Effects of Cord Blood Serum (CBS) on viability of retinal Müller glial cells under in vitro injury

Abstract

Oxidative stress and inflammation determine retinal ganglion cell degeneration, leading to retinal impairment and vision loss. Mü ller glial cells regulate retinal repair under injury, through gliosis. Meanwhile, reactive gliosis can turn in pathological effects, contributing to neurodegeneration. In the present study, we tested whether Cord Blood Serum (CBS), rich of growth factors, might improve the viability of Mü ller cells under in vitro damage. BDNF, NGF, TGF-α, GDNF and EGF levels were measured in CBS samples by Human Magnetic Luminex Assay. CBS effects were evaluated on rat (rMC-1) and human (MIO-M1) Mü ller cells, under H 2 O 2 and IL-1β damage. Cells grown with FBS or CBS both at 5% were exposed to stress and analyzed in terms of cell viability, GFAP, IL-6 and TNF-α expression. CBS was also administrated after treatment with K252a, inhibitor of the neurotrophin receptor Trk. Cell viability of rMC-1 and MIO-M1 resulted significantly improved when pretreated with CBS and exposed to H 2 O 2 and IL-1β, in comparison to the standard culture with FBS. Accordingly, the gliosis marker GFAP resulted down-regulated following CBS priming. In parallel, we observed a lower expression of the inflammatory mediators in rMC-1 (TNF-α) and MIO-M1 (IL-6, TNFα), especially in presence of inflammatory damage. Trk inhibition through K252a administration impaired the effects of CBS under stress conditions on MIO-M1 and rMC-1 viability, not significantly different from FBS condition. CBS is enriched with neurotrophins and its administration to rMC-1 and MIO-M1 attenuates the cytotoxic effects of H 2 O 2 and IL-1β. Moreover, the decrease of the main markers of gliosis and inflammation suggests a promising use of CBS for neuroprotection aims. This study is a preliminary basis that prompts future investigations to deeply explore and confirm the CBS potential.

Human glial müller and umbilical vein endothelial cell coculture as an in vitro model to investigate retinal oxidative damage. A morphological and molecular assessment

“…We next investigated the gene expression of CD31 and GFAP in MIO‐M1/HUVEC coculture at p4 and p8 under oxidative stress conditions. Based on cell viability data and on our previous findings (Ciavarella et al, 2020), H 2 O 2 was used at 500 μM for 24 h. We observed up‐regulation of CD31 mRNA in MIO‐M1/HUVEC coculture exposed to H 2 O 2 in comparison to untreated controls (30‐fold increase in MIO‐M1/HUVEC p4, p 0.076; 100‐fold increase in MIO‐M1/HUVEC p8, p = 0.033, unpaired t test) (Figure 8a), suggesting a potential modulation of cell damage through the stimulation of the endothelial differentiation and neo‐vascularization. However, the low expression of GFAP transcript indicated that this response was not accompanied by MIO‐M1 activation and cell suffering (Figure 8b).…”

“…We next investigated the gene expression of CD31 and GFAP in MIO-M1/HUVEC coculture at p4 and p8 under oxidative stress conditions. Based on cell viability data and on our previous findings(Ciavarella et al, 2020), H 2 O 2 was used at 500 μM for 24 h. We observed up-regulation of CD31 mRNA in MIO-M1/HUVEC coculture exposed to H 2 O 2 in comparison to untreated controls (30-fold increase in MIO-M1/HUVEC p4, p 0.076; 100-fold increase in MIO-M1/HUVEC p8, p = 0.033, unpaired t test)…”

Impact of Freeze-Drying on Cord Blood (CB), Serum (S), and Platelet-Rich Plasma (CB-PRP) Preparations on Growth Factor Content and In Vitro Cell Wound Healing

“…Previous research showed the efficacy of cord blood serum (CB-S) at ameliorating the effects of oxidative stress and inflammation on the viability of rat and human Muller cells [27] by modulating the state of activation which typically occurs when glial cells are exposed to tissue damage. In the present study, the effect of lyophilized products from CB as a source was explored in MIO-M1 cells in terms of wound healing.…”

Harnessing Astrocytes and Müller Glial Cells in the Retina for Survival and Regeneration of Retinal Ganglion Cells

“…It is also critical to understand whether EGFR signaling can induce Müller cells to become like A2 astrocytes, as they undergo reactive gliosis along with retinal astrocytes in response to injury. Additionally, there has been increasing attention to the possibility to deliver neurotrophic factors by eye drops, and clinical studies suggest that this could represent a safe and effective strategy for treating retinal degenerative diseases [42,[90][91][92][93][94]. Hence, future research should evaluate the efficacy of EGFR ligand-based eye drops after ONC and investigate whether the topical delivery of EGFR ligands could generate growthsupportive phenotypes of retinal astrocytes and Müller cells.…”