scholarly journals Evaluation of the Neurogenic Potential in the Rat Inferior Colliculus from Early Postnatal Days Until Adulthood

2020 ◽  
Author(s):  
Jonas Engert ◽  
Kristen Rak ◽  
Linda Bieniussa ◽  
Miriam Scholl ◽  
Rudolf Hagen ◽  
...  
Keyword(s):  
Stem Cells ◽  
Glial Fibrillary Acidic Protein ◽  
Inferior Colliculus ◽  
Basic Protein ◽  
Single Cells ◽  
Auditory Pathway ◽  
Neural Lineage ◽  
Qrt Pcr ◽  
Histological Sections ◽  
Lineage Markers

Abstract Neural stem cells (NSCs) have been recently identified in the inferior colliculus (IC). These cells are of particular interest, as no casual therapeutic options for impaired neural structures exist. This research project aims to evaluate the neurogenic potential in the rat IC from early postnatal days until adulthood. The IC of rats from postnatal day 6 up to 48 was examined by neurosphere assays and histological sections. In free-floating IC cell cultures, neurospheres formed from animals from early postnatal to adulthood. The amount of generated neurospheres decreased in older ages and increased with the number of cell line passages. Cells in the neurospheres and the histological sections stained positively with NSC markers (Doublecortin, Sox-2, Musashi-1, Nestin, and Atoh1). Dissociated single cells from the neurospheres differentiated and were stained positively for the neural lineage markers β-III-tubulin, glial fibrillary acidic protein, and myelin basic protein. In addition, NSC markers (Doublecortin, Sox-2, CDK5R1, and Ascl-1) were investigated by qRT-PCR. In conclusion, a neurogenic potential in the rat IC was detected and evaluated from early postnatal days until adulthood. The identification of NSCs in the rat IC and their age-specific characteristics contribute to a better understanding of the development and the plasticity of the auditory pathway and might be activated for therapeutic use.

scholarly journals Isolation and Characterization of Neural Stem Cells from the Rat Inferior Colliculus

2019 ◽  
Vol 2019 ◽  
pp. 1-12 ◽  
Author(s):  
Johannes Völker ◽  
Jonas Engert ◽  
Christine Völker ◽  
Linda Bieniussa ◽  
Philipp Schendzielorz ◽  
...  
Keyword(s):  
Stem Cells ◽  
Stem Cell ◽  
Neural Stem Cells ◽  
Inferior Colliculus ◽  
Single Cells ◽  
Auditory Pathway ◽  
Superior Olivary Complex ◽  
Stem Cell Markers ◽  
Isolation And Characterization

The inferior colliculus (IC) is a nucleus of the auditory pathway and its fourth relay station. It integrates afferent information from the superior olivary complex and the cochlear nucleus. To date, no causal therapeutic options are known for damaged neuronal structures in this area. Regenerative medicine offers a potential approach to causally treating hearing impairment. After neural stem cells had been identified in certain areas of the auditory pathway, the question arouses, whether the IC also has a neurogenic potential. Cells from the IC of postnatal day 6 rats were extracted and cultured as neurospheres. Cells in the neurospheres showed mitotic activity and positive stain of neural stem cell markers (Nestin, DCX, Atoh1, and Sox-2). In addition, single cells were differentiated into neuronal and glial cells shown by the markers β-III-tubulin, GFAP, and MBP. In summary, basic stem cell criteria could be detected and characterized in cells isolated from the IC of the rat. These findings will lead to a better understanding of the development of the auditory pathway and may also be relevant for identifying causal therapeutic approaches in the future.

346 EXPRESSION OF IMPRINTED NONCODING RNA FROM THE DLK1-DIO3 LOCUS IN HUMAN EMBRYONIC STEM CELLS ADVANTAGES NEURAL LINEAGE DIFFERENTIATION

2015 ◽  
Vol 27 (1) ◽  
pp. 261
Author(s):  
C.-F. Mo ◽  
F.-C. Wu ◽  
K.-Y. T. Tai ◽  
W.-C. Chang ◽  
K.-W. Chang ◽  
...  
Keyword(s):  
Stem Cells ◽  
Noncoding Rna ◽  
Expression Profiles ◽  
Embryonic Stem ◽  
Gene Expression Profiles ◽  
Expression Levels ◽  
Neural Lineage ◽  
Qrt Pcr ◽  
Lineage Differentiation ◽  
Neurite Formation

