Gryshkov Oleksandr, Pogozhykh Denys, Hofmann Nicola, Pogozhykh Olena, Mueller Thomas, Glasmacher Birgit
Institute for Multiphase Processes, Leibniz Universitaet Hannover, Hannover, Germany.
Institute for Transfusion Medicine, Hannover Medical School, Hannover, Germany.
PLoS One. 2014 Sep 26;9(9):e107911. doi: 10.1371/journal.pone.0107911. eCollection 2014.
Alginate cell-based therapy requires further development focused on clinical application. To assess engraftment, risk of mutations and therapeutic benefit studies should be performed in an appropriate non-human primate model, such as the common marmoset (Callithrix jacchus). In this work we encapsulated amnion derived multipotent stromal cells (MSCs) from Callithrix jacchus in defined size alginate beads using a high voltage technique. Our results indicate that i) alginate-cell mixing procedure and cell concentration do not affect the diameter of alginate beads, ii) encapsulation of high cell numbers (up to 10×106 cells/ml) can be performed in alginate beads utilizing high voltage and iii) high voltage (15-30 kV) does not alter the viability, proliferation and differentiation capacity of MSCs post-encapsulation compared with alginate encapsulated cells produced by the traditional air-flow method. The consistent results were obtained over the period of 7 days of encapsulated MSCs culture and after cryopreservation utilizing a slow cooling procedure (1 K/min). The results of this work show that high voltage encapsulation can further be maximized to develop cell-based therapies with alginate beads in a non-human primate model towards human application.
基于藻酸盐的细胞疗法需要进一步发展，重点是临床应用。为了评估植入情况、突变风险和治疗效果，应在合适的非人灵长类动物模型中进行研究，例如普通狨猴（Callithrix jacchus）。在这项工作中，我们使用高压技术将来自普通狨猴的羊膜来源的多能间充质基质细胞（MSCs）封装在特定大小的藻酸盐珠中。我们的结果表明：i）藻酸盐与细胞的混合程序和细胞浓度不会影响藻酸盐珠的直径；ii）利用高压可在藻酸盐珠中封装高细胞数（高达10×106个细胞/毫升）；iii）与传统气流法生产的藻酸盐封装细胞相比，高压（15 - 30 kV）不会改变封装后MSCs的活力、增殖和分化能力。在封装的MSCs培养7天期间以及使用慢速冷却程序（1 K/分钟）进行冷冻保存后，均获得了一致的结果。这项工作的结果表明，高压封装可以进一步优化，以便在非人灵长类动物模型中开发基于藻酸盐珠的细胞疗法，从而应用于人类。
