Microfluidics generation of chitosan microgels containing glycerylphytate crosslinker for in situ human mesenchymal stem cells encapsulation.
Microfluidics generation of chitosan microgels containing glycerylphytate crosslinker for in situ human mesenchymal stem cells encapsulation.
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DOI：
10.1016/j.msec.2020.111716
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发表时间：
2021-01
期刊：
影响因子：
--
通讯作者：
San Román J
中科院分区：
文献类型：
--
作者：
Mora-Boza A;Mancipe Castro LM;Schneider RS;Han WM;García AJ;Vázquez-Lasa B;San Román J
人间充质干细胞（hMSCs）由于其多种有益特性，即通过免疫调节和分泌因子，成为细胞治疗的一个有吸引力的来源。微流体技术对于细胞包封特别有吸引力，因为它提供了一种快速和可重复的方法来生成控制大小的微凝胶，并同时包封细胞。在这里，我们报道了在微流控装置中使用抗氧化剂植酸甘油酯（G1Phy）化合物和三聚磷酸盐（TPP）作为离子交联剂（G1Phy:TPP-微凝胶）的组合制备了基于水溶性壳聚糖原位亲离子凝胶化的hmsc负载微载体。这些微凝胶的粒径分布均匀，平均直径为104±12 μm，略低于对照组（127±16 μm， tpp -微凝胶）。与对照tpp微凝胶相比，G1Phy在微凝胶中的存在可以长期维持细胞活力，并在不利条件下上调旁分泌因子的分泌。将经G1Phy: tpp微凝胶包封的hMSCs通过注射递送至免疫功能低下小鼠皮下间隙，递送过程与注射未包封细胞一样简单。注射后立即，在表达荧光素酶的微凝胶包封和未包封的hMSCs之间观察到等效的信号强度，表明微载体对初始细胞存活没有不利影响。通过生物发光信号推断，细胞持久性随时间呈指数下降，与tpp微凝胶和未包被细胞相比，G1Phy: tpp微凝胶的半衰期值相对较高。总的来说，研究结果表明，微流体生成的G1Phy: tpp微凝胶是支持hMSC存活和修复活性的有前途的微载体。
Human mesenchymal stem cells (hMSCs) are an attractive source for cell therapies because of their multiple beneficial properties, i.e. via immunomodulation and secretory factors. Microfluidics is particularly attractive for cell encapsulation since it provides a rapid and reproducible methodology for microgel generation of controlled size and simultaneous cell encapsulation. Here, we report the fabrication of hMSC-laden microcarriers based on in situ ionotropic gelation of water-soluble chitosan in a microfluidic device using a combination of an antioxidant glycerylphytate (G1Phy) compound and tripolyphosphate (TPP) as ionic crosslinkers (G1Phy:TPP-microgels). These microgels showed homogeneous size distributions providing an average diameter of 104±12 μm, somewhat lower than that of control (127±16 μm, TPP-microgels). The presence of G1Phy in microgels maintained cell viability over time and upregulated paracrine factor secretion under adverse conditions compared to control TPP-microgels. Encapsulated hMSCs in G1Phy:TPP-microgels were delivered to the subcutaneous space of immunocompromised mice via injection, and the delivery process was as simple as the injection of unencapsulated cells. Immediately post-injection, equivalent signal intensities were observed between luciferase-expressing microgel-encapsulated and unencapsulated hMSCs, demonstrating no adverse effects of the microcarrier on initial cell survival. Cell persistence, inferred by bioluminescence signal, decreased exponentially over time showing relatively higher half-life values for G1Phy:TPP-microgels compared to TPP-microgels and unencapsulated cells. In overall, results position the microfluidics generated G1Phy:TPP-microgels as a promising microcarrier for supporting hMSC survival and reparative activities.
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