Pluripotent stem cells are increasingly used for therapeutic models, including transplantation of neural progenitors derived from human embryonic stem cells (hESC). Recently, long noncoding RNA (lncRNA), including Maternally Expressed Gene 3 (MEG3) derived from the DLK1-DIO3-imprinted locus, were found to be expressed during neural developmental events. Their deregulations are associated with various neurological diseases. The DLK1-DIO3-imprinted locus encodes abundant noncoding RNA (ncRNA) that are regulated by differential methylation on the locus. The aim of our research was to study the correlation between the DLK1-DIO3-derived ncRNA and the capacity of hESC neural lineage differentiation. We classified hESC into MEG3-ON and MEG3-OFF based on the expression levels of MEG3 as well as its downstream microRNA by qRT-PCR. Initial embryoid body (EB) formation was conducted to examine the 3 germ layer's differentiation ability. Complementary DNA microarray was used to analyse the gene expression profiles of hESC. Directed neural lineage differentiation was performed, followed by analysis of neural lineage marker expression levels and neurite formation via qRT-PCR and immunocytochemistry methods to investigate the capacity of neural differentiation in MEG3-ON and MEG3-OFF hESC. As for statistics, error bars indicate standard error of the mean. Student's t-test was used for calculating P-values, and a P-value of less than 0.05 was considered to be significant. Our results showed that MEG3-ON and MEG3-OFF hESC differed greatly in DLK1-DIO3-derived ncRNA expression levels, but had comparable pluripotency gene expression profiles. Genes related to nervous system development and neural cancers were differentially expressed in MEG3-OFF hESC, where DLK1-DIO3-derived ncRNA were repressed compared to MEG3-ON ones before differentiation. In neural lineage-like cells derived from MEG3-OFF hESC, lower expression levels of neural lineage markers and impaired neurite formation were observed compared to MEG3-ON hESC at the same time points after differentiation. We suggest that the expression of DLK1-DIO3-derived lncRNA, MEG3, can be used as a simple and effective screening criterion for identifying MEG3-ON hESC with activation of DLK1-DIO3-imprinted ncRNA as starting materials to benefit neural lineage-associated studies.

Overexpression of Pygo2 Increases Differentiation of Human Umbilical Cord Mesenchymal Stem Cells into Cardiomyocyte-like Cells

2020 ◽  
Vol 20 (4) ◽  
pp. 318-324 ◽  
Author(s):  
Lei Yang ◽  
Shuoji Zhu ◽  
Yongqing Li ◽  
Jian Zhuang ◽  
Jimei Chen ◽  
...  
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Umbilical Cord ◽  
Heart Function ◽  
Critical Role ◽  
Human Umbilical Cord ◽  
Stem Cell Markers ◽  
Quantitative Real Time Pcr ◽  
Qrt Pcr

Background: Our previous studies have shown that Pygo (Pygopus) in Drosophila plays a critical role in adult heart function that is likely conserved in mammals. However, its role in the differentiation of human umbilical cord mesenchymal stem cells (hUC-MSCs) into cardiomyocytes remains unknown. Objective: To investigate the role of pygo2 in the differentiation of hUC-MSCs into cardiomyocytes. Methods: Third passage hUC-MSCs were divided into two groups: a p+ group infected with the GV492-pygo2 virus and a p− group infected with the GV492 virus. After infection and 3 or 21 days of incubation, Quantitative real-time PCR (qRT-PCR) was performed to detect pluripotency markers, including OCT-4 and SOX2. Nkx2.5, Gata-4 and cTnT were detected by immunofluorescence at 7, 14 and 21 days post-infection, respectively. Expression of cardiac-related genes—including Nkx2.5, Gata-4, TNNT2, MEF2c, ISL-1, FOXH1, KDR, αMHC and α-Actin—were analyzed by qRT-PCR following transfection with the virus at one, two and three weeks. Results : After three days of incubation, there were no significant changes in the expression of the pluripotency stem cell markers OCT-4 and SOX2 in the p+ group hUC-MSCs relative to controls (OCT-4: 1.03 ± 0.096 VS 1, P > 0.05, SOX2: 1.071 ± 0.189 VS 1, P > 0.05); however, after 21 days, significant decreases were observed (OCT-4: 0.164 ± 0.098 VS 1, P < 0.01, SOX2: 0.209 ± 0.109 VS 1, P < 0.001). Seven days following incubation, expression of mesoderm specialisation markers, such as Nkx2.5, Gata-4, MEF2c and KDR, were increased; at 14 days following incubation, expression of cardiac genes, such as Nkx2.5, Gata-4, TNNT2, MEF2c, ISL-1, FOXH1, KDR, αMHC and α-Actin, were significantly upregulated in the p+ group relative to the p− group (P < 0.05). Taken together, these findings suggest that overexpression of pygo2 results in more hUCMSCs gradually differentiating into cardiomyocyte-like cells. Conclusion: We are the first to show that overexpression of pygo2 significantly enhances the expression of cardiac-genic genes, including Nkx2.5 and Gata-4, and promotes the differentiation of hUC-MSCs into cardiomyocyte-like cells.

