Bhavsar Rital B, Nakamura Kenta, Tsonis Panagiotis A
Department of Biology, University of Dayton.
J Vis Exp. 2011 Jun 22(52):2713. doi: 10.3791/2713.
Salamanders like newt and axolotl possess the ability to regenerate many of its lost body parts such as limbs, the tail with spinal cord, eye, brain, heart, the jaw¹. Specifically, newts are unique for its lens regeneration capability. Upon lens removal, IPE cells of the dorsal iris transdifferentiate to lens cells and eventually form a new lens in about a month²(,)³ . This property of regeneration is never exhibited by the ventral iris cells. The regeneration potential of the iris cells can be studied by making transplants of the in vitro cultured IPE cells. For the culture, the dorsal and ventral iris cells are first isolated from the eye and cultured separately for a time period of 2 weeks (Figure 1). These cultured cells are reaggregated and implanted back to the newt eye. Past studies have shown that the dorsal reaggregate maintains its lens forming capacity whereas the ventral aggregate does not form a lens, recapitulating, thus the in vivo process (Figure 2)⁴(,)⁵. This system of determining regeneration potential of dorsal and ventral iris cells is very useful in studying the role of genes and proteins involved in lens regeneration.
蝾螈和墨西哥钝口螈等蝾螈类动物具有再生许多缺失身体部位的能力，比如四肢、带有脊髓的尾巴、眼睛、大脑、心脏、颌骨¹。具体而言，蝾螈因其晶状体再生能力而独特。去除晶状体后，背侧虹膜的虹膜色素上皮（IPE）细胞会转分化为晶状体细胞，并最终在大约一个月内形成一个新的晶状体²（，）³。腹侧虹膜细胞则从不表现出这种再生特性。可以通过移植体外培养的IPE细胞来研究虹膜细胞的再生潜能。对于培养，首先从眼睛中分离出背侧和腹侧虹膜细胞，并分别培养两周（图1）。这些培养的细胞重新聚集后再植入蝾螈眼中。过去的研究表明，背侧细胞聚集体保持其形成晶状体的能力，而腹侧聚集体则不形成晶状体，从而重现了体内过程（图2）⁴（，）⁵。这种确定背侧和腹侧虹膜细胞再生潜能的系统在研究参与晶状体再生基因和蛋白质的作用方面非常有用。
