Human iPS cells engender corneal epithelial stem cells with holoclone-forming capabilities

The future of eye reconstruction invariably includes stem cells transplantation. Corneal limbus, corneal stroma, trabeculum, retinal cells, optic nerve, and all structures that are irreversibly damaged and have no means to be repaired or replaced, through conventional treatment or surgery, represent targets for stem cell reconstruction. This review tries to answer the question if there is any clinical validation for stem therapies, so far, starting from the cornea and, on the path of light, arriving to the retina. The investigation covers the last 10 years of publications. From 2385 published sources, we found 56 clinical studies matching inclusion criteria, 39 involving cornea, and 17 involving retina. So far, corneal epithelial reconstruction seems well validated clinically. Enough clinical data are collected to allow some form of standardization for the stem cell transplant procedures. Cultivated limbal epithelial stem cells (CLET), simple limbal epithelial transplant (SLET), and oral mucosa transplantation are implemented worldwide. In comparison, far less patients are investigated in retinal stem reconstructions, with lower anatomical and clinical success, so far. Intravitreal, subretinal, and suprachoroidal approach for retinal stem therapies face specific challenges.

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Bone Morphogenetic Protein 4 (BMP4) Enhances the Differentiation of Human Induced Pluripotent Stem Cells into Limbal Progenitor Cells

Current Issues in Molecular Biology ◽

10.3390/cimb43030147 ◽

2021 ◽

Vol 43 (3) ◽

pp. 2124-2134

Author(s):

Hyun Soo Lee ◽

Jeewon Mok ◽

Choun-Ki Joo

Keyword(s):

Progenitor Cells ◽

Dermal Fibroblast ◽

Ips Cells ◽

Human Dermal Fibroblast ◽

Corneal Epithelial ◽

Specific Medium ◽

Morphogenetic Protein ◽

Human Ips Cells ◽

Limbal Stem Cell ◽

Induced Pluripotent

Corneal epithelium maintains visual acuity and is regenerated by the proliferation and differentiation of limbal progenitor cells. Transplantation of human limbal progenitor cells could restore the integrity and functionality of the corneal surface in patients with limbal stem cell deficiency. However, multiple protocols are employed to differentiate human induced pluripotent stem (iPS) cells into corneal epithelium or limbal progenitor cells. The aim of this study was to optimize a protocol that uses bone morphogenetic protein 4 (BMP4) and limbal cell-specific medium. Human dermal fibroblast-derived iPS cells were differentiated into limbal progenitor cells using limbal cell-specific (PI) medium and varying doses (1, 10, and 50 ng/mL) and durations (1, 3, and 10 days) of BMP4 treatment. Differentiated human iPS cells were analyzed by real-time polymerase chain reaction (RT-PCR), Western blotting, and immunocytochemical studies at 2 or 4 weeks after BMP4 treatment. Culturing human dermal fibroblast-derived iPS cells in limbal cell-specific medium and BMP4 gave rise to limbal progenitor and corneal epithelial-like cells. The optimal protocol of 10 ng/mL and three days of BMP4 treatment elicited significantly higher limbal progenitor marker (ABCG2, ∆Np63α) expression and less corneal epithelial cell marker (CK3, CK12) expression than the other combinations of BMP4 dose and duration. In conclusion, this study identified a successful reprogramming strategy to induce limbal progenitor cells from human iPS cells using limbal cell-specific medium and BMP4. Additionally, our experiments indicate that the optimal BMP4 dose and duration favor limbal progenitor cell differentiation over corneal epithelial cells and maintain the phenotype of limbal stem cells. These findings contribute to the development of therapies for limbal stem cell deficiency disorders.

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Isolated Corneal Epithelial Stem Cells Derived from Limbal Biopsies: Use of Lectin as a Marker for Identifying Transient Amplifying Cells

Stem Cells and Cancer Stem Cells,Volume 3 ◽

10.1007/978-94-007-2415-0_12 ◽

2011 ◽

pp. 125-138

Author(s):

Luciana Dini ◽

Cristian Vergallo

Keyword(s):

Stem Cells ◽

Epithelial Stem Cells ◽

Corneal Epithelial

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MicroRNA Profiling of Highly Enriched Human Corneal Epithelial Stem Cells by Small RNA Sequencing

Scientific Reports ◽

10.1038/s41598-020-64273-0 ◽

2020 ◽

Vol 10 (1) ◽

Cited By ~ 1

Author(s):

Lavanya Kalaimani ◽

Bharanidharan Devarajan ◽

Umadevi Subramanian ◽

Vanniarajan Ayyasamy ◽

Venkatesh Prajna Namperumalsamy ◽

...

Keyword(s):

Stem Cells ◽

Rna Sequencing ◽

Small Rna ◽

Small Rna Sequencing ◽

Epithelial Stem Cells ◽

Corneal Epithelial ◽

Microrna Profiling

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Corneal epithelial stem cells for corneal injury

Expert Opinion on Biological Therapy ◽

10.1080/14712598.2018.1508443 ◽

2018 ◽

Vol 18 (9) ◽

pp. 997-1003 ◽

Cited By ~ 5

Author(s):

Dominique Bremond-Gignac ◽

Henri Copin ◽

Moncef Benkhalifa

Keyword(s):

Stem Cells ◽

Epithelial Stem Cells ◽

Corneal Epithelial ◽

Corneal Injury

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Differentiation-related expression of a major 64K corneal keratin in vivo and in culture suggests limbal location of corneal epithelial stem cells.

