Isolation of Multipotent Mesenchymal Stromal Cells from Equine Tissues: Does Collagenase Digestion Impact the Cell Quality?

Aims: Multipotent mesenchymal stromal cells (MSC) are a promising therapeutic tool for the treatment of orthopaedic conditions in sports horses. In recent years, solid tissues were shown to be a source of high-quality MSC with superior proliferation potential compared with MSC derived from bone marrow. However, the standard cell isolation procedure by collagenase digestion of the tissue may impact the cellular quality. Therefore, the aim of this study was to compare this standard procedure with a potentially milder procedure of cell isolation. 

Methods: Equine adipose tissue, tendon tissue and umbilical cord tissue were collected and used for MSC isolation. One part of each specimen was minced and subjected to collagenase I digestion, followed by centrifugation steps and seeding of the isolated cells on culture plates. Another equal part was cut in 1 cm pieces which were placed on the culture dishes and covered with medium, allowing the MSC to migrate out of the tissue. Subsequently, the MSC yield after the first cell harvest as well as basic MSC characteristics such as their proliferation, migration and trilineage differentiation potential were assessed and compared. [...]