Human induced pluripotent stem cell line HMUi001-A derived from corneal stromal cells

Bikkuzin, Timur; Shi, Yan; Sun, Baoqi; Guo, Yiyuan; Jin, Xin; Han, Zhenbo; Pavlov, V. N.; Zhang, Hong

doi:10.1016/j.scr.2019.101409

Cited by 4 publications

(3 citation statements)

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“…To date, a variety of cell types have been reported to be suitable for reprogramming, including blood cells (Chen et al, 2019), adult stem cells (Jiang et al, 2019) and even cancer cells (Bang et al, 2019). With respect to corneal stromal cells, most previous studies used reprogrammed corneal fibroblasts to generate iPSCs (Greene et al, 2014;Bikkuzin et al, 2019). Chien et al (2012) reported that using the four-gene delivery method (Oct4/Sox2/Klf4/c-Myc), they successfully created iPSCs by dedifferentiating corneal keratocytes isolated from corneal tissues.…”

Section: Discussionmentioning

confidence: 99%

“…When keratocytes are removed from stromal tissue and cultured in serum-containing medium, they exhibit the morphological characteristics of fibroblasts or myofibroblasts and express fibronectin and α-SMA (Kumar et al, 2016). It is well known that corneal fibroblasts can be successfully reprogrammed to form induced pluripotent stem cells (iPSCs) by transfecting with transcription factor genes (OSKM: Oct4, Sox2, Klf4, c-Myc) (Bikkuzin et al, 2019). However, the reprogramming of corneal keratocytes has not yet been investigated.…”

Section: Introductionmentioning

confidence: 99%

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Mechanotransduction Regulates Reprogramming Enhancement in Adherent 3D Keratocyte Cultures

Ding²,

Guo³

et al. 2021

Front. Bioeng. Biotechnol.

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Suspended spheroid culture using ultralow attachment plates (ULAPs) is reported to effect corneal fibroblast reprogramming. Polydimethylsiloxane (PDMS), with hydrophobic and soft substrate properties, facilitates adherent spheroid formation that promotes cellular physical reprogramming into stem-like cells without using transcription factors. However, it is still unknown whether the biophysical properties of PDMS have the same effect on adult human corneal keratocyte reprogramming. Here, PDMS and essential 8 (E8) medium were utilized to culture keratocyte spheroids and fibroblast spheroids, and the reprogramming results were compared. We provide insights into the probable mechanisms of the PDMS effect on spheroids. qPCR analysis showed that the expression of some stem cell marker genes (OCT4, NANOG, SOX2, KLF4, CMYC, ABCG2 and PAX6) was significantly greater in keratocyte spheroids than in fibroblast spheroids. The endogenous level of stemness transcription factors (OCT4, NANOG, SOX2, KLF4 and CMYC) was higher in keratocytes than in fibroblasts. Immunofluorescence staining revealed Klf4, Nanog, Sox2, ABCG2 and Pax6 were positively stained in adherent 3D spheroids but weakly or negatively stained in adherent 2D cells. Furthermore, OCT4, NANOG, SOX2, KLF4, HNK1, ABCG2 and PAX6 gene expression was significantly higher in adherent 3D spheroids than in adherent 2D cells. Meanwhile, SOX2, ABCG2 and PAX6 were more upregulated in adherent 3D spheroids than in suspended 3D spheroids. The RNA-seq analysis suggested that regulation of the actin cytoskeleton, TGFβ/BMP and HIF-1 signaling pathways induced changes in mechanotransduction, the mesenchymal-to-epithelial transition and hypoxia, which might be responsible for the effect of PDMS on facilitating reprogramming. In conclusion, compared to corneal fibroblasts, keratocytes were more susceptible to reprogramming due to higher levels of endogenous stemness transcription factors. Spheroid culture of keratocytes using PDMS had a positive impact on promoting the expression of some stem cell markers. PDMS, as a substrate to form spheroids, was better able to promote reprogramming than ULAPs. These results indicated that the physiological cells and culture conditions herein enhance reprogramming. Therefore, adherent spheroid culture of keratocytes using PDMS is a promising strategy to more safely promote reprogramming, suggesting its potential application for developing clinical implants in tissue engineering and regenerative medicine.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Mechanotransduction Regulates Reprogramming Enhancement in Adherent 3D Keratocyte Cultures

Ding²,

Guo³

et al. 2021

Front. Bioeng. Biotechnol.

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“…Due to their numerous advantages, iPSCs are widely regarded as an optimal source of stem cells. The establishment of the human corneal stromal iPSCs line (HMUi001-A) has been achieved [49], but further validation of its functionality is required. In general, the transplantation treatment using CSSCs is still in the stage of animal experimentation and preclinical research, with a long way to go before human clinical trials can be conducted.…”

Section: Stem Cell Therapy For Corneal Diseasesmentioning

confidence: 99%

The next‐generation therapies in ophthalmology for blindness worldwide

Xu,

Zhang,

Jiang

et al. 2024

Eye & ENT Research

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With the rapid and groundbreaking development in 21st century medicine on a global scale, new possibilities have emerged for addressing eye diseases, that can be blindness, and that conventional pharmaceuticals and surgical interventions have not hitherto been able to adequately treat. Gene enhancement/supplementation, gene editing, and stem cell therapies have now emerged as key subdisciplines. Here we discuss the current state and prospects of regenerative therapy in the field of ophthalmology, with a primary focus on diseases affecting the cornea, retina, and optic nerve. Our review summarizes the latest advances, challenges, and opportunities in these fields, as well as the potential applications and limitations of different strategies. The review also highlights the importance of interdisciplinary and collaborative innovation models for achieving breakthroughs in therapeutic development for sight‐loss diseases worldwide.

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