Mesenchymal stem cells from rat olfactory bulbs can differentiate into cells with cardiomyocyte characteristics

Abstract

Mesenchymal stromal/stem cells (MSCs) are widely distributed in different tissues such as bone marrow, adipose tissues, peripheral blood, umbilical cord and amnionic fluid. Recently, MSC-like cells were also found to exist in rat olfactory bulb and are capable of inducing differentiation into mesenchymal lineages - osteocytes, chondrocytes and adipocytes. However, whether these cells can differentiate into myocardial cells is not known. In this study, we examined whether olfactory bulb-derived MSCs could differentiate into myocardial cells in vitro. Fibroblast-like cells isolated from the olfactory bulb of neonatal rats were grown under four conditions: no treatment; in the presence of growth factors (neuregulin-1, bFGF and forskolin); co-cultured with cardiomyocytes; and co-cultured with cardiomyocytes plus neuregulin-1, bFGF and forskolin. Cell differentiation into myocardial cells was monitored by RT-PCR, light microscopy immunofluorescence, western blot analysis and contractile response to pharmacological treatments. The isolated olfactory bulb-derived fibroblast-like cells expressed CD29, CD44, CD90, CD105, CD166 but not CD34 and CD45, consistent with the characteristics of MSCs. Long cylindical cells that spontaneously contracted were only observed following 7 days of co-culture of MSCs with rat cardiomyocytes plus neuregulin-1, bFGF and forskolin. RT-PCR and western blot analysis indicated that the cylindrical cells expressed myocardial markers, such as Nkx2.5, GATA4, sarcomeric α-actinin, cardiac troponin I, cardiac myosin heavy chain, atrial natriuretic peptide and connexin 43. They also contained sarcomeres and gap junction and were sensitive to pharmacological treatments (adrenal and cholinergic agonists and antagonists). These findings indicate that rat olfactory bulb-derived fibroblast-like cells with MSC characteristics can differentiate into myocardial-like cells.

Olfactory ecto-mesenchymal stem cells possess immunoregulatory function and suppress autoimmune arthritis

“…Olfactory MSCs could differentiate into cells with cardiomyocyte characteristics or cells with bone-specific bioactivity. 22,24 Recently, human MSCs isolated from the olfactory mucosa were found to have a promyelination effect that contributed to the repairment of spinal cord injury. 23 However, it was not well characterized whether OEMSCs possess any similar immunoregulatory properties as other MSCs and if so, whether adoptive transfer of OE-MSCs could suppress autoimmune pathogenesis in vivo.…”

Olfactory Mucosa Mesenchymal Stem Cells and Biomaterials: A New Combination to Regenerative Therapies after Peripheral Nerve Injury

“…They exhibit high mitotic activity when compared to the bone marrow MSCs (BM-MSCs) (nearly three times higher) and are able to self-renew in long-term cultures (over 15 weeks) by maintaining telomerase activity and lack apoptotic activity [79]. The olfactory mucosa itself is a great cell source since its renewal continues throughout life and OM-MSC potency is not even afected by the age of the donor [80]. Diferent from BM-MSCs, OM-MSCs promote CNS myelination and induce the diferentiation of neural stem cells into oligodendrocytes and oligodendrocyte maturation.…”

“…The described collection methods include a delicate discarding of the turbinates [82,88] or complete collection of the olfactory bulb [80], being the harvested material taken in balanced solutions containing antibiotics and antifungals [82]. The olfactory epithelium or olfactory bulb tissue sample is then cut into small pieces, digested with collagenase and DNase [80,82] or dispase [78] to separate the cells from their extracellular matrix.…”

“…The olfactory epithelium or olfactory bulb tissue sample is then cut into small pieces, digested with collagenase and DNase [80,82] or dispase [78] to separate the cells from their extracellular matrix. Then, they are cultured in lasks containing appropriate medium like Dulbecco's modiied Eagle's medium or HAM´s F-12 supplemented with fetal calf serum or fetal bovine serum, antibiotics, antifungals and growth factors [75].…”

“…Determination of the phenotypic markers is achieved through low cytometric analysis where single cell suspensions are stained with speciic luorochrome-conjugated antibodies [78,80], RT-PCR [80] or by immunocytochemistry [82]. In addition, the identiication of speciic proteins of these cells can be performed using immunoluorescence microscopy [88].…”