In Vitro Neuronal Induction of Adipose-Derived Stem Cells and their Fate after Transplantation into Injured Mouse Brain

Abstract

The effect of substances known as inducers of neuronal differentiation on cultured human and mouse adipose-derived mesenchymal stem cells (ASCs) and their fate after transplantation into the injured and ischemic mouse brains were studied. ASCs were isolated from the human and mouse adipose tissue. Inducers of neuronal differentiation included β-mercaptoethanol, glial cell line-derived neurotrophic factor (GNDF), brain-derived neurotrophic factor (BDNF), retinoic acid (RA), 5-azacytidine, as well as their combinations. Three days after the induction, the phenotype of the induced cells was analyzed using immunocytochemistry and real-time PCR assay for differential expression of specific genes. The induction efficiency was evaluated by the increased transcription of neuronal differentiation markers: nestin, β-III-tubulin (Tub-B), microtubule-associated protein 2 (MAP2), and neuron-specific enolase (ENO2). The expression of marker genes was tested by immunocytochemical analysis. ASC cultivation in the medium with RA or BDNF in combination with 5- azacytidine for a week increased the mRNA and protein levels of nestin, Tub-B, and ENO2. The transplantation of induced mouse ASCs into the mouse brain increased the lifespan of the cells relative to control uninduced cells and promoted their migration from the transplantation site to the recipient cerebral parenchyma. The transplantation of the induced cells into the mouse brain pre-exposed to endothelin- 1 promoted a more active cell migration into the surrounding ischemic brain tissue. Thus, ASC exposure to RA or BDNF in combination with 5-azacytidine elevated the transcription of the neuronal differentiation markers and improved the viability and integration of ASCs grafted into the mouse brain.

Human mesenchymal stem cells - current trends and future prospective

“…Many studies have shown that factors like insulin, retinoic acid, bFGF, EGF, valproic acid, BME and hydrocortisone support neuronal differentiation of AD-MSCs [ 93 , 94 ]. Glial cell line-derived neurotrophic growth factors (GNDF), brain-derived neurotrophic factors (BDNF), retinoic acid, 5-azacytidine, isobutylmethylxanthine (IBMX) and indomethacin enhanced the MSCs differentiation into mature neuronal cells [ 95 ]. Gangliosides are glycosphingolipids which interact with EGF receptor (EGFR) and enhance osteoblast formation.…”

The differentiation potential of adipose tissue-derived mesenchymal stem cells into cell lineage related to male germ cells

“…RA alone has usually been used to promote the differentiation of ESCs and MSCs into germ cells (Geijsen et al, 2004;Nayernia et al, 2006;Drusenheimer et al, 2007;Kerkis et al, 2007). However, RA has been used in association with other substances to induce neuronal differentiation from cultured mouse AT-MSCs (Bi et al, 2010;Pavlova et al, 2012).…”

Mesenchymal Stem Cells Beyond Regenerative Medicine

“…Following adherence to plastic in vitro or tissue engraftment in vivo, MSCs form colonies and (trans)differentiate into a myriad of cell lineages (Kuznetsov et al, 1997;Wang et al, 2012;Vonk et al, 2018). For this to occur, their microenvironment must contain multiple mitogenic or stimulating factors (Tontonoz et al, 1994;Sekiya et al, 2002;Lucarelli et al, 2003;Solchaga et al, 2005;Fontaine et al, 2008;Inada et al, 2008;Pavlova et al, 2012); be subjected to hypoxic conditions (Mohyeldin et al, 2010;Zhang et al, 2019); or scaffolded to closely mimic organ architecture or function (Ouyang et al, 2003;Ohgushi et al, 2005). However, a newer understanding of the regenerative abilities of MSCs in vivo later emerged, linking tissue regrowth not to MSC (trans)differentiation exclusively but rather to autocrine and paracrine signaling transduced through their communication with local stimuli (Crisostomo et al, 2008), growth factors (Hahn et al, 2008), and inflammatory mediators (Haynesworth et al, 1996).…”