Department of Materials Science and Engineering, Graduate School of Engineering Science, Osaka University, 1-3 Machikaneyama-cho, Toyonaka, Osaka 560-8531, Japan.
Department of Materials Science and Engineering, Graduate School of Engineering Science, Osaka University, 1-3 Machikaneyama-cho, Toyonaka, Osaka 560-8531, Japan.
Biomaterials. 2015;53:494-501. doi: 10.1016/j.biomaterials.2015.02.119. Epub 2015 Mar 21.
Selective encapsulation of a particular cell population from heterogeneous cell populations has potential applications such as studies in cell-to-cell communication, regenerative medicine, and cell therapies. However, there are no versatile methods for realizing this. Here we report a method based on biospecific identification of the target cells through antigen-antibody reaction and subsequent enzymatic hydrogel sheath formation on the cell surfaces by horseradish peroxidase (HRP). Human hepatoma cell line HepG2 cells were selectively encapsulated in alginate-based hydrogel sheath from the mixture with mouse embryo fibroblast-like cell line 10T1/2 fibroblasts using anti-human CD326 antibody conjugated with HRP. The viability of the encapsulated cells was 93%. The cells released at 6 days of the encapsulation by degrading the sheath using alginate lyase grew almost the same as those free from encapsulation. The versatility of the method was confirmed using another antibody, cells, and hydrogel sheath material: Only human vein endothelial cells were encapsulated in gelatin-based hydrogel sheath from the mixture with 10T1/2 fibroblasts using anti-human CD31 antibody conjugated with HRP. The cell-selective encapsulation was also achieved by a system using a primary antibody with a secondary antibody conjugated with HRP.
从异质细胞群中选择性地包裹特定的细胞群体具有广泛的应用,例如在细胞间通讯、再生医学和细胞治疗等领域的研究。然而,目前还没有通用的方法来实现这一目标。在这里,我们报告了一种基于生物特异性识别靶细胞的方法,通过抗原-抗体反应,随后通过辣根过氧化物酶(HRP)在细胞表面形成酶促水凝胶鞘。用人抗 CD326 抗体与 HRP 偶联,从与人肝癌细胞系 HepG2 细胞和小鼠胚胎成纤维样细胞系 10T1/2 纤维母细胞的混合物中选择性地包裹 HepG2 细胞于藻酸盐基水凝胶鞘内。包封细胞的活力为 93%。通过用藻酸盐裂解酶降解鞘来释放包封 6 天的细胞,其生长情况与未包封的细胞几乎相同。该方法的通用性通过另一种抗体、细胞和水凝胶鞘材料得到了证实:用人抗 CD31 抗体与 HRP 偶联,从与人静脉内皮细胞和 10T1/2 纤维母细胞的混合物中选择性地包裹人静脉内皮细胞于明胶基水凝胶鞘内。该系统还使用与 HRP 偶联的二级抗体的一级抗体实现了细胞的选择性包裹。