Efficient generation of neural stem cell-like cells from adult human bone marrow stromal cells

Abstract

Clonogenic neural stem cells (NSCs) are self-renewing cells that maintain the capacity to differentiate into brainspecific cell types, and may also replace or repair diseased brain tissue. NSCs can be directly isolated from fetal or adult nervous tissue, or derived from embryonic stem cells. Here, we describe the efficient conversion of human adult bone marrow stromal cells (hMSC) into a neural stem celllike population (hmNSC, for human marrow-derived NSClike cells). These cells grow in neurosphere-like structures, express high levels of early neuroectodermal markers, such as the proneural genes NeuroD1, Neurog2, MSl1 as well as otx1 and nestin, but lose the characteristics of mesodermal stromal cells. In the presence of selected growth factors, hmNSCs can be differentiated into the three main neural phenotypes: astroglia, oligodendroglia and neurons. Clonal analysis demonstrates that individual hmNSCs are multipotent and retain the capacity to generate both glia and neurons. Our cell culture system provides a powerful tool for investigating the molecular mechanisms of neural differentiation in adult human NSCs. hmNSCs may therefore ultimately help to treat acute and chronic neurodegenerative diseases.

Membrane Properties of Neuron-Like Cells Generated from Adult Human Bone-Marrow-Derived Mesenchymal Stem Cells

“…A majority of the action potentials that we detected had small overshoots and brief afterhyperpolarizations. The shape of the action potentials recorded from the neuron-like cells resembled the immature action potentials that are commonly observed for developing human neurons [26] and neuron-like cells differentiating from stem cells isolated from both humans [14,25,28,30] or from rodents [29,32,33]. In addition, the patch clamp recordings were atypical for mature neurons in that the spikes were not blocked by the sodium channel inhibitor TTX at concentrations up to 10 mM, and we did not observe any repetitive spikes or spontaneous action potentials.…”

“…However, the density of the voltage-gated sodium channels is very low compared to mature neurons, suggesting that the undifferentiated hMeSCs are incapable of producing action potentials. Attempts to induce action potentials in hMeSCs using depolarizing current pulses have confirmed that undifferentiated MeSCs isolated from the adult BM of humans or rodents do not generate action potentials [14,21,25,[37][38][39][40].…”

“…The hMeSCs are self-renewable multipotent stem cells capable of differentiating into various mesenchymal lineages, including osteogenic, chondrogenic, adipogenic, and fibroblastic cells [6][7][8]. Several recent studies have shown that hMeSCs are also capable of being directly induced into neuron-and glia-like cells under experimentally defined culture conditions [9][10][11][12][13][14][15][16] and after transplantation into the CNS [15,17,18]. This direct induction of MeSCs to differentiate into neuronal-and glial-like phenotypes usually involves exposure to growth factors, potentially toxic chemical agents, and cocultures with neural cells.…”

“…The BM-derived hMeSCs can be converted into NSC-like cells that can subsequently differentiate into neuron-and glia-like cells in culture [13,14,24,25]. Recently, we succeeded in generating NSC-like cells by cultivating the isolated hMeSCs in a conditioned medium of human neural stem cells (hNSCs-CM) without using a coculture system, chemical treatment, or genetic modification.…”

HSV-1 amplicon viral vector-mediated gene transfer to human bone marrow-derived mesenchymal stem cells

“…Human bone marrow-derived mesenchymal stem cells (BM-hMSCs) have been demonstrated to differentiate in vitro [1][2][3][4][5][6][7] and in vivo into neurallike cells following transplantation into mouse brain. 8,9 Currently, BM-hMSCs are used for preclinical treatment in myocardial infarction 10,11 and neuronal injury.…”