The feasibility of the CD271+ and CD271– mesenchymal stromal cell enrichment toward nucleus pulposus-like cells

Abstract

introduction. Factors promoting nerve cell ingrowth are considered responsible for chronic back pain resulting from the intervertebral disc degeneration (IDD). One of the recent exploratory IDD treatments is stem cell transplantation therapy. The CD271 (low-affinity nerve growth factor receptor) has been identified as a marker of the most homogeneous mesenchymal stem cell (MSC) subset. It is capable of promoting differentiation along adipogenic, osteogenic and chondrogenic lineages and producing significantly higher levels of cytokines as compared to the total population of plastic adherence-mesenchymal stem cells (PA-MSCs). We investigated the ability of CD271 + MSCs to differentiate into chondrocyte-like cells of the nucleus pulposus (NP) of intervertebral disc. We also examined CD271 -MSCs, using PA-MSCs as a control cell population. Material and methods. Bone marrow derived PA-MSCs and its two subsets, CD271 -MSCs and CD271 + MSCs, were seeded in collagen scaffolds. After two weeks of growth in NP-differentiation medium, RNA was isolated from cells-scaffold constructs and was analyzed by q-PCR for expression of NP markers. Glycosaminoglycans were analyzed biochemically directly in cells-scaffold constructs. results. Expression of NP markers -extracellular matrix components such as aggrecan, collagen type II and glycosaminoglycans on both RNA and the protein levels -was significantly higher in CD271-MSCs compared to the CD271 + MSCs and PA-MSCs cell populations. Conclusions. CD271-MSCs may be superior candidates for NP restorative treatment compared to CD271 + MSCs and PA-MSCs due to their ability of expressing NP-supporting extracellular matrix components at levels higher than the other two studied MSC subsets.

Evaluation of regenerative processes in the pig model of intervertebral disc degeneration after transplantation of bone marrow-derived mesenchymal stem cells

“…The most straightforward approach to repair a degenerated disc is the delivery of different cells into the nucleus pulposus (NP) to increase extracellular matrix (ECM) synthesis. A variety of cells have been used, including NP cells [33][34][35], chondrocytes [1,19], and mesenchymal stem cells (MSCs) [13,[21][22][23]25,44,52,53]. All of these cell types have exhibited potential for slowing or repairing degeneration.…”

Controversies in regenerative medicine: Should intervertebral disc degeneration be treated with mesenchymal stem cells?

“…It recently became increasingly clear that preselection of particularly suitable MSC populations can further enhance the clinical outcome. CD271‐ MSCs, for example, have a higher potential for nucleus pulposus (NP)‐like differentiation than their CD271+ counterparts and may hence be primarily used in the future . In cartilage research, a shift towards subpopulations has already taken place.…”

“…Hence restoring GAG concentration is a requirement for disc repair. The reported amount of GAG produced by MSCs varies widely with species, cell source and culture conditions; in recent studies of MSCs in pellet or gel culture GAG production ranges from 0.017 to 0.086 mg GAG/million cells/month . GAG production rates increased more than 3‐fold by habituating MSCs to the harsh cellular microenvironment found in degenerate discs and through molecular biology techniques .…”

“…The reported amount of GAG produced by MSCs varies widely with species, cell source and culture conditions; in recent studies of MSCs in pellet or gel culture GAG production ranges from 0.017 to 0.086 mg GAG/million cells/month . GAG production rates increased more than 3‐fold by habituating MSCs to the harsh cellular microenvironment found in degenerate discs and through molecular biology techniques . Nevertheless, as the concentration of GAGs in the normal disc nucleus is around 70 mg/mL and since even the normal human lumbar disc cannot support more than around 3 million cells/mL, calculations show that it would take decades to restore the 25% of disc tissue (and hence of aggrecan) lost by the patient (Figure ) through implanting even stimulated MSCs.…”

Asperosaponin VI promotes human mesenchymal stem cell differentiation into nucleus pulposus like- cells via up-regulation of ERK1/2 and Smad2/3 signaling pathways.

“…Due to poor regeneration of NP cells, cell-based therapy may be a promising treatment for IVDD [7]. At present, increasing amounts of attention have been paid to using the multidirectional differentiation potential of stem cells to promote the regeneration of degenerative IVD [8].…”