Phenotypic characterization of regional human meniscus progenitor cells

Abstract

Stimulating meniscus regeneration using meniscal progenitor cells has been suggested as a promising new strategy. However, there is a lack of studies which decisively identify and characterize progenitor cell populations in human meniscus tissues. In this study, donor-matched progenitor cells were isolated via selective fibronectin adhesion from the avascular and vascular regions of the meniscus and chondroprogenitors from articular cartilage (n = 5). The mixed populations of cells from these regions were obtained by standard isolation techniques for comparison. The colony formation efficacy of avascular progenitors, vascular progenitors and chondroprogenitors was monitored using Cell-IQ® live cell imaging. Proliferation rates of progenitors were compared with their mixed population counterparts. Cell surface markers indicative of mesenchymal stromal cells profile and progenitor markers were characterized by flow cytometry in all populations. The fibrochondrogenic capacity was assessed via fibrochondrogenic differentiation and measuring GAG/DNA content and morphology. All meniscal progenitor and chondroprogenitor populations showed superior colony forming efficacy and faster proliferation rates compare to their mixed populations. Progenitor populations showed significantly higher positivity for CD49b and CD49c compared to their mixed population counterparts and chondroprogenitors had a higher positivity level of CD166 compared to mixed chondrocytes. GAG/DNA analysis demonstrated that progenitor cells generally produced more GAG than mixed populations. Our study demonstrates that the human meniscus contains meniscal progenitor populations in both the avascular and vascular regions. Meniscal progenitors derived from the vascular region exhibit enhanced proliferative and fibrochondrogenic characteristics compared to those from the avascular region; this may associate with the enhanced meniscal healing potential in the vascular region. These findings build on the body of evidence which suggests that meniscal progenitors represent an attractive cell therapy strategy for meniscal regeneration.

Allografts for partial meniscus repair: an in vitro and ex vivo meniscus culture study

“…Recent studies investigated the phenotype of meniscal cells using flow cytometry and single cell Frontiers in Bioengineering and Biotechnology frontiersin.org sequencing. They found regional differences with high amounts of CD90-positive cells and up to seven independent cell clusters (Sun et al, 2020;Wang et al, 2022). The low amount of CD146-positive cells in this study might indicate towards a special phenotype found in degenerated menisci: degenerated meniscus progenitor cells (DegP) (Sun et al, 2020).…”

In Vitro Safety and Efficacy Evaluation of a Juniperus communis Callus Culture Extract and Matricaria recutita Processing Waste Extract Combination as a Cosmetic Ingredient

“…Cell growth assessment was performed utilizing the Cell-IQ real-time cell monitoring platform (Chip-Man Technologies, Tampere, Finland) according to methodology adapted from the method described by Wang et al [ 72 ]. Then, 10 4 HaCaT keratinocytes were seeded into individual wells of a 24-well cell culture plate containing S10 medium (DMEM supplemented with 1% penicillin (100 U mL −1 )–streptomycin (100 μg mL −1 ) (P/S) and 10% fetal bovine serum (FBS, Sigma, St. Louis, MO, USA)).…”

A comparative study on various cell sources for constructing tissue-engineered meniscus

“…Scaffolds are essential in structural-oriented approaches to engineering meniscus tissues because they provide three-dimensional templates for cell growth and tissue formation ( Bilgen et al, 2018 ; Wang J et al, 2022 ). Structurally, the meniscus is featured by the unique gross crescent shape with hierarchical and intertwined fibrous ECM, which enables its good capability to resist complex loads in the knee joint ( Li et al, 2017 ).…”