Yan Zhang, Xiu-Li Hao, Shi-Fang Jia, Yan-Zhen Wen, Ying-Hui Wang
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Efficient genetic engineering of murine cochlear organoids
Organoid culture is a popular model to study gene function as the easy manipulating and time saving compared with in vivo experiments. This is widely used in auditory system for studying supporting cells (SCs) or hair cells (HCs) as only very few SCs or HCs can be harvested in both human and murine cochlea. However, the use of organoids is still a challenge due to the low efficiency in genetic modification. Here we took Lin28b as an example and compared Lin28b gain-of-function (GOF) and loss-of-function (LOF) with different genetic engineering methods and found that TetOn induced GOF or LOF was more efficient compared with lipofection or lentiviral transduction in the experimental conditions we used. Cell apoptosis in TetOn induction system was lowest compared with the other methods in this study. Our study is the first to compare the efficiency of different genetic engineering techniques in cochlear organoid culture, which may also apply to organoids established with other tissues.
期刊介绍:
Journal of Tissue Engineering and Regenerative Medicine publishes rapidly and rigorously peer-reviewed research papers, reviews, clinical case reports, perspectives, and short communications on topics relevant to the development of therapeutic approaches which combine stem or progenitor cells, biomaterials and scaffolds, growth factors and other bioactive agents, and their respective constructs. All papers should deal with research that has a direct or potential impact on the development of novel clinical approaches for the regeneration or repair of tissues and organs.
The journal is multidisciplinary, covering the combination of the principles of life sciences and engineering in efforts to advance medicine and clinical strategies. The journal focuses on the use of cells, materials, and biochemical/mechanical factors in the development of biological functional substitutes that restore, maintain, or improve tissue or organ function. The journal publishes research on any tissue or organ and covers all key aspects of the field, including the development of new biomaterials and processing of scaffolds; the use of different types of cells (mainly stem and progenitor cells) and their culture in specific bioreactors; studies in relevant animal models; and clinical trials in human patients performed under strict regulatory and ethical frameworks. Manuscripts describing the use of advanced methods for the characterization of engineered tissues are also of special interest to the journal readership.