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Induced pluripotent stem cells restore function in a human cell loss model of open-angle glaucoma.

Abu-Hassan DW, Li X, Ryan EI, Acott TS, Kelley MJ - Stem Cells (2015)

Bottom Line: A notable feature associated with glaucoma is outflow pathway cell loss.We then differentiated induced pluripotent stem cells (iPSCs) and used them to repopulate this cell depletion model.These differentiated cells (TM-like iPSCs) became similar to TM cells in both morphology and expression patterns.

View Article: PubMed Central - PubMed

Affiliation: Department of Ophthalmology, Casey Eye Institute, Oregon Health & Science University, Portland, Oregon, USA; Department of Biochemistry & Molecular Biology, Oregon Health & Science University, Portland, Oregon, USA; Department of Biochemistry & Physiology, University of Jordan, Amman, Jordan.

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Replacement of saponin-depleted cells with human cultured TM cells. (A): Detailed schematic of treatment protocol. (B): Frontal section showing penetration of QDot (red) labeled HTM cells to all levels of the TM after transplantation. Scale bar is 100 µm. (C): TM beams showing blue autofluorescence from collagen and elastic fibers (TM Beams), CD44 immunohistochemistry (green) to label cell surfaces, and QDot (red) labeled transplanted TM cells. White dashes enclose individual cells, which contain QDots indicating that they were transplanted. Dashes were drawn based on Z-stack three-dimensional scans to identify individual QDot-labeled cells. The scale bar is 100 µm. (D): Transplanted replacement HTM cells (added at the time indicated) restored the intraocular pressure homeostatic response to 2× pressure elevation, which had been compromised by saponin treatment. Line shows mean for six experiments using separate anterior segments and error bars represent the SEM with significance by one-way ANOVA where *, p < .05 and **, p < .001. Abbreviations: HTM, human trabecular meshwork; JCT, juxtacanalicular region; SC, Schlemm's canal; TM, trabecular meshwork.
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fig02: Replacement of saponin-depleted cells with human cultured TM cells. (A): Detailed schematic of treatment protocol. (B): Frontal section showing penetration of QDot (red) labeled HTM cells to all levels of the TM after transplantation. Scale bar is 100 µm. (C): TM beams showing blue autofluorescence from collagen and elastic fibers (TM Beams), CD44 immunohistochemistry (green) to label cell surfaces, and QDot (red) labeled transplanted TM cells. White dashes enclose individual cells, which contain QDots indicating that they were transplanted. Dashes were drawn based on Z-stack three-dimensional scans to identify individual QDot-labeled cells. The scale bar is 100 µm. (D): Transplanted replacement HTM cells (added at the time indicated) restored the intraocular pressure homeostatic response to 2× pressure elevation, which had been compromised by saponin treatment. Line shows mean for six experiments using separate anterior segments and error bars represent the SEM with significance by one-way ANOVA where *, p < .05 and **, p < .001. Abbreviations: HTM, human trabecular meshwork; JCT, juxtacanalicular region; SC, Schlemm's canal; TM, trabecular meshwork.

Mentions: Although anterior segments exposed to saponin lost the ability to adjust the outflow resistance when subjected to a 2× pressure challenge, this capability was regained when cultured human TM cells were added back and allowed to attach and integrate into the anterior segments (Fig. 2A–2C). After saponin-treatment and assessment of response to a 2× pressure challenge, human TM cells that had been labeled with red fluorescent QDots for identification purposes were injected into the perfusion line, allowed to flow into the TM at 0.5× pressure for 2 hours, and then the pressure was reduced to 0 mmHg overnight to facilitate cell attachment. Perfusion pressure was then reinitiated, maintained at 1× to re-establish the baseline flow, then increased to 2×, and the flow was measured for several days to assess capability to exhibit an IOP homeostatic response (Fig. 2D). When examined by confocal microscopy (Fig. 2B, 2C), QDot-labeled transplanted TM cells were observed throughout the TM. The transplanted cells appeared to have attached to the TM beams and to inner layers of TM including the JCT (Fig. 2B, 2C). Although the IOP homeostatic response to 2× pressure was lost after saponin treatment, it was regained after the TM cells were added back and allowed to integrate into the open areas (Fig. 2D).


Induced pluripotent stem cells restore function in a human cell loss model of open-angle glaucoma.

