Previous studies indicate that a conditioned medium (CM) from glial cells, such as Schwann cells (SCs) and Olfactory Ensheathing Cells (OECs) can promote a neural differentiation of Adipose-derived mesenchymal Stem Cells (ASCs). In the present study, the effects of Ghrelin (Ghre) were tested on ASCs, as this hormone is able to improve the growth and differentiation of cultured cells. In the central nervous system, Ghre has been mainly observed in specific populations of hypothalamic neurons. However, its effects involve other nervous structures, such as amygdala and hippocampus. ASCs were isolated and expanded after collagenase digestion and centrifugation of raw lipoaspirates. Different culture conditions were tested. ASCs grown in the basal medium and two samples, grown in OEC-CM or SC-CM were considered as controls. In three other corresponding ASC samples, Ghre (2 ?M) was added. At day 1, 3 and 6 of growth, cells were tested by immunocytochemistry to detect the expression of some neural markers, such as PGP 9.5, MAP2 and GFAP. Results showed that all these markers were overexpressed when ASCs were cultured in OEC- or SC-CM. The combination of CM and Ghre further improved these increases. On the other hand, the addition of Ghre alone produced weaker effects. In general, these effects were clearly visible already at day 1 and were more evident at day 6. Overall, it is confirmed that the use of a glial CM may induce a neural differentiation of ASCs. Although Ghre alone was not capable of pronounced effects, its addition facilitated neural ASC differentiation. This study provides information on the action of Ghre, being able to modulate cell growth and differentiation
Ghrelin influence on neural differentiation of Adipose-derived mesenchymal Stem Cells
2019
Abstract
Previous studies indicate that a conditioned medium (CM) from glial cells, such as Schwann cells (SCs) and Olfactory Ensheathing Cells (OECs) can promote a neural differentiation of Adipose-derived mesenchymal Stem Cells (ASCs). In the present study, the effects of Ghrelin (Ghre) were tested on ASCs, as this hormone is able to improve the growth and differentiation of cultured cells. In the central nervous system, Ghre has been mainly observed in specific populations of hypothalamic neurons. However, its effects involve other nervous structures, such as amygdala and hippocampus. ASCs were isolated and expanded after collagenase digestion and centrifugation of raw lipoaspirates. Different culture conditions were tested. ASCs grown in the basal medium and two samples, grown in OEC-CM or SC-CM were considered as controls. In three other corresponding ASC samples, Ghre (2 ?M) was added. At day 1, 3 and 6 of growth, cells were tested by immunocytochemistry to detect the expression of some neural markers, such as PGP 9.5, MAP2 and GFAP. Results showed that all these markers were overexpressed when ASCs were cultured in OEC- or SC-CM. The combination of CM and Ghre further improved these increases. On the other hand, the addition of Ghre alone produced weaker effects. In general, these effects were clearly visible already at day 1 and were more evident at day 6. Overall, it is confirmed that the use of a glial CM may induce a neural differentiation of ASCs. Although Ghre alone was not capable of pronounced effects, its addition facilitated neural ASC differentiation. This study provides information on the action of Ghre, being able to modulate cell growth and differentiationI documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.
