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    Neural Engineering

    (L-450) The joint contributionsof Müller Glia and astrocytes to the inner blood retinal barrier

    Friday, October 13, 2023
    3:30 PM – 4:30 PM PDT
    Location: Exhibit Hall - Row L - Poster # 450
    Introduction::

    Diabetic retinopathy is a leading eye disorder that leads to vision loss and blindness in many American diabetic adults. It is also the leading cause of new cases of blindness for people aged 20 to 74 years. Chronically elevated levels of glucose alter the integrity of the inner blood retinal barrier (iBRB), a neurovascular tissue composed of endothelial cells (EC), pericytes, astrocytes (AC), and Müller glia (MG)1 (Figure 1). This project examined the contributions of AC and MG to iBRB integrity by comparing cell morphology in a tri- and co-culture system with EC as well as permeability and resistivity.  



    Materials and Methods::  Cultured Primary Muller Glia, ndothelial Cell Culture (ThermoFisher M200500), Primary Adult rat retinal astrocytes (RA-r) (ScienCell, R1870) calculated Cell Area and Cell Shape Index (CSI) using ImageJ. We then culture Endothelial Cell-type culture media, and Liposaccharide (LPS) was used as a positive control. Measured growth over a span of three days. Also, measured Transepithelial/transendothelial electrical resistance (TEER) relationship between MG and astrocytes using EVOM device. 

    Results, Conclusions, and Discussions::

    Results: MG area in the presence of EC was significantly larger than AC while AC morphology was largely unchanged when cultured with EC in comparison to MG. At 24 hours, we see there is significance in the area for MG among control, EC and LPS. For the astrocytes, there was no significance in the area except for hour 6 (Figure 2). Astrocytes did however show a significance in their CSI in relation to MG. This highlights that there was a change in morphology in the ACs in the early hours. This data illustrates that ACs and MGs both have different individual contributions to the iBRB. The MG and EC combo conditions also show a significant relationship between the cells TEER and how they play a role in each other’s stability.


     


    Conclusions: Our results show a significant relationship between MG and EC that remains understudied compared to AC and EC.  Future tests will examine the relative contribution of MG and AC to iBRB integrity to help develop more appropriate disease models for diabetic retinopathy that aid development of therapies.  



    Conclusions:     Our results show a significant relationship between MG and EC that remains understudied compared to AC and EC.      Future tests will examine the relative contribution of MG and AC to iBRB integrity to help develop more appropriate disease models for diabetic retinopathy that aid development of therapies.  



    Acknowledgements (Optional): : Juan Pena and Dr. Maribel Vazquez

    References (Optional): :

    1 Pena, J.S.; Vazquez, M., 'Harnessing the Neuroprotective Behaviors of Müller Glia for Retinal Repair,' Frontiers in Bioscience-Landmark 2022 27(6), 169-181. https://www.imrpress.com/journal/FBL/27/6/10.31083/j.fbl2706169


    2 Castro, N.; Cohen, R.; Vazquez, M., 'Re: "Organ-On-A-Chip Technologies for Advanced Blood-Retinal Barrier Models," by Ragelle et al,' Journal of Ocular Pharmacology and Therapeutics 2022 Jun;38(5):329-330. https://pubmed.ncbi.nlm.nih.gov/35687506/