In vitro enhancement and functional characterization of neurite outgrowth by undifferentiated adipose-derived stem cells

Abstract

Adipose-derived stem cells (AScs) can easily be obtained and expanded in vitro for use in autologous cell therapy. Via their production of cytokines and neurotrophic factors, transplanted AScs provide neuroprotection, neovascularization and induction of axonal sprouting. However, the influencing mechanism of undifferentiated AScs on nerve regeneration is currently only partially understood. In the present study, undifferentiated AScs and cutaneous primary afferent dorsal root ganglion (dRG) neurons were co-cultured in order to investigate their interaction. AScs were isolated from adult rat fat tissue. The presence of characteristic stem cell markers was determined by flow cytometry in three subsequent passages. Adipogenic, osteogenic, chondrogenic and glial differentiation was performed in order to evaluate their differentiation capacity. A direct co-culture system with dRG cells was established to determine the effect of undifferentiated pluripotent AScs on neurite elongation. Neurite outgrowth, length and number was examined in the co-culture and compared with single-culture cells and cells stimulated with nerve growth factor (NGF). In ASc cultures, NGF expression was assessed by ELISA. The present results demonstrated that the specific mesenchymal stem cell surface markers cd44, cd73 and cd90 were detected in all three subsequent passages of the isolated ASCs. In accordance, ASc differentiation into adipogenic, osteogenic, chondrogenic and Schwann cell phenotype was conducted successfully. Neurite outgrowth of dRG neurons was enhanced following co-culture with AScs, resulting in increased neurite length after 24 h of cultivation. Furthermore, neurite outgrowth of dRG neurons was directed towards the undifferentiated ASc and direct cell-to-cell contact was observed. In summary, the results of the present study revealed an interaction between the two cell types with guidance of neurite growth towards the undifferentiated ASC. These findings suggest that the use of undifferentiated ASc optimizing tissue-engineered constructs may be promising for peripheral nerve repair.

Neuroinflammation in Primary Cultures of the Rat Spinal Dorsal Horn Is Attenuated in the Presence of Adipose Tissue–Derived Medicinal Signalling Cells (AdMSCs) in a Co-cultivation Model

“…Studies performed by other workgroups highlighted beneficial effects of MSCs on nerve regeneration and neurite outgrowth [ 45 , 46 ]. We therefore investigated possible effects of co-cultivation with AdMSCs on growth and morphology of SDH neurons by means of immunocytochemistry.…”

“…The use of inserts with a fine-pored, permeable membrane provides the opportunity to investigate effects of released soluble mediators by both cultures on each other without allowing direct contact [ 61 , 62 ]. Stem cells of different origins have the capacity to stimulate neurite outgrowth and, therefore, support nerve regeneration in neurons of the peripheral nervous system [ 46 , 63 , 64 ]. This effect was improved by using MSCs differentiated into a Schwann cell phenotype for primary neurons from dorsal root ganglia in vitro [ 45 ] or in an in vivo model of sciatic nerve transection [ 65 ].…”

In Vitro and In Vivo Effects of Nobiletin on DRG Neurite Elongation and Axon Growth after Sciatic Nerve Injury

“…In the field of peripheral nerve regeneration, neurite outgrowth of DRG neurons (DRGs) is a main indicator to predict peripheral axonal regrowth after injury. de Siqueira-Santos et al [27] suggested that extending sensory fibers of DRGs in vitro might be a regenerative property for promoting functional recovery after SNI, and Bucan et al [28] Figure 5. Nobiletin increased axonal elongation 2 weeks after SNI.…”

“…In the field of peripheral nerve regeneration, neurite outgrowth of DRG neurons (DRGs) is a main indicator to predict peripheral axonal regrowth after injury. de Siqueira-Santos et al [27] suggested that extending sensory fibers of DRGs in vitro might be a regenerative property for promoting functional recovery after SNI, and Bucan et al [28] reported that DRG neurite elongation activated by exosomes forms adipose derived mesenchymal stem cells closely involved in peripheral nerve regeneration. In the present study, nobiletin dose-dependently regulated neurite outgrowth of primary cultured DRGs after SNI and DRG neurite elongation was significantly facilitated at 50 and 100 µM of nobiletin compared to other dosages.…”

adipoSIGHT in Therapeutic Response: Consequences in Osteosarcoma Treatment

“…The adipogenic and osteogenic differentiation of ASCs were induced by treating the cells using the respective media cocktail. Approximately 1 × 10 5 cells/cm 2 (p = 3) were seeded in a culture plate and treated with adipogenic (αMEM containing 1 µM dexamethasone, 0.5 mM 1-methyl-3-isobutylxanthine, 1 ng/mL insulin, and 100 µM indomethacin) [10] and osteogenic (αMEM supplemented with 10 mM β-glycerol phosphate, 10 mM dexamethasone, and 50 µg/mL ascorbic acid) [9] media for 21 days.…”