| Autor: |
Anna Demchenko, Lyubava Belova, Maxim Balyasin, Konstantin Kochergin-Nikitsky, Ekaterina Kondrateva, Ekaterina Voronina, Victoria Pozhitnova, Vyacheslav Tabakov, Diana Salikhova, Tatiana Bukharova, Dmitry Goldshtein, Elena Kondratyeva, Tatiana Kyian, Elena Amelina, Olga Zubkova, Olga Popova, Tatiana Ozharovskaia, Alexander Lavrov, Svetlana Smirnikhina |
| Jazyk: |
angličtina |
| Rok vydání: |
2024 |
| Předmět: |
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| Zdroj: |
Frontiers in Cell and Developmental Biology, Vol 12 (2024) |
| Druh dokumentu: |
article |
| ISSN: |
2296-634X |
| DOI: |
10.3389/fcell.2024.1336392 |
| Popis: |
Human-induced airway basal cells (hiBCs) derived from human-induced pluripotent stem cells (hiPSCs) offer a promising cell model for studying lung diseases, regenerative medicine, and developing new gene therapy methods. We analyzed existing differentiation protocols and proposed our own protocol for obtaining hiBCs, which involves step-by-step differentiation of hiPSCs into definitive endoderm, anterior foregut endoderm, NKX2.1+ lung progenitors, and cultivation on basal cell medium with subsequent cell sorting using the surface marker CD271 (NGFR). We derived hiBCs from two healthy cell lines and three cell lines with cystic fibrosis (CF). The obtained hiBCs, expressing basal cell markers (NGFR, KRT5, and TP63), could differentiate into lung organoids (LOs). We demonstrated that LOs derived from hiBCs can assess cystic fibrosis transmembrane conductance regulator (CFTR) channel function using the forskolin-induced swelling (FIS) assay. We also carried out non-viral (electroporation) and viral (recombinant adeno-associated virus (rAAV)) serotypes 6 and 9 and recombinant adenovirus (rAdV) serotype 5 transgene delivery to hiBCs and showed that rAAV serotype 6 is most effective against hiBCs, potentially applicable for gene therapy research. |
| Databáze: |
Directory of Open Access Journals |
| Externí odkaz: |
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