Losino Noelia, Waisman Ariel, Solari Claudia, Luzzani Carlos, Espinosa Darío Fernández, Sassone Alina, Muro Andrés F, Miriuka Santiago, Sevlever Gustavo, Barañao Lino, Guberman Alejandra
Laboratorio de Regulación Génica en Células Madre, Departamento de Química Biológica, Facultad de Ciencias Exactas y Naturales, Universidad de Buenos Aires (UBA), Ciudad de Buenos Aires, Argentina ; Instituto de Química Biológica - Ciencias Exactas y Naturales (IQUIBICEN), UBA/Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Ciudad de Buenos Aires, Argentina.
PLoS One. 2013 Nov 14;8(11):e80681. doi: 10.1371/journal.pone.0080681. eCollection 2013.
Embryonic stem cells (ESC) need a set of specific factors to be propagated. They can also grow in conditioned medium (CM) derived from a bovine granulosa cell line BGC (BGC-CM), a medium that not only preserves their main features but also increases ESC´s proliferation rate. The mitogenic properties of this medium were previously reported, ascribing this effect to an alternative spliced generated fibronectin isoform that contains the extra domain A (FN EDA(+)). Here, we investigated if the FN EDA(+) isoform increased proliferation of mouse and human ES cells. We analyzed cell proliferation using conditioned media produced by different mouse embryonic fibroblast (MEF) lines genetically engineered to express FN constitutively including or excluding the EDA domain (FN EDA(-)), and in media supplemented with recombinant peptides containing or not the EDA. We found that the presence of EDA in the medium increased mouse and human ESC's proliferation rate. Here we showed for the first time that this FN isoform enhances ESC's proliferation. These findings suggest a possible conserved behavior for regulation of ES cells proliferation by this FN isoform and could contribute to improve their culturing conditions both for research and cell therapy.
胚胎干细胞（ESC）需要一组特定因子来进行增殖。它们也能在源自牛颗粒细胞系BGC的条件培养基（CM）（BGC-CM）中生长，这种培养基不仅能保持其主要特性，还能提高ESC的增殖率。此前已报道了这种培养基的促有丝分裂特性，将这种效应归因于一种包含额外A结构域（FN EDA(+)）的可变剪接产生的纤连蛋白异构体。在此，我们研究了FN EDA(+)异构体是否能提高小鼠和人类ES细胞的增殖。我们使用由不同的经基因工程改造以组成型表达FN（包括或不包括EDA结构域（FN EDA(-)））的小鼠胚胎成纤维细胞（MEF）系产生的条件培养基，以及补充有含或不含EDA的重组肽的培养基来分析细胞增殖。我们发现培养基中EDA的存在提高了小鼠和人类ESC的增殖率。在此我们首次表明这种FN异构体增强了ESC的增殖。这些发现提示了这种FN异构体在调节ES细胞增殖方面可能存在保守行为，并且可能有助于改善其用于研究和细胞治疗的培养条件。
