دورية أكاديمية
Use of l-pNIPAM hydrogel as a 3D-scaffold for intestinal crypts and stem cell tissue engineering.
| العنوان: | Use of l-pNIPAM hydrogel as a 3D-scaffold for intestinal crypts and stem cell tissue engineering. |
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| المؤلفون: | Dosh RH; Biomolecular Sciences Research Centre, Sheffield Hallam University, S1 1WB, UK. c.lemaitre@shu.ac.uk., Jordan-Mahy N, Sammon C, Le Maitre CL |
| المصدر: | Biomaterials science [Biomater Sci] 2019 Sep 24; Vol. 7 (10), pp. 4310-4324. |
| نوع المنشور: | Journal Article |
| اللغة: | English |
| بيانات الدورية: | Publisher: Royal Society of Chemistry Country of Publication: England NLM ID: 101593571 Publication Model: Print Cited Medium: Internet ISSN: 2047-4849 (Electronic) Linking ISSN: 20474830 NLM ISO Abbreviation: Biomater Sci Subsets: MEDLINE |
| أسماء مطبوعة: | Original Publication: Cambridge, UK : Royal Society of Chemistry |
| مواضيع طبية MeSH: | Stem Cells/*cytology , Tissue Engineering/*methods, Animals ; Caco-2 Cells ; Cell Differentiation/physiology ; Cell Line ; Humans ; Hydrogels ; Intestinal Mucosa/cytology ; Intestinal Mucosa/metabolism ; Intestine, Small/cytology ; Mice ; Mice, Inbred BALB C ; Receptors, G-Protein-Coupled/metabolism ; Stem Cells/metabolism |
| مستخلص: | Intestinal stem cells hold great potential in tissue regeneration of the intestine, however, there are key limitations in their culture in vitro. We previously reported a novel synthetic non-biodegradable hydrogel as a 3D culture model for intestinal epithelium using Caco2 and HT29-MTX cells. Here, we investigated the potential of this system as a 3D scaffold for crypts and single intestinal stem cells to support long-term culture and differentiation. Intestinal crypts were extracted from murine small intestines and Lgr5+ stem cells isolated by magnetic activated cell sorting. Crypts and stem cells were suspended within Matrigel or l-pNIPAM for 14 days or suspended within Matrigel for 7 days then released, dissociated, and suspended within, or on l-pNIPAM hydrogel for 28 days. Cellular behaviour and phenotype were determined by histology and immunohistochemistry for stem cell and differentiation markers: Lgr5, E-cadherin MUC2 chromograninA and lysozymes. Isolated crypts and Lgr5+ intestinal stem cells formed enteroids with a central lumen surrounded by multiple crypt-like buds when cultured in Matrigel. In contrast, when crypts and stem cells were directly suspended within, or layered on l-pNIPAM hydrogel under dynamic culture conditions they formed spherical balls of cells, with no central lumen. When enteroids were initially formed in Matrigel from crypts or single Lgr5+ intestinal stem cells and dissociated into small fragments or single cells and transferred to l-pNIPAM hydrogel they formed new larger enteroids with numerous crypt-like buds. These crypt-like buds showed the presence of mucin-producing cells, which resembled goblet cells, scattered throughout their structures. Immunohistochemistry staining also showed the expression of Lgr5 and differentiation markers of all the main intestinal cell types including: enterocytes, goblet cells, enteroendocrine and Paneth cells. This demonstrated that l-pNIPAM hydrogel supported long-term culture of crypts and Lgr5+ stem cells and promoted intestinal cell differentiation. |
| المشرفين على المادة: | 0 (Hydrogels) 0 (LGR5 protein, human) 0 (Receptors, G-Protein-Coupled) |
| تواريخ الأحداث: | Date Created: 20190815 Date Completed: 20200316 Latest Revision: 20200316 |
| رمز التحديث: | 20231215 |
| DOI: | 10.1039/c9bm00541b |
| PMID: | 31410428 |
| قاعدة البيانات: | MEDLINE |
| تدمد: | 2047-4849 |
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| DOI: | 10.1039/c9bm00541b |