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 tidsskriftTidsskriftartikelfagfællebedømt

Harvard

Hey, CAB, Saltõkova, KB, Bisgaard, HC & Møller, LB 2018, 'Comparison of two different culture conditions for derivation of early hiPSC', Cell Biology International, bind 42, nr. 11, s. 1467-1473. https://doi.org/10.1002/cbin.10966

APA

Hey, C. A. B., Saltõkova, K. B., Bisgaard, H. C., & Møller, L. B. (2018). Comparison of two different culture conditions for derivation of early hiPSC. Cell Biology International, 42(11), 1467-1473. https://doi.org/10.1002/cbin.10966

Vancouver

Hey CAB, Saltõkova KB, Bisgaard HC, Møller LB. Comparison of two different culture conditions for derivation of early hiPSC. Cell Biology International. 2018;42(11):1467-1473. https://doi.org/10.1002/cbin.10966

Author

Hey, Caroline A.B. ; Saltõkova, Katarina B. ; Bisgaard, Hanne C. ; Møller, Lisbeth B. / Comparison of two different culture conditions for derivation of early hiPSC. I: Cell Biology International. 2018 ; Bind 42, Nr. 11. s. 1467-1473.

Bibtex

@article{14f23a154f194c12b4b0429deaab688d,
title = "Comparison of two different culture conditions for derivation of early hiPSC",
abstract = "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.",
keywords = "E8, iPSC, Matrigel, mTeSR1, Vitronectin",
author = "Hey, {Caroline A.B.} and Salt{\~o}kova, {Katarina B.} and Bisgaard, {Hanne C.} and M{\o}ller, {Lisbeth B.}",
year = "2018",
doi = "10.1002/cbin.10966",
language = "English",
volume = "42",
pages = "1467--1473",
journal = "Cell Biology International",
issn = "1065-6995",
publisher = "Academic Press",
number = "11",

}

RIS

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