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基于多光子微加工和微图案化 (MMM) 的多重细胞生态位因子筛选用于表型维持——以牛髓核细胞为例
Biomaterials
(
IF
12.8
)
Pub Date : 2022-01-06
, DOI:
10.1016/j.biomaterials.2022.121367
Chi Hung Yip
1
,
Abigail Dee Chen
1
,
Yu Hin Wong
1
,
Barbara Pui Chan
1
Affiliation
- Tissue Engineering Laboratory, Biomedical Engineering Programme, Department of Mechanical Engineering, The University of Hong Kong, Pokfulam Road, Hong Kong.
在扁平和刚性塑料培养皿上进行单层培养后,许多细胞去分化并失去其天然表型。能够识别和重建最能维持培养物中生理细胞表型的细胞生态位因子的技术至关重要。我们开发了一种多光子微加工和微图案 (MMM) 技术,这是一种强大的 3D 微打印平台,能够制造蛋白质微结构和微图案,并对机械、拓扑和细胞外基质特性进行定量、空间和独立控制。在这里,以牛髓核细胞 (bNPCs) 为例,我们的目标是在体外重建一系列单个细胞生态位因子(2 个机械、9 个拓扑和 4 个基质),用于多重细胞生态位因子筛选,并制造一系列入围细胞生态位因子的最佳组合,以最好地维持 bNPC 表型。在筛选的所有因子中,两个拓扑(微柱阵列;纤维珠结构)和两个基质(层粘连蛋白;玻连蛋白)因子入围,从入围因子重组的组合细胞生态位因子被发现协同增强选定 bNPC 表型标记的表达(Col II、SNAP25 和角蛋白 8)并保持它们的形态和表型。这些最佳细胞壁龛可以微印在培养皿上,用于生理相关的培养,并有助于椎间盘组织工程的仿生支架设计。纤维珠结构)和两种基质(层粘连蛋白;玻连蛋白)因子入围,发现从入围因子重组的组合细胞生态位因子可协同增强选定 bNPC 表型标志物(Col II、SNAP25 和角蛋白 8)的表达并保持它们的形态和表型。这些最佳细胞壁龛可以微印在培养皿上,用于生理相关的培养,并有助于椎间盘组织工程的仿生支架设计。纤维珠结构)和两种基质(层粘连蛋白;玻连蛋白)因子入围,发现从入围因子重组的组合细胞生态位因子可协同增强选定 bNPC 表型标志物(Col II、SNAP25 和角蛋白 8)的表达并维持它们的形态和表型。这些最佳细胞壁龛可以微印在培养皿上,用于生理相关的培养,并有助于椎间盘组织工程的仿生支架设计。
"点击查看英文标题和摘要"
Multiphoton microfabrication and micropatternining (MMM)-based screening of multiplex cell niche factors for phenotype maintenance - Bovine nucleus pulposus cell as an example
Upon monolayer cultures on flat and rigid plastic dishes, many cells de-differentiate and lose their native phenotype. Technologies able to identify and reconstitute the cell niche factors that best maintain the physiological cellular phenotype in cultures are critical. We have developed a multiphoton microfabrication and micropatterning (MMM) technology, a robust 3D micro-printing platform capable to fabricate protein microstructures and micropatterns with quantitative, spatial and independent control of the mechanical, topological and extracellular matrix properties. Here, using bovine nucleus pulposus cells (bNPCs) as an example, we aim to reconstitute a spectrum of individual cell niche factors (2 mechanical, 9 topological and 4 matrices) in vitro for multiplex cell niche factor screening, and fabricate the optimal combinations of a series of shortlisted cell niche factors that best maintain the bNPC phenotype. Among all factors screened, two topological (micropillar array; fiber-bead structure) and two matrix (laminin; vitronectin) factors were shortlisted and the combinatory cell niche factors reconstituted from the shortlisted factors were found to synergistically augmented the expression of selected bNPC phenotype markers (Col II, SNAP25 and Keratin 8) and maintained their morphology and phenotype. These optimal cell niches can be micro-printed on culture dishes for physiologically relevant cultures and contribute to biomimetic scaffold design for intervertebral disc tissue engineering.
更新日期:2022-01-12