Objective: To compare a co‐culture system with a conventional dispase‐dissociation method for obtaining functional human respiratory epithelial cells from the nasal turbinates for tissue engineering application.
Methods: Human respiratory epithelial cells were serially passaged using a co‐culture system and a conventional dispase‐dissociation technique. The growth kinetics and gene expression levels of the cultured respiratory epithelial cells were compared. Four genes were investigated, namely cytokeratin‐18, a marker for ciliated and secretory epithelial cells; cytokeratin‐14, a marker for basal epithelial cells; MKI67, a proliferation marker; and MUC5B, a marker for mucin secretion. Immunocytochemical analysis was performed using monoclonal antibodies against the high molecular‐weight cytokeratin 34βE12, cytokeratin 18, and MUC5A to investigate the protein expression from cultured respiratory epithelial cells.
Results: Respiratory epithelial cells cultured using both methods maintained polygonal morphology throughout the passages. At passage 1, co‐cultured respiratory epithelial showed a 2.6‐times higher growth rate compared to conventional dispase dissociation technique, and 7.8 times higher at passage 2. Better basal gene expression was observed by co‐cultured respiratory epithelial cells compared to dispase dissociated cells. Immunocytochemical analyses were positive for the respiratory epithelial cells cultured using both techniques.
Conclusion: Co‐culture system produced superior quality of cultured human respiratory epithelial cells from the nasal turbinates as compared to dispase dissociation technique.
Background
Brown adipocytes are known to promote energy expenditure and limit weight gain to combat obesity.
Averrhoa bilimbi
, locally called belimbing buluh (DBB), is mainly used as an ethnomedicine in the treatment of metabolic disorders including diabetes mellitus, hypertension and obesity. The present study aims to investigate the browning activity on white adipocytes by
A. bilimbi
leaf extract and to evaluate the potential mechanisms.
Methods
Ethanolic leaf extract of
A. bilimbi
was exposed to Myf5 lineage precursor cells to stimulate adipocyte differentiation. Protein expressions of brown adipocyte markers were determined through high content screening analysis and validated through western blotting. Mito Stress Test assay was conducted to evaluate the cellular oxygen consumption rate upon
A. bilimbi
treatment.
Results
A. bilimbi
ethanolic leaf extract exhibited an adipogenesis effect similar to a PPARgamma agonist. It also demonstrated brown adipocyte differentiation in myoblastic Myf5-positive precursor cells. Expression of UCP1 and PRDM16 were induced. The basal metabolic rate and respiratory capacity of mitochondria were increased upon
A. bilimbi
treatment.
Conclusions
The findings suggest that
Averrhoa bilimbi
ethanolic leaf extract induces adipocyte browning through PRDM16 activation and enhances mitochondria activity due to UCP1 up-regulation.
Electronic supplementary material
The online version of this article (10.1186/s12906-019-2640-3) contains supplementary material, which is available to authorized users.
scite is a Brooklyn-based organization that helps researchers better discover and understand research articles through Smart Citations–citations that display the context of the citation and describe whether the article provides supporting or contrasting evidence. scite is used by students and researchers from around the world and is funded in part by the National Science Foundation and the National Institute on Drug Abuse of the National Institutes of Health.