scholarly journals BAC Transgenesis in Human Embryonic Stem Cells as a Novel Tool to Define the Human Neural Lineage

Stem Cells ◽  
2009 ◽  
Vol 27 (3) ◽  
pp. 521-532 ◽  
Author(s):  
Dimitris G. Placantonakis ◽  
Mark J. Tomishima ◽  
Fabien Lafaille ◽  
Sabrina C. Desbordes ◽  
Fan Jia ◽  
...  
Keyword(s):  
Stem Cells ◽  
Embryonic Stem Cells ◽  
Human Embryonic Stem Cells ◽  
Embryonic Stem ◽  
Neural Lineage ◽  
Bac Transgenesis

scholarly journals miR-16-2* Interferes with WNT5A to Regulate Osteogenesis of Mesenchymal Stem Cells

2018 ◽  
Vol 51 (3) ◽  
pp. 1087-1102 ◽  
Author(s):  
Lijun Duan ◽  
He Zhao ◽  
Yang Xiong ◽  
Xiangsheng Tang ◽  
Yongdong Yang ◽  
...  
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Bone Formation ◽  
Osteoblast Differentiation ◽  
Target Genes ◽  
Fragility Fractures ◽  
Inhibitory Effect ◽  
Lymphocytic Leukemia ◽  
Qrt Pcr ◽  
Wnt Signal

Background/Aims: Osteoporosis is a bone metabolic disease characterized by a systemic impairment of bone mass, which results in increased propensity of fragility fractures. A reduction in the differentiation of MSCs into osteoblasts contributes to the impaired bone formation observed in osteoporosis. Mesenchymal stem cells (MSCs) are induced to differentiate into preosteoblasts, which are regulated by the signaling cascades initiated by the various signals, including miRNAs. miR-16-2* is a newly discovered miRNA that participates in diagnosis and prognosis of hepatocellular carcinoma, cervical cancer and chronic lymphocytic leukemia. However, the effect of miR-16-2* on the regulation of osteoblast differentiation and the mechanism responsible are still unclear. Here we discuss the contribution of miR-16-2* to osteoporosis, osteoblast differentiation and mineralization. Methods: The expression pattern of miR-16-2* during osteogenesis or in osteoporosis bone samples was validated by quantitative real-time PCR (qRT-PCR). The human bone marrow mesenchymal stem cells (hBMSCs) were induced to differentiate into osteoblasts by osteogenic induced medium containing dexamethasone, ascorbate-2-phosphat, beta-glycerophosphate and vitamin-D3. The target genes of miR-16-2* were predicted by TargetScan and PicTar. The mRNA and protein levels of osteogenic key markers were detected using qRT-PCR or western blot respectively. The WNT signal activity was analyzed by TOP/FOP reporter assay. Results: The expression of miR-16-2* in patient bone tissue with osteoporosis was negatively correlated with bone formation related genes. During osteoblast differentiation process, the expression of miR-16-2* was significantly decreased. Upregulation of miR-16-2* in hBMSCs impaired the osteogenic differentiation while the downregulation of miR-16-2* increased this process. Upregulation the expression of miR-16-2* could also block the WNT signal pathway by directly target WNT5A. Furthermore, knockdown of miR-16-2* could promote the activation of RUNX2, possibly by lifting the inhibitory effect of miR-16-2* on WNT pathway. Conclusion: Taken together, we report a novel biological role of miR-16-2* in osteogenesis through regulating WNT5A response for the first time. Our data support the potential utilization of miRNA-based therapies in regenerative medicine.

A Mesenchymal-Like ZEB1+Niche Harbors Dorsal Radial Glial Fibrillary Acidic Protein-Positive Stem Cells in the Spinal Cord

Stem Cells ◽  
2009 ◽  
Vol 27 (11) ◽  
pp. 2722-2733 ◽  
Author(s):  
Jean-Charles Sabourin ◽  
Karin B. Ackema ◽  
David Ohayon ◽  
Pierre-Olivier Guichet ◽  
Florence E. Perrin ◽  
...  
Keyword(s):  
Spinal Cord ◽  
Stem Cells ◽  
Glial Fibrillary Acidic Protein ◽  
Acidic Protein ◽  
Radial Glial
Sign in / Sign up

Export Citation Format

Share Document