Characteristics of Nasal Septal Cartilage–Derived Progenitor Cells during Prolonged Cultivation

Abstract

Objective To produce alternate cell sources for tissue regeneration, human nasal septal cartilage-derived progenitor cells (NSPs) were tested to identify whether these cells meet the criteria of cartilage progenitor cells. We also evaluated the effects of prolonged cultivation on the characteristics of NSPs. Study Design In vitro study. Setting Academic research laboratory. Methods NSPs were isolated from discarded human nasal septal cartilage. NSPs were cultured for 10 passages. The expression of septal progenitor cell surface markers was assessed by fluorescence-activated cell sorting. Cell proliferation was measured with a cell-counting kit. Cytokine secretion was analyzed with multiplex immunoassays. Chondrogenic differentiation of NSPs without differentiation induction was analyzed with type II collagen immunohistochemistry. Cartilage-specific protein expression was evaluated by Western blotting. Under osteo- and adipodifferentiation media, 2 lineage differentiation potentials were evaluated by histology and gene expression analysis. Results Surface epitope analysis revealed that NSPs are positive for mesenchymal stem cells markers and negative for hematopoietic cell markers. Cultured NSPs showed sufficient cell expansion and chondrogenic potential, as demonstrated by immunostaining and expression of cartilage-specific protein. IL-6, IL-8, and transforming growth factor ß were secreted by over 200 pg/mL. The osteo- and adipodifferentiation potentials of NSPs were identified by histology and specific gene expression. The aforementioned characteristics were not influenced by prolonged cultivation. Conclusion NSPs represent an initial step toward creating a cell source from surgically discarded tissue that may prove useful in cartilage regeneration.

Toward tissue-engineering of nasal cartilages

“…Low-passage (up to P3) human nasal septal chondrocytes have shown the capability to produce neocartilage containing GAG and type II collagen [89,90,99,[105][106][107]. It has also been suggested that the superficial zone of septal cartilage contains a promising population of nasoseptal progenitor cells (NSPs) [108]. These cells are migratory and express surface markers, such as CD29, CD105, CD106, CD90, and CCD44, suggesting a state of differentiation between mesenchymal stem cells (MSCs) and chondrocytes [109].…”

“…NSPs have been shown to differentiate to chondrogenic and osteogenic, but not adipogenic lineages [88] and show a greater proliferation potential than bone marrow-and adipose-derived MSCs [110]. The surface markers and differentiation potential of NSPs have been reported to remain unchanged after 10 passages [108]. The use of both septal chondrocytes and NSPs are limited greatly by donor site morbidity and tissue availability.…”

Tissue engineering applications in otolaryngology—The state of translation

“…Heatmap representing the proportion of publications by year that utilized specific translational research methodologies from construct characterization to human trial to investigate nasal tissue engineering 8‐24,98‐106 …”

“…Utilization of human chondrocytes for cartilage regeneration has been a focus within nasal TE. Nasal septal chondrocytes, in particular, have been shown to have favorable proliferative capacity and chondrogenic potential compared to other chondrocyte sources in in vitro studies 23,24 . This tissue is easily acquired, either as remnants from septal surgery or as a biopsy of the septum with minimal donor site morbidity.…”

Human nasal septal chondrocytes (NSCs) preconditioned on NSC-derived matrix improve their chondrogenic potential

“…All the studies using nasal septal chondrocytes (NSCs) were conducted after written approval (HC13TISI0038) obtained from the Institutional Review Board of the Catholic Medical Center Clinical Research Coordinating Center. Human nasal septum tissue was harvested from ve patients who were undergoing septoplasty [21]. Those tissues were cut into 1 mm 3 pieces and they were enzymatically digested using 0.2% protease solution (Gibco) for 60 min, followed by incubation in 0.3% collagenase (Sigma-Aldrich) for 12hr at 37 °C.…”