Functional neural differentiation of human adipose tissue-derived stem cells using bFGF and forskolin

Abstract

BackgroundAdult mesenchymal stem cells (MSCs) derived from adipose tissue have the capacity to differentiate into mesenchymal as well as endodermal and ectodermal cell lineage in vitro. We characterized the multipotent ability of human adipose tissue-derived stem cells (hADSCs) as MSCs and investigated the neural differentiation potential of these cells.ResultsHuman ADSCs from earlobe fat maintained self-renewing capacity and differentiated into adipocytes, osteoblasts, or chondrocytes under specific culture conditions. Following neural induction with bFGF and forskolin, hADSCs were differentiated into various types of neural cells including neurons and glia in vitro. In neural differentiated-hADSCs (NI-hADSCs), the immunoreactivities for neural stem cell marker (nestin), neuronal markers (Tuj1, MAP2, NFL, NFM, NFH, NSE, and NeuN), astrocyte marker (GFAP), and oligodendrocyte marker (CNPase) were significantly increased than in the primary hADSCs. RT-PCR analysis demonstrated that the mRNA levels encoding for ABCG2, nestin, Tuj1, MAP2, NFL, NFM, NSE, GAP43, SNAP25, GFAP, and CNPase were also highly increased in NI-hADSCs. Moreover, NI-hADSCs acquired neuron-like functions characterized by the display of voltage-dependent tetrodotoxin (TTX)-sensitive sodium currents, outward potassium currents, and prominent negative resting membrane potentials under whole-cell patch clamp recordings. Further examination by RT-PCR showed that NI-hADSCs expressed high level of ionic channel genes for sodium (SCN5A), potassium (MaxiK, Kv4.2, and EAG2), and calcium channels (CACNA1C and CACNA1G), which were expressed constitutively in the primary hADSCs. In addition, we demonstrated that Kv4.3 and Eag1, potassium channel genes, and NE-Na, a TTX-sensitive sodium channel gene, were highly induced following neural differentiation.ConclusionsThese combined results indicate that hADSCs have the same self-renewing capacity and multipotency as stem cells, and can be differentiated into functional neurons using bFGF and forskolin.

Isolation and characterization of multipotent mesenchymal stem cells in nasal polyps

“…Neurofilaments are intermediate filaments found in neurons and they are believed to function primarily to provide structural support for axons. 29 In our study, the expression levels of NF-L, NF-H, and TuJ1 were significantly increased in npMSCs that had differentiated into neurogenic cells. This result suggests that npMSCs have the potential to differentiate into neuronlike cells.…”

Human muscle–derived stem/progenitor cells promote functional murine peripheral nerve regeneration

“…Adult stem cells capable of adopting the neural and/or glial phenotypes in vitro can be isolated from murine or human CNS (9)(10)(11), bone marrow (12)(13)(14)(15), umbilical cord blood (16)(17)(18), skin (19), hair follicles (20)(21)(22), adipose tissue (23)(24)(25)(26)(27)(28)(29), or dental pulp (30,31). Stem/progenitor cells isolated from murine and human skeletal muscles by various methods give rise to progeny cells with neuronal and glial phenotypes (12,(32)(33)(34)(35)(36).…”

The role of adipose derived stem cells, smooth muscle cells and low intensity laser irradiation (LILI) in tissue engineering and regenerative medicine

“…These cells have also shown increased viability and proliferation when exposed to LILI alone [5] and, as well as, in combination with epidermal growth factor (EGF) [28]. Recent studies have demonstrated the functional neural differentiation of human ADSCs [29]. Adipose stem cell treatments have been used in animals without adverse effects [30] and several clinical trials are underway in humans that have shown no adverse effects to date [31].…”

“…By 2007, in France, 32% of the deaths were caused by cardiovascular disease [2]. Since stem cells have the ability to self renew and differentiate into multiple lineages [3], these cells could be used in cellbased therapy for repairing and regenerating various tissues and organs. Many studies have been done involving LILI and stem cells, both in vitro and in vivo.…”