The Journal of Cell Biology ◽

10.1083/jcb.103.1.49 ◽

1986 ◽

Vol 103 (1) ◽

pp. 49-62 ◽

Cited By ~ 830

Author(s):

A Schermer ◽

S Galvin ◽

T T Sun

Keyword(s):

Stem Cells ◽

Strong Support ◽

Transitional Zone ◽

Basal Cells ◽

Epithelial Stem Cells ◽

Corneal Epithelial ◽

Differentiated Cells ◽

Limbal Epithelium ◽

Cell Layers

In this paper we present keratin expression data that lend strong support to a model of corneal epithelial maturation in which the stem cells are located in the limbus, the transitional zone between cornea and conjunctiva. Using a new monoclonal antibody, AE5, which is highly specific for a 64,000-mol-wt corneal keratin, designated RK3, we demonstrate that this keratin is localized in all cell layers of rabbit corneal epithelium, but only in the suprabasal layers of the limbal epithelium. Analysis of cultured corneal keratinocytes showed that they express sequentially three major keratin pairs. Early cultures consisting of a monolayer of "basal" cells express mainly the 50/58K keratins, exponentially growing cells synthesize additional 48/56K keratins, and postconfluent, heavily stratified cultures begin to express the 55/64K corneal keratins. Cell separation experiments showed that basal cells isolated from postconfluent cultures contain predominantly the 50/58K pair, whereas suprabasal cells contain additional 55/64K and 48/56K pairs. Basal cells of the older, postconfluent cultures, however, can become AE5 positive, indicating that suprabasal location is not a prerequisite for the expression of the 64K keratin. Taken together, these results suggest that the acidic 55K and basic 64K keratins represent markers for an advanced stage of corneal epithelial differentiation. The fact that epithelial basal cells of central cornea but not those of the limbus possess the 64K keratin therefore indicates that corneal basal cells are in a more differentiated state than limbal basal cells. These findings, coupled with the known centripetal migration of corneal epithelial cells, strongly suggest that corneal epithelial stem cells are located in the limbus, and that corneal basal cells correspond to "transient amplifying cells" in the scheme of "stem cells----transient amplifying cells----terminally differentiated cells."

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Transcription Factor TCF4 Maintains the Properties of Human Corneal Epithelial Stem Cells

Stem Cells ◽

10.1002/stem.1032 ◽

2012 ◽

Vol 30 (4) ◽

pp. 753-761 ◽

Cited By ~ 39

Author(s):

Rong Lu ◽

Yangluowa Qu ◽

Jian Ge ◽

Lili Zhang ◽

Zhitao Su ◽

...

Keyword(s):

Stem Cells ◽

Transcription Factor ◽

Epithelial Stem Cells ◽

Corneal Epithelial

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Protection of corneal epithelial stem cells prevents ultraviolet A damage during corneal collagen cross-linking treatment for keratoconus

British Journal of Ophthalmology ◽

10.1136/bjophthalmol-2013-303816 ◽

2013 ◽

Vol 98 (2) ◽

pp. 270-274 ◽

Cited By ~ 12

Author(s):

Jonathan E Moore ◽

Sarah D Atkinson ◽

Dimitri T Azar ◽

Jenny Worthington ◽

C Stephen Downes ◽

...

Keyword(s):

Stem Cells ◽

Cross Linking ◽

Ultraviolet A ◽

Epithelial Stem Cells ◽

Corneal Epithelial ◽

Collagen Cross Linking ◽

Corneal Collagen

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Human induced pluripotent stem cells are capable of B-cell lymphopoiesis

Blood ◽

10.1182/blood-2010-08-299941 ◽

2011 ◽

Vol 117 (15) ◽

pp. 4008-4011 ◽

Cited By ~ 50

Author(s):

Lee Carpenter ◽

Ram Malladi ◽

Cheng-Tao Yang ◽

Anna French ◽

Katherine J. Pilkington ◽

...

Keyword(s):

Stem Cells ◽

B Cell ◽

Embryonic Stem ◽

Ips Cells ◽

Ips Cell ◽

Lymphoid Lineage ◽

Human Ips Cells ◽

Induced Pluripotent ◽

First Time ◽

Unique Potential

Abstract Induced pluripotent stem (iPS) cells offer a unique potential for understanding the molecular basis of disease and development. Here we have generated several human iPS cell lines, and we describe their pluripotent phenotype and ability to differentiate into erythroid cells, monocytes, and endothelial cells. More significantly, however, when these iPS cells were differentiated under conditions that promote lympho-hematopoiesis from human embryonic stem cells, we observed the formation of pre-B cells. These cells were CD45+CD19+CD10+ and were positive for transcripts Pax5, IL7αR, λ-like, and VpreB receptor. Although they were negative for surface IgM and CD5 expression, iPS-derived CD45+CD19+ cells also exhibited multiple genomic D-JH rearrangements, which supports a pre–B-cell identity. We therefore have been able to demonstrate, for the first time, that human iPS cells are able to undergo hematopoiesis that contributes to the B-cell lymphoid lineage.

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