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Identificando marcadores de superfície celular da haste neural primária e células progenitoras por rotulagem metabólica de Sialoglycan
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1Brain and Spinal Cord Innovative Research Center of Tongji Hospital, School of Life Sciences and Technology,Tongji University and Frontier Science Research Center for Stem Cells of Ministry of Education, 2College of Chemistry and Molecular Engineering, Peking-Tsinghua Center for Life Sciences, Beijing National Laboratory for Molecular Sciences, Synthetic and Functional Biomolecules Center, and Key Laboratory of Bioorganic Chemistry and Molecular Engineering of Ministry of Education,Peking University
Chapters
- 00:05Title
- 01:16Preparation of Mouse Endothelial Culture in Permeable Support Inserts
- 01:55Set-up of Neural-endothelial Co-culture and Ac4ManNAz Labeling System
- 03:25Immunofluorescent Staining of Sialoglycoproteins in Expanded Primary NSPCs and Differentiated Neurons
- 05:25Purification of Sialoglycoproteins from Expanded Primary NSPCs and Differentiated Neurons
- 08:45Results: Identifying Cell Surface Markers by Metabolic Labeling of Sialoglycan
- 10:47Conclusion
Aqui apresentamos um protocolo que combina um sistema de cocultura neural-endotelial in vitro e incorporação metabólica de sialoglicano com grupos funcionais bioortogonais para expandir as células primárias de tronco neural e progenitoras e rotular sua superfície sialoglycoproteínas para a análise da imagem latente ou da massa-espectrometria de marcadores da superfície da pilha.
