Polymeric aqueous biphasic systems for non-contact cell printing on cells: engineering heterocellular embryonic stem cell niches.
Polymeric aqueous biphasic systems for non-contact cell printing on cells: engineering heterocellular embryonic stem cell niches.
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DOI：
10.1002/adma.200904271
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发表时间：
2010-06-25
影响因子：
29.4
通讯作者：
Takayama, Shuichi
中科院分区：
文献类型：
--
作者：
Tavana, Hossein;Mosadegh, Bobak;Takayama, Shuichi
通过将一种类型的细胞直接空间图案化到另一种细胞类型的活层上来产生异质细胞小生境的能力将有益于广泛的细胞生物学研究，包括干细胞研究。然而，大多数细胞图案化方法依赖于图案化的材料层来间接引导细胞图案化，[1-5]并且不能将细胞定位在已经存在的细胞单层上。细胞可以使用固体针直接嵌入到软支撑膜和凝胶中，[6，7]但所涉及的物理接触可能会损坏脆弱的基质，如活细胞。将单个细胞或细胞片轻轻置于细胞单层上，但在定位和分辨率控制方面存在局限性。[8-10]热或压电喷墨打印机利用热能或施加的电压从喷嘴喷射细胞悬浮液并将其导向基底。[11-13]虽然这种方法是非接触性的，并且可能适应多种细胞类型，但需要浓缩缓冲溶液[11]和特殊添加剂[14]，并且在细胞悬浮液制备和打印阶段仍然经常诱导化学，机械或热应力。此外，每种细胞类型都需要专用的盒，并且图案保真度是一个问题，因为细胞必须打印在浸入水性介质中的凝胶上。[14]压力介导的细胞接种水凝胶通过针头的挤出也产生了连续的细胞模式，[15]尽管由于凝胶包封而导致细胞与细胞的直接接触受损。因此，现有的方法面临的限制，在与活细胞和活细胞上的非接触式打印工程异细胞干细胞niches.We解决这个未满足的需求，通过使用聚合物水两相系统（ATPS），使非接触式打印高保真细胞图案到活细胞的精致表面在完全水性环境。高于一定浓度的聚乙二醇（PEG）和葡聚糖（DEX）的水溶液分离并形成ATPS，DEX始终形成底相（支持性信息，图S1）。[16]我们先前已经描述了PEG-DEX水性双相系统的效用，以有效地对细胞上的遗传物质进行基因过表达和RNA干扰基因沉默研究。[17]然而，由于细胞和基因递送构建体之间的物理化学差异，该制剂不能应用于细胞打印。在这里，我们定义了设计原则，以选择优化的ATPS配方，用于高效的细胞打印。使用这种方法，我们设计细胞小生境以支持mESC的神经元分化，并显示
The ability to generate heterocellular niches through direct spatial patterning of one type of cells onto a living layer of another cell type would benefit a wide range of cell biological studies including stem cell research. Most cell patterning approaches, however, rely on patterned material adhesiveness to indirectly guide cell patterning,[1–5] and cannot position cells on an already existing cell monolayer. Cells can be embedded directly into soft supported membranes and gels using solid pins,[6, 7] but the physical contact involved may damage delicate substrates such as living cells. Individual cells or cell sheets have been gently placed onto a cell monolayer, but with limitations in control of positioning and resolution.[8–10] Thermal or piezoelectric inkjet printers utilize thermal energy or applied electric voltage to eject the cell suspension from a nozzle and direct it toward the substrate.[11–13] Although this approach is non-contact and can potentially accommodate several cell types, concentrated buffer solutions [11] and special additives [14] are required and still often induce chemical, mechanical, or thermal stresses during cell suspension preparation and printing stages. Furthermore, dedicated cartridges are needed for each cell type, and pattern fidelity is an issue because cells have to be printed on a gel immersed in aqueous media.[14] Pressure-mediated extrusion of cell-seeded hydrogels through needles also creates continuous pattern of cells,[15] though with impaired direct cell-cell contact due to gel encapsulation. Therefore, existing approaches face limitations in terms of contact-free printing with and onto live cells for engineering heterocellular stem cell niches.We address this unmet need by using polymeric aqueous two-phase systems (ATPS) to enable non-contact printing of high-fidelity cellular patterns onto delicate surfaces of living cells in fully aqueous environments. Aqueous solutions of polyethylene glycol (PEG) and Dextran (DEX) above certain concentrations segregate and form an ATPS with DEX always forming the bottom phase (Supporting Information, Fig. S1).[16] We have previously described the utility of a PEG-DEX aqueous biphasic system to efficiently micropattern genetic materials on cells for gene overexpression and RNA interference gene silencing studies.[17] However due to physicochemical differences between cells and gene delivery constructs, that formulation could not be applied to cell printing. Here we define design principles to select optimized ATPS formulations for efficient cell printing. Using this approach, we engineer cellular niches to support neuronal differentiation of mESC and show
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影响因子：
14
作者：
Khademhosseini, A;Suh, KY;Langer, R
通讯作者：
Langer, R
影响因子：
2.9
作者：
TOUZET, N;SENSENBRENNER, M
通讯作者：
SENSENBRENNER, M
影响因子：
41.2
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影响因子：
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作者：
Roth, Therese M.;Ramamurthy, Poornapriya;Barald, Kate F.
通讯作者：
Barald, Kate F.
影响因子：
14
作者：
Rosenthal, Adam;Macdonald, Alice;Voldman, Joel
通讯作者：
Voldman, Joel