Hui Ling Ma, Raiane de Oliveira Ferreira, Danyllo Felipe de Oliveira, Alice Kei Endo, Oswaldo Keith Okamoto, Mayana Zatz
{"title":"3D生物打印神经祖细胞构建增强离体脑整合和神经元-星形胶质细胞分化","authors":"Hui Ling Ma, Raiane de Oliveira Ferreira, Danyllo Felipe de Oliveira, Alice Kei Endo, Oswaldo Keith Okamoto, Mayana Zatz","doi":"10.1016/j.bprint.2025.e00424","DOIUrl":null,"url":null,"abstract":"<div><div>Neural progenitor cells (NPCs) derived from human induced pluripotent stem cells (hiPSCs) hold great promise for neural tissue engineering, disease modeling, and regenerative therapies due to their self-renewal and differentiation potential. In this study, we utilized 3D extrusion bioprinting to encapsulate hiPSC-NPCs within a composite bioink composed of Gelatin methacryloyl (GelMA) and Pluronic F127 (P-127). This composite was engineered to enhance matrix remodeling, mechanical tunability, and cell-specific differentiation. Incorporating P-127 improved gelation, printability, swelling behavior, degradation kinetics, and microstructural features, collectively supporting enhanced NPC proliferation. Mechanical characterization revealed adjustable stiffness (Young's modulus: 1–8 kPa), with GelMA/P-127 blends exhibiting greater strength than GelMA alone. Immunostaining showed elevated GFAP and reduced Neurofilament M (NeuF-M) expression, indicating a shift toward astrocytic differentiation influenced by matrix mechanics. Calcium imaging and transient signal analysis confirmed the functional activity of the differentiated neurons. Gene expression profiling supported these findings, showing upregulation of GFAP, TUBB3, and MAP2 and downregulation of SOX2, marking the transition from progenitor to mature neural phenotypes. Furthermore, bioprinted constructs integrated with ex vivo brain slices and expressed TUBB3 and NeuF-M, confirming neuronal differentiation capacity. These results underscore the potential of GelMA/P-127 composite bioinks as biomimetic, tunable platforms for engineering 3D neural tissue constructs, offering a versatile tool for studying neurodevelopment and advancing translational regenerative strategies.</div></div>","PeriodicalId":37770,"journal":{"name":"Bioprinting","volume":"50 ","pages":"Article e00424"},"PeriodicalIF":0.0000,"publicationDate":"2025-07-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"3D bioprinted neural progenitor cell constructs for enhancing ex vivo brain integration and neuron-astrocyte differentiation\",\"authors\":\"Hui Ling Ma, Raiane de Oliveira Ferreira, Danyllo Felipe de Oliveira, Alice Kei Endo, Oswaldo Keith Okamoto, Mayana Zatz\",\"doi\":\"10.1016/j.bprint.2025.e00424\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<div><div>Neural progenitor cells (NPCs) derived from human induced pluripotent stem cells (hiPSCs) hold great promise for neural tissue engineering, disease modeling, and regenerative therapies due to their self-renewal and differentiation potential. In this study, we utilized 3D extrusion bioprinting to encapsulate hiPSC-NPCs within a composite bioink composed of Gelatin methacryloyl (GelMA) and Pluronic F127 (P-127). This composite was engineered to enhance matrix remodeling, mechanical tunability, and cell-specific differentiation. Incorporating P-127 improved gelation, printability, swelling behavior, degradation kinetics, and microstructural features, collectively supporting enhanced NPC proliferation. Mechanical characterization revealed adjustable stiffness (Young's modulus: 1–8 kPa), with GelMA/P-127 blends exhibiting greater strength than GelMA alone. Immunostaining showed elevated GFAP and reduced Neurofilament M (NeuF-M) expression, indicating a shift toward astrocytic differentiation influenced by matrix mechanics. Calcium imaging and transient signal analysis confirmed the functional activity of the differentiated neurons. Gene expression profiling supported these findings, showing upregulation of GFAP, TUBB3, and MAP2 and downregulation of SOX2, marking the transition from progenitor to mature neural phenotypes. Furthermore, bioprinted constructs integrated with ex vivo brain slices and expressed TUBB3 and NeuF-M, confirming neuronal differentiation capacity. These results underscore the potential of GelMA/P-127 composite bioinks as biomimetic, tunable platforms for engineering 3D neural tissue constructs, offering a versatile tool for studying neurodevelopment and advancing translational regenerative strategies.</div></div>\",\"PeriodicalId\":37770,\"journal\":{\"name\":\"Bioprinting\",\"volume\":\"50 \",\"pages\":\"Article e00424\"},\"PeriodicalIF\":0.0000,\"publicationDate\":\"2025-07-02\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Bioprinting\",\"FirstCategoryId\":\"1085\",\"ListUrlMain\":\"https://www.sciencedirect.com/science/article/pii/S2405886625000405\",\"RegionNum\":0,\"RegionCategory\":null,\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q1\",\"JCRName\":\"Computer Science\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Bioprinting","FirstCategoryId":"1085","ListUrlMain":"https://www.sciencedirect.com/science/article/pii/S2405886625000405","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q1","JCRName":"Computer Science","Score":null,"Total":0}
3D bioprinted neural progenitor cell constructs for enhancing ex vivo brain integration and neuron-astrocyte differentiation
Neural progenitor cells (NPCs) derived from human induced pluripotent stem cells (hiPSCs) hold great promise for neural tissue engineering, disease modeling, and regenerative therapies due to their self-renewal and differentiation potential. In this study, we utilized 3D extrusion bioprinting to encapsulate hiPSC-NPCs within a composite bioink composed of Gelatin methacryloyl (GelMA) and Pluronic F127 (P-127). This composite was engineered to enhance matrix remodeling, mechanical tunability, and cell-specific differentiation. Incorporating P-127 improved gelation, printability, swelling behavior, degradation kinetics, and microstructural features, collectively supporting enhanced NPC proliferation. Mechanical characterization revealed adjustable stiffness (Young's modulus: 1–8 kPa), with GelMA/P-127 blends exhibiting greater strength than GelMA alone. Immunostaining showed elevated GFAP and reduced Neurofilament M (NeuF-M) expression, indicating a shift toward astrocytic differentiation influenced by matrix mechanics. Calcium imaging and transient signal analysis confirmed the functional activity of the differentiated neurons. Gene expression profiling supported these findings, showing upregulation of GFAP, TUBB3, and MAP2 and downregulation of SOX2, marking the transition from progenitor to mature neural phenotypes. Furthermore, bioprinted constructs integrated with ex vivo brain slices and expressed TUBB3 and NeuF-M, confirming neuronal differentiation capacity. These results underscore the potential of GelMA/P-127 composite bioinks as biomimetic, tunable platforms for engineering 3D neural tissue constructs, offering a versatile tool for studying neurodevelopment and advancing translational regenerative strategies.
期刊介绍:
Bioprinting is a broad-spectrum, multidisciplinary journal that covers all aspects of 3D fabrication technology involving biological tissues, organs and cells for medical and biotechnology applications. Topics covered include nanomaterials, biomaterials, scaffolds, 3D printing technology, imaging and CAD/CAM software and hardware, post-printing bioreactor maturation, cell and biological factor patterning, biofabrication, tissue engineering and other applications of 3D bioprinting technology. Bioprinting publishes research reports describing novel results with high clinical significance in all areas of 3D bioprinting research. Bioprinting issues contain a wide variety of review and analysis articles covering topics relevant to 3D bioprinting ranging from basic biological, material and technical advances to pre-clinical and clinical applications of 3D bioprinting.