Perichondrocytes from microtia patients are a suitable cell source for tissue engineering of an auricle

Introduction

Tissue engineering is a promising strategy for the reconstruction of auricles for microtia patients. However, a high number of cells is required for a successful application of these methods. Since only small cell numbers can be isolated from the perichondrium, cells must be propagated over several passages in cell culture. However, little is known about the effects of cell culture on perichondrocytes (PCs). In this study, PCs from microtia patients were analysed with respect to cellular properties and chondrogenic markers at gene expression and protein level. The results obtained were compared with PCs from healthy donors.

Methods

Cells were isolated from perichondrium of microtia patients (n=3) and healthy donors (n=3) using outgrowth cell culture. (Ethical approval no: 2018-584N-MA) Colony forming and migration assays were performed to investigate general cell properties. Quantitative reverse transcriptase (qRT) PCR (Taqman assay) was used to analyse gene expression of cartilage-specific markers (SOX9, ACAN, COL1A1, and ELN). Flow cytometry was used to examine chondrogenic surface markers (CD90, CD44, CD73, CD29 and CD49e).

Results

The migratory capacity of PCs from microtia patients increased over the passages, in contrast to the cells from healthy donors. The ability to form colonies was not impaired. Specific markers for chondrocytes were constantly expressed. Surface markers specific for chondrogenic progenitor cells increased after isolation in the first passage and then remained constant.

Conclusions

PCs isolated from microtia patients retain their chondrogenic properties over four passages. This is sufficient to achieve a sufficient cell number for the engineering of an adult human auricle.

Supported by the SNF (Sinergia-Project 2-77120-17).

Schweizer Nationalfond (Sinergia-Projekt: 2-77120-17)

Publication History

Article published online:
24 May 2022