The impact of cryopreservation in signature markers and immunomodulatory profile of tendon and ligament derived cells

Tendon and ligament (T/L) engineering strategies towards clinical practice have been challenged by a paucity of understanding in the identification and still poorly described characterization of cellular niches. Prospecting how resident cell populations behave in vitro, and how cryopreservation may influence T/ L-promoting factors, can provide insights into T/ L-cellular profiles for novel regenerative solutions. Therefore, we studied human T/ L-derived cells isolated from patellar tendons and cruciate ligaments as suitable cellular models to anticipate tendon and ligament niches responses for advanced strategies with predictive tenogenic and ligamentogenic value. Our results show that the crude populations isolated from tendon and ligament tissues hold a stem cell subset and share a similar behavior in terms of tenogenic/ligamentogenic commitment. Both T/ L-derived cells successfully undergo cryopreservation/thawing maintaining the tenogenic/ligamentogenic profiles. The major differences between cryopreserved and fresh populations were observed at the gene expression of MKX, SCX, and TNMD as well as at the protein levels of collagen type I and III, in which cells from tendon origin (hTDCs) evidence increased values in comparison to the ones from ligament (hLDCs, pâ
Funded by the ERC CoG MagTendon (No. 772817), Fundação para a Ciência e Tecnologia (FCT) Project MagTT PTDC/CTM-CTM/29930/2017 (POCI-01-0145-FEDER-29930), project Norte-01-0145-FEDER-02219015 supported by Norte Portugal Regional Operational Programme (NORTE 2020) and H2020 Twinning project Achilles No. 810850. Doctoral grant PD/BD/128089/2016 of A. Vinhas was provided by FCT. The authors thank Hospital da Prelada (Porto, Portugal) for providing ligament and tendon samples.