Petrović Antonela, Ban Jelena, Tomljanović Ivana, Pongrac Marta, Ivaničić Matea, Mikašinović Sanja, Mladinic Miranda
Laboratory for Molecular Neurobiology, Department of Biotechnology, University of Rijeka, Rijeka, Croatia.
Front Cell Neurosci. 2021 Mar 18;15:661492. doi: 10.3389/fncel.2021.661492. eCollection 2021.
Primary dissociated neuronal cultures have become a standard model for studying central nervous system (CNS) development. Such cultures are predominantly prepared from the hippocampus or cortex of rodents (mice and rats), while other mammals are less used. Here, we describe the establishment and extensive characterization of the primary dissociated neuronal cultures derived from the cortex of the gray South American short-tailed opossums, . Opossums are unique in their ability to fully regenerate their CNS after an injury during their early postnatal development. Thus, we used cortex of postnatal day (P) 3-5 opossum to establish long-surviving and nearly pure neuronal cultures, as well as mixed cultures composed of radial glia cells (RGCs) in which their neurogenic and gliogenic potential was confirmed. Both types of cultures can survive for more than 1 month . We also prepared neuronal cultures from the P16-18 opossum cortex, which were composed of astrocytes and microglia, in addition to neurons. The long-surviving opossum primary dissociated neuronal cultures represent a novel mammalian platform particularly useful to study CNS development and regeneration.
原代解离神经元培养已成为研究中枢神经系统（CNS）发育的标准模型。此类培养主要从啮齿动物（小鼠和大鼠）的海马体或皮质制备，而其他哺乳动物较少使用。在此，我们描述了源自灰色南美短尾负鼠皮质的原代解离神经元培养的建立及广泛表征。负鼠在出生后早期发育阶段受伤后能够完全再生其CNS，这一能力使其独具特色。因此，我们使用出生后第3 - 5天（P3 - 5）负鼠的皮质来建立长期存活且近乎纯净的神经元培养物，以及由放射状胶质细胞（RGCs）组成的混合培养物，其中RGCs的神经发生和胶质发生潜能得到了证实。这两种类型的培养物都能存活超过1个月。我们还从P16 - 18负鼠皮质制备了神经元培养物，除神经元外，其还由星形胶质细胞和小胶质细胞组成。长期存活的负鼠原代解离神经元培养物代表了一个新型的哺乳动物平台，对研究CNS发育和再生特别有用。
