Mesenchymal Stem Cells Isolated from Paediatric Paravertebral Adipose Tissue Show Strong Osteogenic Potential

Abstract

Mesenchymal stem cells (MSCs) represent the basis of novel clinical concepts in cellular therapy and tissue regeneration. Therefore, the isolation of MSCs from various tissues has become an important endeavour for stem cell biobanking and the development of regenerative therapies. Paravertebral adipose tissue is readily exposed during spinal procedures in children and could be a viable source of stem cells for therapeutic applications. Here, we describe the first case of MSCs isolated from paravertebral adipose tissue (PV-ADMSCs), obtained during a routine spinal surgery on a child. Using quantitative real-time PCR and flow cytometry, we show that PV-ADMSCs have different levels of stem marker expression compared to the MSCs from other sources while having the highest proliferation rate. Furthermore, we evaluate the multipotency of PV-ADMSCs by the three-lineage (adipogenic, osteogenic and chondrogenic) differentiation and compare it to the multipotency of MSCs from other sources. It was found that the PV-ADMSCs have a strong osteogenic potential in particular. Taken together, our data indicate that PV-ADMSCs meet the criteria for successful cell therapy, defined by the International Society for Cellular Therapy (ISCT), and thus, could provide a source of MSCs that is relatively easy to isolate and expand in culture. Due to their strong osteogenic potential, these cells provide a promising basis, especially for orthopaedic applications.

The Endplate Role in Degenerative Disc Disease Research: The Isolation of Human Chondrocytes from Vertebral Endplate—An Optimised Protocol

“…One of the important aspects to be considered is also the possibility that the cells that proliferate in the primary culture, belong to the pre-chondrocyte stem/progenitor population (i.e., chondroblasts) [63,64], which outgrows the isolated chondrocytes in a couple of passages. Furthermore, their phenotype is much better maintained in three-dimensional culture (e.g., pellet or hydrogel), where the cells are able to retain a somewhat spherical morphology [52,65]. Nevertheless, the obtained expression profiles indicate that despite the changes, the isolated cells retain some of their chondrocyte-specific characteristics even after the third passage, which is also in agreement with other literature sources [39,66].…”

“…During the isolation process, we noted that the best combination for fragment digestion included collagenase I and 5 wt.% FBS. We deduced this from our previous experimental practice [39,52]. We decided on a long digestion time of 19 h since this time was adequate to degrade the vertebral endplate fragments to a sufficient extent.…”