“…Cardiomyocyte differentiation from ESCs has been achieved in many species, including mice [12,21], monkeys [22,23], and humans [3,6]. The most frequently used protocol for differentiation involves EB formation [1, 9,[24][25][26][27][28][29][30][31][32][33].…”
Several approaches have been used to encourage the differentiation of cardiomyocytes from human embryonic stem cells. However, the differentiation efficiency is low, and appropriate culture protocols are needed to produce adequate numbers of cardiomyocytes for therapeutic cell transplantation. This study investigated the effects of serum on cardiomyocyte differentiation in suspension culture medium during embryoid body (EB) formation by human embryonic stem cells. The addition of ascorbic acid, dimethylsulfoxide and 5-aza-2'-deoxycytidine during days 5-7 at the EB-forming stage resulted in an increase in the numbers of rhythmically contracting clusters of derived cardiomyocytes. Treatment with 0.1 mmol L(-1) ascorbic acid alone, or more notably in combination with 10 micromol L(-1) 5-aza-2'-deoxycytidine, induced the formation of beating cells within EBs. Most of the beating clusters had spontaneous contraction rates similar to those found in human adults, and their contractile activity lasted for up to 194 days.
“…Cardiomyocyte differentiation from ESCs has been achieved in many species, including mice [12,21], monkeys [22,23], and humans [3,6]. The most frequently used protocol for differentiation involves EB formation [1, 9,[24][25][26][27][28][29][30][31][32][33].…”
Several approaches have been used to encourage the differentiation of cardiomyocytes from human embryonic stem cells. However, the differentiation efficiency is low, and appropriate culture protocols are needed to produce adequate numbers of cardiomyocytes for therapeutic cell transplantation. This study investigated the effects of serum on cardiomyocyte differentiation in suspension culture medium during embryoid body (EB) formation by human embryonic stem cells. The addition of ascorbic acid, dimethylsulfoxide and 5-aza-2'-deoxycytidine during days 5-7 at the EB-forming stage resulted in an increase in the numbers of rhythmically contracting clusters of derived cardiomyocytes. Treatment with 0.1 mmol L(-1) ascorbic acid alone, or more notably in combination with 10 micromol L(-1) 5-aza-2'-deoxycytidine, induced the formation of beating cells within EBs. Most of the beating clusters had spontaneous contraction rates similar to those found in human adults, and their contractile activity lasted for up to 194 days.
“…Recently, ES cell lines were established from other species such as rat [41] and monkey [42,43], and the derivation of cardiomyocytes from those ES cells described. Rat and monkey heart disease models could thus be used to evaluate functional improvements obtained following auto-transplantation of ES cell-derived cardiomyocytes, or the xenotransplantation of human ES/induced pluripotent stem (iPS) cell-derived cardiomyocytes.…”
Section: Cardiomyocyte Generation and Purification From Pluripotent Ementioning
“…Although BNP was originally discovered in the porcine brain, it is predominately produced from by the heart ventricles (Minamino et al, 1988; Abdelalim et al, 2006a,b). The physiological functions of BNP are induced by its binding to NP receptor-A (NPR-A; Misono et al, 2011).…”
Brain natriuretic peptide (BNP) exerts its functions through NP receptors. Recently, BNP has been shown to be involved in a wide range of functions. Previous studies reported BNP expression in the sensory afferent fibers in the dorsal horn (DH) of the spinal cord. However, BNP expression and function in the neurons of the central nervous system are still controversial. Therefore, in this study, we investigated BNP expression in the rat spinal cord in detail using reverse transcription-polymerase chain reaction (RT-PCR) and immunohistochemistry. RT-PCR analysis showed that BNP mRNA was present in the spinal cord and dorsal root ganglion (DRG). BNP immunoreactivity was observed in different structures of the spinal cord, including the neuronal cell bodies and neuronal processes. BNP immunoreactivity was observed in the DH of the spinal cord and in the neurons of the intermediate column (IC) and ventral horn (VH). Double-immunolabeling showed a high level of BNP expression in the afferent fibers (laminae I–II) labeled with calcitonin gene-related peptide (CGRP), suggesting BNP involvement in sensory function. In addition, BNP was co-localized with CGRP and choline acetyltransferase (ChAT) in the motor neurons of the VH. Together, these results indicate that BNP is expressed in sensory and motor systems of the spinal cord, suggesting its involvement in several biological actions on sensory and motor neurons via its binding to NP receptor-A (NPR-A) and/or NP receptor-B (NPR-B) at the spinal cord level.
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