Peggy Arthur1, Sangeetha Kandoi2,3, Li Sun4,5, Anil Kalvala1, Shallu Kutlehria1, Santanu Bhattacharya6,7, Tanmay Kulkarni6, Ramesh Nimma1, Yan Li8, Deepak A Lamba9, Mandip Singh10. 1. College of Pharmacy and Pharmacological Sciences, Florida A&M University, Tallahassee, FL, USA. 2. Department of Ophthalmology, University of California San Francisco, San Francisco, CA, USA. 3. Eli and Edythe Broad Center of Regeneration Medicine and Stem Cell Research, University of California San Francisco, San Francisco, CA, USA. 4. Department of Chemical and Biomedical Engineering, FAMU-FSU College of Engineering, Florida State University, Tallahassee, FL, USA. 5. Department of Biomedical Sciences, College of Medicine, Florida State University, Tallahassee, FL, USA. 6. Department of Biochemistry and Molecular Biology, Mayo College of Medicine and Science, Jacksonville, FL, USA. 7. Department of Physiology and Biomedical Engineering, Mayo College of Medicine and Science, Jacksonville, FL, USA. 8. Department of Chemical and Biomedical Engineering, FAMU-FSU College of Engineering, Florida State University, Tallahassee, FL, USA. yli@eng.famu.fsu.edu. 9. Department of Ophthalmology, University of California San Francisco, San Francisco, CA, USA. deepak.lamba@ucsf.edu. 10. College of Pharmacy and Pharmacological Sciences, Florida A&M University, Tallahassee, FL, USA. mandip.sachdeva@famu.edu.
Abstract
PURPOSE: There is a growing interest in extracellular vesicles (EVs) for ocular applications as therapeutics, biomarkers, and drug delivery vehicles. EVs secreted from mesenchymal stem cells (MSCs) have shown to provide therapeutic benefits in ocular conditions. However, very little is known about the properties of bioreactor cultured-3D human retinal organoids secreted EVs. This study provides a comprehensive morphological, nanomechanical, molecular, and proteomic characterization of retinal organoid EVs and compares it with human umbilical cord (hUC) MSCs. METHODS: The morphology and nanomechanical properties of retinal organoid EVs were assessed using Nanoparticle tracking analysis (NTA) and Atomic force microscopy (AFM). Gene expression analysis of exosome biogenesis of early and late retinal organoids were compared using qPCR. The protein profile of the EVs were analyzed with proteomic tools. RESULTS: NTA indicated the average size of EV as 100-250 nm. A high expression of exosome biogenesis genes was observed in late retinal organoids EVs. Immunoblot analysis showed highly expressed exosomal markers in late retinal organoids EVs compared to early retinal organoids EVs. Protein profiling of retinal organoid EVs displayed a higher differential expression of retinal function-related proteins and EV biogenesis proteins than hUCMSC EVs, implicating that the use of retinal organoid EVs may have a superior therapeutic effect on retinal disorders. CONCLUSION: This study provides supplementary knowledge on the properties of retinal organoid EVs and suggests their potential use in the diagnostic and therapeutic treatments for ocular diseases.
PURPOSE: There is a growing interest in extracellular vesicles (EVs) for ocular applications as therapeutics, biomarkers, and drug delivery vehicles. EVs secreted from mesenchymal stem cells (MSCs) have shown to provide therapeutic benefits in ocular conditions. However, very little is known about the properties of bioreactor cultured-3D human retinal organoids secreted EVs. This study provides a comprehensive morphological, nanomechanical, molecular, and proteomic characterization of retinal organoid EVs and compares it with human umbilical cord (hUC) MSCs. METHODS: The morphology and nanomechanical properties of retinal organoid EVs were assessed using Nanoparticle tracking analysis (NTA) and Atomic force microscopy (AFM). Gene expression analysis of exosome biogenesis of early and late retinal organoids were compared using qPCR. The protein profile of the EVs were analyzed with proteomic tools. RESULTS: NTA indicated the average size of EV as 100-250 nm. A high expression of exosome biogenesis genes was observed in late retinal organoids EVs. Immunoblot analysis showed highly expressed exosomal markers in late retinal organoids EVs compared to early retinal organoids EVs. Protein profiling of retinal organoid EVs displayed a higher differential expression of retinal function-related proteins and EV biogenesis proteins than hUCMSC EVs, implicating that the use of retinal organoid EVs may have a superior therapeutic effect on retinal disorders. CONCLUSION: This study provides supplementary knowledge on the properties of retinal organoid EVs and suggests their potential use in the diagnostic and therapeutic treatments for ocular diseases.
Authors: David A Parfitt; Amelia Lane; Conor M Ramsden; Amanda-Jayne F Carr; Peter M Munro; Katarina Jovanovic; Nele Schwarz; Naheed Kanuga; Manickam N Muthiah; Sarah Hull; Jean-Marc Gallo; Lyndon da Cruz; Anthony T Moore; Alison J Hardcastle; Peter J Coffey; Michael E Cheetham Journal: Cell Stem Cell Date: 2016-04-14 Impact factor: 24.633
Authors: Kiara C Eldred; Sarah E Hadyniak; Katarzyna A Hussey; Boris Brenerman; Ping-Wu Zhang; Xitiz Chamling; Valentin M Sluch; Derek S Welsbie; Samer Hattar; James Taylor; Karl Wahlin; Donald J Zack; Robert J Johnston Journal: Science Date: 2018-10-12 Impact factor: 47.728
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