Abstract
BackgroundBAM15 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.MethodsRetinal 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.ResultsResults 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.ConclusionsBAM15 might be used as a protective reagent on apoptosis during transporting retinal tissues, holding great potential in research and clinical applications.
Highlights
BAM15 is a novel mitochondrial protonophore uncoupler capable of protecting mammals from acute renal ischemic-reperfusion injury and cold-induced microtubule damage
State Key Laboratory of Ophthalmology, Guangdong Provincial Key Laboratory of Ophthalmology and Visual Science, Zhongshan Ophthalmic Center, Sun Yat-sen University, Guangzhou, Guangdong 510060, China organs or stem cells [5, 7,8,9,10], we focused on seeking appropriate anti-apoptotic medium for the first time on transporting human-induced pluripotent stem cell (hiPSC)-differentiated retinal tissues
Influence of transportation duration and temperature on iPS-induced 3-D retinal tissues The experimental procedures are as follows (Fig. 1a): hiPSC was differentiated into three-dimensional retinal tissues, transported by mimic transportation or by real express transportation, and evaluated after recovery for 5 days. hiPSC was differentiated into horseshoe-shaped neural retina (NR) structures (Fig. 1b), which was isolated on D28 for long-term suspension culture
Summary
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. It has been a common practice to transport tissues among sites of collection, processing, and clinical areas for application [4]. It is reported stem cells undergo apoptosis during transportation [5]. Important considerations related to transportation include density of tissues, shipment duration, temperature management, and transportation medium [6]. Since the former three considerations have been investigated previously on other. Different from the traditional uncouplers, Tang et al Stem Cell Research & Therapy (2019) 10:64
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