the higher proportion of iPSC on Matrigel/mTeSR1 plates. In conclusion, the combination Matrigel/mTeSR1 is more efficient for derivation of iPSC compared to the Vitronectin/E8 combination.
KW - E8
KW - iPSC
KW - Matrigel
KW - mTeSR1
KW - Vitronectin
U2 - 10.1002/cbin.10966
DO - 10.1002/cbin.10966
M3 - Journal article
C2 - 29603519
AN - SCOPUS:85046763630
VL - 42
SP - 1467
EP - 1473
JO - Cell Biology International
JF - Cell Biology International
SN - 1065-6995
IS - 11
ER -
ID: 209546308