BAM15 attenuates transportation-induced apoptosis in iPS-differentiated retinal tissue

Abstract

Background: BAM15 is a novel mitochondrial protonophore uncoupler capable of protecting mammals from acute renal ischemic-reperfusion injury and cold-induced microtubule damage. The purpose of our study was to investigate the effect of BAM15 on apoptosis during 5-day transportation of human-induced pluripotent stem (hiPS)-differentiated retinal tissue. Methods: Retinal tissues of 30 days and 60 days were transported with or without BAM15 for 5 days in the laboratory or by real express. Immunofluorescence staining of apoptosis marker cleaved caspase3, proliferation marker Ki67, and neural axon marker NEFL was performed. And expression of apoptotic-related factors p53, NFkappaB, and TNF-a was detected by real-time PCR. Also, location of ganglion cells, photoreceptor cells, amacrine cells, and precursors of neuronal cell types in retinal tissue was stained by immunofluorescence after transportation. Furthermore, cell viability was assessed by CCK8 assay.Results: Results showed transportation remarkably intensified expression of apoptotic factor cleaved caspase3, p53, NFkappaB, and TNF-a, which could be reduced by supplement of BAM15. In addition, neurons were severely injured after transportation, with axons manifesting disrupted and tortuous by staining NEFL. And the addition of BAM15 in transportation was able to protect neuronal structure and increase cell viability without affecting subtypes cells location of retinal tissue. Conclusions: BAM15 might be used as a protective reagent on apoptosis during transporting retinal tissues, holding great potential in research and clinical applications.

BAM15, a Mitochondrial Uncoupling Agent, Attenuates Inflammation in the LPS Injection Mouse Model: An Adjunctive Anti-Inflammation on Macrophages and Hepatocytes

“…Not only energy generation and thermogenesis but also an impact of mitochondria toward immune responses is well described [15]. Here, a mitochondrial uncoupling agent (BAM15) attenuated inflammation via the suppression of cell energy status on mitochondria and glycolysis activity, supported by previous publications [23, 60]. Our model demonstrated that BAM15 increased cell energy status at the early time point (<1 h after BAM15) similar to the previous reports [29, 30]; however, the cell energy was reduced at 24 h after BAM15 resulting in the anti-inflammatory characteristics.…”

BAM15 treats mouse sepsis and sepsis-AKI, linking circulating mitochondrial DNA and tubule reactive oxygen species

“…Interestingly, BAM15 did not have much effect on other organ injury nor did it alter TNFα or IL-17 levels. The lack of effect on TNFα is puzzling, as a previous report found that BAM15 reduced increased expression of TNF-α in retinal tissues during donor organ transportation (43).…”

Comparison of DNA stability and its related genes of neurons derived from induced pluripotent stem cells and primary retinal neurons

“…The ESCs (H9 line) were differentiated to retinal tissue in accordance with previous established protocol (Tang et al, 2019; Zhong et al, 2014). Briefly, on Day 0 of differentiation, the cells were dissociated into small clumps and cultured in a low‐touch dish with mTeSR™ 1 medium [85850; STEMCELL] and supplemented with 10 μM Blebbistatin [B0560; Sigma‐Aldrich].…”

“…The ESCs (H9 line) were differentiated to retinal tissue in accordance with previous established protocol (Tang et al, 2019;Zhong et al, 2014). and medium was changed every 2 days.…”