Bond Wesley S, Wadhwa Lalita, Perlaky Laszlo, Penland Rebecca L, Hurwitz Mary Y, Hurwitz Richard L, Chèvez-Barrios Patricia
Interdepartmental Program in Translational Biology & Molecular Medicine, Baylor College of Medicine, USA.
J Vis Exp. 2011 Aug 4(54):2644. doi: 10.3791/2644.
Culturing retinoblastoma tumor cells in defined stem cell media gives rise to primary tumorspheres that can be grown and maintained for only a limited time. These cultured tumorspheres may exhibit markedly different cellular phenotypes when compared to the original tumors. Demonstration that cultured cells have the capability of forming new tumors is important to ensure that cultured cells model the biology of the original tumor. Here we present a protocol for propagating human retinoblastoma tumors in vivo using Rag2(-/-) immune deficient mice. Cultured human retinoblastoma tumorspheres of low passage or cells obtained from freshly harvested human retinoblastoma tumors injected directly into the vitreous cavity of murine eyes form tumors within 2-4 weeks. These tumors can be harvested and either further passaged into murine eyes in vivo or grown as tumorspheres in vitro. Propagation has been successfully carried out for at least three passages thus establishing a continuing source of human retinoblastoma tissue for further experimentation.
在特定的干细胞培养基中培养视网膜母细胞瘤肿瘤细胞会产生原发性肿瘤球，这些肿瘤球只能在有限的时间内生长和维持。与原始肿瘤相比，这些培养的肿瘤球可能表现出明显不同的细胞表型。证明培养的细胞具有形成新肿瘤的能力对于确保培养的细胞模拟原始肿瘤的生物学特性很重要。在这里，我们展示了一种使用Rag2(-/-)免疫缺陷小鼠在体内繁殖人类视网膜母细胞瘤肿瘤的方案。低传代的培养人类视网膜母细胞瘤肿瘤球或从新鲜收获的人类视网膜母细胞瘤肿瘤中获得的细胞直接注射到小鼠眼睛的玻璃体腔中，在2-4周内形成肿瘤。这些肿瘤可以被收获，然后在体内进一步接种到小鼠眼睛中，或者在体外作为肿瘤球生长。已经成功进行了至少三代的传代，从而建立了用于进一步实验的人类视网膜母细胞瘤组织的持续来源。
