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Airway basal cells from human-induced pluripotent stem cells: a new frontier in cystic fibrosis research.
Demchenko, Anna; Belova, Lyubava; Balyasin, Maxim; Kochergin-Nikitsky, Konstantin; Kondrateva, Ekaterina; Voronina, Ekaterina; Pozhitnova, Victoria; Tabakov, Vyacheslav; Salikhova, Diana; Bukharova, Tatiana; Goldshtein, Dmitry; Kondratyeva, Elena; Kyian, Tatiana; Amelina, Elena; Zubkova, Olga; Popova, Olga; Ozharovskaia, Tatiana; Lavrov, Alexander; Smirnikhina, Svetlana.
Afiliação
  • Demchenko A; Laboratory of Genome Editing, Research Centre for Medical Genetics, Moscow, Russia.
  • Belova L; Laboratory of Genome Editing, Research Centre for Medical Genetics, Moscow, Russia.
  • Balyasin M; Scientific and Educational Resource Center, Peoples' Friendship University of Russia, Moscow, Russia.
  • Kochergin-Nikitsky K; Department of Cell Technology, Endocrinology Research Center, Moscow, Russia.
  • Kondrateva E; Laboratory of Genome Editing, Research Centre for Medical Genetics, Moscow, Russia.
  • Voronina E; Laboratory of Genome Editing, Research Centre for Medical Genetics, Moscow, Russia.
  • Pozhitnova V; Laboratory of Mutagenesis, Research Centre for Medical Genetics, Moscow, Russia.
  • Tabakov V; Laboratory of Mutagenesis, Research Centre for Medical Genetics, Moscow, Russia.
  • Salikhova D; Moscow Branch of the Biobank "All-Russian Collection of Biological Samples of Hereditary Diseases", Research Centre for Medical Genetics, Moscow, Russia.
  • Bukharova T; Stem Cell Genetics Laboratory, Research Centre for Medical Genetics, Moscow, Russia.
  • Goldshtein D; Stem Cell Genetics Laboratory, Research Centre for Medical Genetics, Moscow, Russia.
  • Kondratyeva E; Stem Cell Genetics Laboratory, Research Centre for Medical Genetics, Moscow, Russia.
  • Kyian T; Scientific and Clinical Department of Cystic Fibrosis, Research Centre for Medical Genetics, Moscow, Russia.
  • Amelina E; Scientific and Clinical Department of Cystic Fibrosis, Research Centre for Medical Genetics, Moscow, Russia.
  • Zubkova O; Laboratory of Cystic Fibrosis, Research Institute of Pulmonology, Moscow, Russia.
  • Popova O; Federal State Budget Institution "National Research Centre for Epidemiology and Microbiology Named After Honorary Academician N F Gamaleya" of the Ministry of Health of the Russian Federation, Moscow, Russia.
  • Ozharovskaia T; Federal State Budget Institution "National Research Centre for Epidemiology and Microbiology Named After Honorary Academician N F Gamaleya" of the Ministry of Health of the Russian Federation, Moscow, Russia.
  • Lavrov A; Federal State Budget Institution "National Research Centre for Epidemiology and Microbiology Named After Honorary Academician N F Gamaleya" of the Ministry of Health of the Russian Federation, Moscow, Russia.
  • Smirnikhina S; Laboratory of Genome Editing, Research Centre for Medical Genetics, Moscow, Russia.
Front Cell Dev Biol ; 12: 1336392, 2024.
Article em En | MEDLINE | ID: mdl-38737127
ABSTRACT
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.
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Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Idioma: En Revista: Front Cell Dev Biol Ano de publicação: 2024 Tipo de documento: Article

Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Idioma: En Revista: Front Cell Dev Biol Ano de publicação: 2024 Tipo de documento: Article