Cultivate Primary Nasal Epithelial Cells from Children and Reprogram into Induced Pluripotent Stem Cells

Abstract

Nasal epithelial cells (NECs) are the part of the airways that respond to air pollutants and are the first cells infected with respiratory viruses. They are also involved in many airway diseases through their innate immune response and interaction with immune and airway stromal cells. NECs are of particular interest for studies in children due to their accessibility during clinical visits. Human induced pluripotent stem cells (iPSCs) have been generated from multiple cell types and are a powerful tool for modeling human development and disease, as well as for their potential applications in regenerative medicine. This is the first protocol to lay out methods for successful generation of iPSCs from NECs derived from pediatric participants for research purposes. It describes how to obtain nasal epithelial cells from children, how to generate primary NEC cultures from these samples, and how to reprogram primary NECs into well-characterized iPSCs. Nasal mucosa samples are useful in epidemiological studies related to the effects of air pollution in children, and provide an important tool for studying airway disease. Primary nasal cells and iPSCs derived from them can be a tool for providing unlimited material for patient-specific research in diverse areas of airway epithelial biology, including asthma and COPD research. Video LinkThe video component of this article can be found at

Prioritizing chronic obstructive pulmonary disease (COPD) candidate genes in COPD-related networks

“…Dampening the “innate immune response” to smoking played a critical role in modifying pulmonary inflammation and lung remodeling, which might slow the progression of COPD [ 30 ]. Nasal epithelial cells are involved in many airway diseases, including asthma and COPD, through their “innate immune response” and interaction with immune and airway stromal cells [ 31 ]. Bush et al found that genes important in “lung development” and early wheezing were implicated in COPD [ 32 ].…”

Standardizing Harvesting Methods for Human Turbinate Mesenchymal Stem Cells from Surgical Tissue

“…Being exposed, the olfactory nerve is a highly vulnerable organ and damage to this nerve results in hyposmia and anosmia in a large number of patients. Although hTMSCs are promising candidates for iPSC induction or as donor cells for such patients, only a few studies using hTMSCs have been reported [1,3]. An important road block currently faced is the low success rate of stem cell differentiation and iPSC induction.…”

Increased nasal epithelial cell responsiveness to IL‐17A in paediatric asthmatics with low blood neutrophil count, low traffic‐related air pollution exposure and good asthma control

“…3 Nasal epithelial cells (NECs) were collected from beneath the inferior turbinate using a cytology brush according to published techniques. 5 Importantly, cells were cultured for 1-4 weeks including two passages to limit effects of in vivo exposures (environmental, medications) on cellular readouts. To measure IL-17A responsiveness, passaged NECs were plated in 24-well plates, allowed to reach confluence and then treated with medium alone or IL-17A (100 ng/ml) for 18-20 h. Production of the IL-17A-induced chemokine, CXCL1 (GROα), was assessed in supernatants by ELISA.…”

“…Air pollutant exposure was estimated based on mean concentration of elemental carbon attributable to traffic (ECAT) exposure at birth and current address 3 . Nasal epithelial cells (NECs) were collected from beneath the inferior turbinate using a cytology brush according to published techniques 5 . Importantly, cells were cultured for 1–4 weeks including two passages to limit effects of in vivo exposures (environmental, medications) on cellular readouts.…”