TY - JOUR
T1 - Dj-1 can replace fgf-2 for long-term culture of human pluripotent stem cells in defined media and feeder-free condition
AU - Kim, Julee
AU - Baek, Sangki
AU - Hong, Yean Ju
AU - de Paula, Michelle Novais
AU - Prima, Musharrat Jahan
AU - Oh, Yeon Mok
AU - Cha, Sun Shin
AU - Do, Jeong Tae
AU - Jang, Yeon Jin
AU - Choe, Han
N1 - Funding Information:
This study was supported by grants (KNRF-2019000772 and NRF-2015K1A4A3046807) from the National Research Foundation of Korea and a grant (2019-IL0307-01) from the Asan Institute for Life Sciences, Seoul, Korea. We thank the electron microscopy core facility at the ConveR-gence mEDIcine research cenTer (CREDIT), Asan Medical Center, Seoul, Korea for support and in-strumentation. S.-S.C. was supported by the project entitled ‘Development of Biomedical materials based on marine proteins’ from the Ministry of Oceans and Fisheries, Korea.
Funding Information:
Funding: This study was supported by grants (KNRF-2019000772 and NRF-2015K1A4A3046807) from the National Research Foundation of Korea and a grant (2019-IL0307-01) from the Asan Institute for Life Sciences, Seoul, Korea. We thank the electron microscopy core facility at the ConveRgence mEDIcine research cenTer (CREDIT), Asan Medical Center, Seoul, Korea for support and instrumentation. S.-S.C. was supported by the project entitled ‘Development of Biomedical materials based on marine proteins’ from the Ministry of Oceans and Fisheries, Korea.
Publisher Copyright:
© 2021 by the authors. Licensee MDPI, Basel, Switzerland.
PY - 2021/6/1
Y1 - 2021/6/1
N2 - Conventional human pluripotent stem cell (hPSC) cultures require high concentrations of expensive human fibroblast growth factor 2 (hFGF-2) for hPSC self-renewal and pluripotency in defined media for long-term culture. The thermal instability of the hFGF-2 mandates media change every day, which makes hPSC culture costly and cumbersome. Human DJ-1 (hDJ-1) can bind to and stimulate FGF receptor-1. In this study, for the first time, we have replaced hFGF-2 with hDJ-1 in the essential eight media and maintained the human embryonic stem cells (hESCs), H9, in the defined media at feeder-free condition. After more than ten passages, H9 in both groups still successfully maintained the typical hESC morphology and high protein levels of pluripotency markers, SSEA4, Tra1-60, Oct4, Nanog, and ALP. DNA microarray revealed that more than 97% of the 21,448 tested genes, including the pluripotency markers, Sox2, Nanog, Klf4, Lin28A, Lin28B, and Myc, have similar mRNA levels between the two groups. Karyotyping revealed no chromosome abnor-malities in both groups. They also differentiated sufficiently into three germ layers by forming in vitro EBs and in vivo teratomas. There were some variations in the RT-qPCR assay of several plu-ripotency markers. The proliferation rates and the mitochondria of both groups were also different. Taken together, we conclude that hDJ-1 can replace hFGF-2 in maintaining the self-renewal and the pluripotency of hESCs in feeder-free conditions.
AB - Conventional human pluripotent stem cell (hPSC) cultures require high concentrations of expensive human fibroblast growth factor 2 (hFGF-2) for hPSC self-renewal and pluripotency in defined media for long-term culture. The thermal instability of the hFGF-2 mandates media change every day, which makes hPSC culture costly and cumbersome. Human DJ-1 (hDJ-1) can bind to and stimulate FGF receptor-1. In this study, for the first time, we have replaced hFGF-2 with hDJ-1 in the essential eight media and maintained the human embryonic stem cells (hESCs), H9, in the defined media at feeder-free condition. After more than ten passages, H9 in both groups still successfully maintained the typical hESC morphology and high protein levels of pluripotency markers, SSEA4, Tra1-60, Oct4, Nanog, and ALP. DNA microarray revealed that more than 97% of the 21,448 tested genes, including the pluripotency markers, Sox2, Nanog, Klf4, Lin28A, Lin28B, and Myc, have similar mRNA levels between the two groups. Karyotyping revealed no chromosome abnor-malities in both groups. They also differentiated sufficiently into three germ layers by forming in vitro EBs and in vivo teratomas. There were some variations in the RT-qPCR assay of several plu-ripotency markers. The proliferation rates and the mitochondria of both groups were also different. Taken together, we conclude that hDJ-1 can replace hFGF-2 in maintaining the self-renewal and the pluripotency of hESCs in feeder-free conditions.
KW - DJ-1
KW - Defined media
KW - FGF-2
KW - Feeder-free
KW - HPSC
UR - http://www.scopus.com/inward/record.url?scp=85106888322&partnerID=8YFLogxK
U2 - 10.3390/ijms22115954
DO - 10.3390/ijms22115954
M3 - Article
C2 - 34073063
AN - SCOPUS:85106888322
SN - 1661-6596
VL - 22
JO - International Journal of Molecular Sciences
JF - International Journal of Molecular Sciences
IS - 11
M1 - 5954
ER -