Abu-Hassan DW, Li X, Ryan EI, Acott TS, Kelley MJ - Stem Cells (2015)

Replacement of saponin-depleted cells with human cultured TM cells. (A): Detailed schematic of treatment protocol. (B): Frontal section showing penetration of QDot (red) labeled HTM cells to all levels of the TM after transplantation. Scale bar is 100 µm. (C): TM beams showing blue autofluorescence from collagen and elastic fibers (TM Beams), CD44 immunohistochemistry (green) to label cell surfaces, and QDot (red) labeled transplanted TM cells. White dashes enclose individual cells, which contain QDots indicating that they were transplanted. Dashes were drawn based on Z-stack three-dimensional scans to identify individual QDot-labeled cells. The scale bar is 100 µm. (D): Transplanted replacement HTM cells (added at the time indicated) restored the intraocular pressure homeostatic response to 2× pressure elevation, which had been compromised by saponin treatment. Line shows mean for six experiments using separate anterior segments and error bars represent the SEM with significance by one-way ANOVA where *, p < .05 and **, p < .001. Abbreviations: HTM, human trabecular meshwork; JCT, juxtacanalicular region; SC, Schlemm's canal; TM, trabecular meshwork.
© Copyright Policy - open-access
Related In: Results  -  Collection

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fig02: Replacement of saponin-depleted cells with human cultured TM cells. (A): Detailed schematic of treatment protocol. (B): Frontal section showing penetration of QDot (red) labeled HTM cells to all levels of the TM after transplantation. Scale bar is 100 µm. (C): TM beams showing blue autofluorescence from collagen and elastic fibers (TM Beams), CD44 immunohistochemistry (green) to label cell surfaces, and QDot (red) labeled transplanted TM cells. White dashes enclose individual cells, which contain QDots indicating that they were transplanted. Dashes were drawn based on Z-stack three-dimensional scans to identify individual QDot-labeled cells. The scale bar is 100 µm. (D): Transplanted replacement HTM cells (added at the time indicated) restored the intraocular pressure homeostatic response to 2× pressure elevation, which had been compromised by saponin treatment. Line shows mean for six experiments using separate anterior segments and error bars represent the SEM with significance by one-way ANOVA where *, p < .05 and **, p < .001. Abbreviations: HTM, human trabecular meshwork; JCT, juxtacanalicular region; SC, Schlemm's canal; TM, trabecular meshwork.
Mentions: Although anterior segments exposed to saponin lost the ability to adjust the outflow resistance when subjected to a 2× pressure challenge, this capability was regained when cultured human TM cells were added back and allowed to attach and integrate into the anterior segments (Fig. 2A–2C). After saponin-treatment and assessment of response to a 2× pressure challenge, human TM cells that had been labeled with red fluorescent QDots for identification purposes were injected into the perfusion line, allowed to flow into the TM at 0.5× pressure for 2 hours, and then the pressure was reduced to 0 mmHg overnight to facilitate cell attachment. Perfusion pressure was then reinitiated, maintained at 1× to re-establish the baseline flow, then increased to 2×, and the flow was measured for several days to assess capability to exhibit an IOP homeostatic response (Fig. 2D). When examined by confocal microscopy (Fig. 2B, 2C), QDot-labeled transplanted TM cells were observed throughout the TM. The transplanted cells appeared to have attached to the TM beams and to inner layers of TM including the JCT (Fig. 2B, 2C). Although the IOP homeostatic response to 2× pressure was lost after saponin treatment, it was regained after the TM cells were added back and allowed to integrate into the open areas (Fig. 2D).

Bottom Line: A notable feature associated with glaucoma is outflow pathway cell loss.We then differentiated induced pluripotent stem cells (iPSCs) and used them to repopulate this cell depletion model.These differentiated cells (TM-like iPSCs) became similar to TM cells in both morphology and expression patterns.

View Article: PubMed Central - PubMed

Affiliation: Department of Ophthalmology, Casey Eye Institute, Oregon Health & Science University, Portland, Oregon, USA; Department of Biochemistry & Molecular Biology, Oregon Health & Science University, Portland, Oregon, USA; Department of Biochemistry & Physiology, University of Jordan, Amman, Jordan.

Show MeSH
Related in: MedlinePlus