Directed Differentiation of Human Pluripotent Stem Cells towards Corneal Endothelial-Like Cells under Defined Conditions

被引:15
|
作者
Gronroos, Pyry [1 ]
Ilmarinen, Tanja [1 ]
Skottman, Heli [1 ]
机构
[1] Tampere Univ, Fac Med & Hlth Technol, Arvo Ylpon Katu 34, Tampere 33520, Finland
基金
芬兰科学院;
关键词
corneal endothelial cells; human pluripotent stem cells; neural crest cells; differentiation; small molecule; retinoic acid;
D O I
10.3390/cells10020331
中图分类号
Q2 [细胞生物学];
学科分类号
071009 ; 090102 ;
摘要
The most crucial function of corneal endothelial cells (CEnCs) is to maintain optical transparency by transporting excess fluid out of stroma. Unfortunately, CEnCs are not able to proliferate in vivo in the case of trauma or dystrophy. Visually impaired patients with corneal endothelial deficiencies that are waiting for transplantation due to massive global shortage of cadaveric corneal transplants are in a great need of help. In this study, our goal was to develop a defined, clinically applicable protocol for direct differentiation of CEnCs from human pluripotent stem cells (hPSCs). To produce feeder-free hPSC-CEnCs, we used small molecule induction with transforming growth factor (TGF) beta receptor inhibitor SB431542, GSK-3-specific inhibitor CHIR99021 and retinoic acid to guide differentiation through the neural crest and periocular mesenchyme (POM). Cells were characterized by the morphology and expression of human (h)CEnC markers with immunocytochemistry and RT-qPCR. After one week of induction, we observed the upregulation of POM markers paired-like homeodomain transcription factor 2 (PITX2) and Forkhead box C1 (FOXC1) and polygonal-shaped cells expressing CEnC-associated markers Zona Occludens-1 (ZO-1), sodium-potassium (Na+/K+)-ATPase, CD166, sodium bicarbonate cotransporter 1 (SLC4A4), aquaporin 1 (AQP1) and N-cadherin (NCAD). Furthermore, we showed that retinoic acid induced a dome formation in the cell culture, with a possible indication of fluid transport by the differentiated cells. Thus, we successfully generated CEnC-like cells from hPSCs with a defined, simple and fast differentiation method.
引用
收藏
页码:1 / 15
页数:15
相关论文
共 50 条
  • [31] Laminin 221 fragment is suitable for the differentiation of human induced pluripotent stem cells into brain microvascular endothelial-like cells with robust barrier integrity
    Hiromasa Aoki
    Misaki Yamashita
    Tadahiro Hashita
    Takahiro Iwao
    Tamihide Matsunaga
    [J]. Fluids and Barriers of the CNS, 17
  • [32] Laminin 221 fragment is suitable for the differentiation of human induced pluripotent stem cells into brain microvascular endothelial-like cells with robust barrier integrity
    Aoki, Hiromasa
    Yamashita, Misaki
    Hashita, Tadahiro
    Iwao, Takahiro
    Matsunaga, Tamihide
    [J]. FLUIDS AND BARRIERS OF THE CNS, 2020, 17 (01)
  • [33] Directed differentiation of human induced pluripotent stem cells into functional cholangiocyte-like cells
    Fotios Sampaziotis
    Miguel Cardoso de Brito
    Imbisaat Geti
    Alessandro Bertero
    Nicholas RF Hannan
    Ludovic Vallier
    [J]. Nature Protocols, 2017, 12 : 814 - 827
  • [34] Directed differentiation of human induced pluripotent stem cells into functional cholangiocyte-like cells
    Sampaziotis, Fotios
    de Brito, Miguel Cardoso
    Geti, Imbisaat
    Bertero, Alessandro
    Hannan, Nicholas R. F.
    Vallier, Ludovic
    [J]. NATURE PROTOCOLS, 2017, 12 (04) : 814 - 827
  • [35] Mechanotransductive Differentiation of Hair Follicle Stem Cells Derived from Aged Eyelid Skin into Corneal Endothelial-Like Cells
    Christian Olszewski
    Jessika Maassen
    Rebecca Guenther
    Claudia Skazik-Voogt
    Angela Gutermuth
    [J]. Stem Cell Reviews and Reports, 2022, 18 : 1668 - 1685
  • [36] Directed Differentiation of Human Induced Pluripotent Stem Cells
    Zhang, Su-Chun
    [J]. FASEB JOURNAL, 2011, 25
  • [37] Directed Differentiation of Human Pluripotent Stem Cells to Microglia
    Douvaras, Panagiotis
    Sun, Bruce
    Wang, Minghui
    Kruglikov, Ilya
    Lallos, Gregory
    Zimmer, Matthew
    Terrenoire, Cecile
    Zhang, Bin
    Gandy, Sam
    Schadt, Eric
    Freytes, Donald O.
    Noggle, Scott
    Fossati, Valentina
    [J]. STEM CELL REPORTS, 2017, 8 (06): : 1516 - 1524
  • [38] Mechanotransductive Differentiation of Hair Follicle Stem Cells Derived from Aged Eyelid Skin into Corneal Endothelial-Like Cells
    Olszewski, Christian
    Maassen, Jessika
    Guenther, Rebecca
    Skazik-Voogt, Claudia
    Gutermuth, Angela
    [J]. STEM CELL REVIEWS AND REPORTS, 2022, 18 (05) : 1668 - 1685
  • [39] Directed differentiation of human pluripotent stem cells into epidermal stem and progenitor cells
    Sonya Ruiz-Torres
    Paul F. Lambert
    Kathryn A. Wikenheiser-Brokamp
    Susanne I. Wells
    [J]. Molecular Biology Reports, 2021, 48 : 6213 - 6222
  • [40] Descemet's Membrane Biomimetic Microtopography Differentiates Human Mesenchymal Stem Cells Into Corneal Endothelial-Like Cells
    Gutermuth, Angela
    Maassen, Jessika
    Harnisch, Emely
    Kuhlen, Daniel
    Sauer-Budge, Alexis
    Skazik-Voogt, Claudia
    Engelmann, Katrin
    [J]. CORNEA, 2019, 38 (01) : 110 - 119