Biophysik und Kryotechnologie, Fraunhofer IBMT, Sankt Ingbert, Germany.
PLoS One. 2013 Sep 9;8(9):e73498. doi: 10.1371/journal.pone.0073498. eCollection 2013.
The technique of immunoisolated transplantation has seen in the last twenty years improvements in biocompatibility, long term stability and methods for avoidance of fibrosis in alginate capsules. However, two major problems are not yet solved: living cellular material that is not centered in the capsule is not properly protected from the hosts' immune system and the total transplant volume needs to be reduced. To solve these problems, we present a method for applying fully biocompatible alginate multilayers to a barium-alginate core without the use of polycations. We report on the factors that influence layer formation and stability and can therefore provide data for full adjustability of the additional layer. Although known for yeast and plant cells, this technique has not previously been demonstrated with mammalian cells or ultra-high viscous alginates. Viability of murine insulinoma cells was investigated by live-dead staining and live cell imaging, for murine Langerhans' islets viability and insulin secretion have been measured. No hampering effects of the second alginate layer were found. This multi-layer technique therefore has great potential for clinical and in vitro use and is likely to be central in alginate matrix based immunoisolated cell therapy.
在过去的二十年中，免疫隔离移植技术在生物相容性、长期稳定性以及避免藻酸盐胶囊纤维化方面取得了进展。然而，仍有两个主要问题尚未解决：未位于胶囊中心的活体细胞材料不能很好地免受宿主免疫系统的攻击，并且需要减少总移植体积。为了解决这些问题，我们提出了一种在不使用聚阳离子的情况下将完全生物相容的藻酸盐多层应用于钡藻酸盐核心的方法。我们报告了影响层形成和稳定性的因素，因此可以为附加层的完全可调性提供数据。尽管这种技术已为酵母和植物细胞所熟知，但以前尚未在哺乳动物细胞或超高粘性藻酸盐中得到证明。通过死活染色和活细胞成像研究了鼠胰岛素瘤细胞的活力，并且已经测量了鼠朗格汉斯胰岛的活力和胰岛素分泌。没有发现第二层藻酸盐的阻碍作用。因此，这种多层技术具有很大的临床和体外应用潜力，并且可能是基于藻酸盐基质的免疫隔离细胞治疗的核心。
