September 2016
Volume 57, Issue 12
Open Access
ARVO Annual Meeting Abstract  |   September 2016
iPSC-derived TM cells promote proliferation of primary TM cells
Author Affiliations & Notes
  • Wei Zhu
    Department of Ophthalmology, University of Iowa, Iowa, Iowa, United States
    Iowa city veterans affairs medical center, Center for the prevention and treatment of visual loss, Iowa City, Iowa, United States
  • Oliver W Gramlich
    Department of Ophthalmology, University of Iowa, Iowa, Iowa, United States
    Iowa city veterans affairs medical center, Center for the prevention and treatment of visual loss, Iowa City, Iowa, United States
  • Budd A Tucker
    Department of Ophthalmology, University of Iowa, Iowa, Iowa, United States
  • Markus H Kuehn
    Department of Ophthalmology, University of Iowa, Iowa, Iowa, United States
    Iowa city veterans affairs medical center, Center for the prevention and treatment of visual loss, Iowa City, Iowa, United States
  • Footnotes
    Commercial Relationships   Wei Zhu, None; Oliver Gramlich, None; Budd Tucker, None; Markus Kuehn, None
  • Footnotes
    Support  None
Investigative Ophthalmology & Visual Science September 2016, Vol.57, 4680. doi:
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    • Get Citation

      Wei Zhu, Oliver W Gramlich, Budd A Tucker, Markus H Kuehn; iPSC-derived TM cells promote proliferation of primary TM cells. Invest. Ophthalmol. Vis. Sci. 2016;57(12):4680.

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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract

Purpose : We previously demonstrated that transplantation of induced pluripotent stem cell-derived trabecular meshwork cells (iPSC-TM) restores normal trabecular meshwork (TM) function in MYOCY437H transgenic mice. The goal of this study was to investigate the interactions between iPSC-TM and primary TM cells (pTM) in vitro and using perfusion cultured human eyes.

Methods : 50,000 mouse pTM cells were transfected with Ad5RSVmyocilinY437HHisFlag and subsequently either co-cultured with 50,000 purified mouse dsRed+ iPSC-TM either allowing cell-cell contact, separated using transwell inserts, or grown in cell culture media pre-conditioned by iPSC-TM. After 4 days the surviving dsRed-negative cells were counted by flow cytometry. The proliferation rate was analyzed based upon BrdU incorporation within a 2-hour period. Additionally 5 pairs of human donor eyes (83±9.6 yrs old), maintained in a perfusion organ culture system, received transplants of 200,000 human dsRed-positive iPSC-TM. After 2 weeks in culture, TM cells were isolated and counted by flow cytometry and the tissue was evaluated using histochemical and immunohistochemical methods.

Results : After 4 days in culture the number of mouse pTM cells maintained alone increased by 38.7%±6.0 whereas direct co-culture of pTM and iPSC-TM resulted in a 234.4%±68 increase (P=0.0039). The fraction of BrdU positive pTM cells in co-cultures was significant higher than that measured in controls (32.9% vs 21.6%, P=0.03). Significantly heightened proliferation rates were not observed in co-culture experiments when iPSC-TM were maintained in cell culture inserts (19.7%, P=0.16) or in iPSC-TM conditioned media (25.3%, P=0.08). Similar findings were also obtained in human eyes using a perfusion organ culture system. 4 out of 5 donors responded with vigorous TM cell proliferation upon transplantation of iPSC-TM. Although only a small number of surviving iPSC-TM was detected these eyes displayed a 2.1±0.98 fold increase in TM cellularity when compared to the untreated contralateral eye (P=0.035).

Conclusions : These studies demonstrate that iPSC-TM induce a proliferative response in both human and mouse TM cells that requires direct cell-cell contact. Importantly, TM cell proliferation can also be induced in aged human donor eyes, suggesting that this is a general response and not specifically related to myocilin mutation induced TM abnormalities.

This is an abstract that was submitted for the 2016 ARVO Annual Meeting, held in Seattle, Wash., May 1-5, 2016.

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