Histone Demethylase KDM5B Catalyzed H3k4me3 Demethylation to Promote Differentiation of Bone Marrow Mesenchymal Stem Cells into Cardiomyocytes

Mapping Intimacies ◽

10.21203/rs.3.rs-1038705/v1 ◽

2021 ◽

Author(s):

Hongxiao li ◽

Chenlu Zhong ◽

Zhen Wang

Keyword(s):

Stem Cells ◽

Bone Marrow ◽

Mesenchymal Stem Cells ◽

Myocardial Cell ◽

Tumor Formation ◽

Control Group ◽

Pluripotency Gene ◽

Marrow Mesenchymal Stem Cells ◽

Differentiation Efficiency

Abstract BACKGROUND: Studies have shown that histone H3 methylation is involved in regulating the differentiation of Bone Marrow Mesenchymal Stem Cells (BMSCs). KDM5B can specifically reduce the level of histone 3 lysine 4 trimethylation (H3K4me3), thereby activating the expression of related genes and participating in biological processes such as cell differentiation, embryonic development and tumor formation. Whether KDM5B is involved in the regulation of BMSCs differentiation into cardiomyocytes through the above manner has not been reported.OBJECTIVE: To investigate the effect of KDM5B on the induction and differentiation of swine BMSCs into myocardial cells in vitro.METHODS: Swine bone marrow BMSCs were isolated and cultured, and the overexpression, interference expression and blank vector of KMD5B were constructed and transfected by lentivirus. BMSCs was induced to differentiate into cardiomyocytes by 5-azacytidine (5-AZA) in vitro, and the differentiation efficiency was compared by immunofluorescence, RT-PCR, Western Blot and whole-cell patch clamp detection.RESULT: Compared with the control group, the expression levels of histone H3K4me3 and pluripotency gene Nanog in KDM5B overexpression group were significantly decreased, while the expression level of key myocardial gene HCN4 was significantly increased, and the Na+ current density on the surface of differentiated myocardial cell membrane was significantly increased. Meanwhile, the corresponding results of the KDM5B silent expression group were just opposite. CONCLUSIONS: It indicated that enhanced KDM5B expression could promote the differentiation of BMSCs into cardiomyocytes and improve the differentiation efficiency by controlling H3K4 methylation levels..

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37 Healthy Foals Produced Using Bone Marrow-Mesenchymal Stem Cells as Nuclear Donors in Horse Cloning

Reproduction Fertility and Development ◽

10.1071/rdv30n1ab37 ◽

2018 ◽

Vol 30 (1) ◽

pp. 158

Author(s):

R. Olivera ◽

L. Moro ◽

R. Jordan ◽

C. Luzzani ◽

S. Miriuka ◽

...

Keyword(s):

Stem Cells ◽

Bone Marrow ◽

Mesenchymal Stem Cells ◽

Umbilical Cord ◽

Donor Cell ◽

Control Group ◽

Nuclear Reprogramming ◽

Marrow Mesenchymal Stem Cells

Somatic cell nuclear transfer efficiency is based on the capacity of the donor cell to be reset and reprogrammed to an embryonic state. So, the less differentiated the donor cells are, the more easily they could be reprogrammed by a recipient cytoplasm. Failures on appropriate nuclear reprogramming frequently lead to abnormalities associated with the placenta, umbilical cord, birthweight, and limbs. In the present study, we evaluated the efficiency of bone marrow mesenchymal stem cells (BM-MSC) compared with adult fibroblasts (AF) as nuclear donors in horse cloning and evaluated both in vitro and in vivo development of the embryos generated. Moreover, we focused on comparing the health of the foals generated and on the presence of anatomical abnormalities in foals produced from the different treatments. Embryos produced by AI, recovered by uterine flushing, and transferred to recipient mares were used as controls. All variables were analysed by Fisher test (P < 0.05). The cloning procedure was performed according to Olivera et al. (2016 PLoS One 11, e0164049, 10.1371/journal.pone.0164049). Both cleavage and blastocyst rates were higher when MSC were used as nuclear donors (P < 0.05). Cleavage rates were 85.6% (3875/4527) v. 90.2% (3095/3432) and blastocyst rates were 10.9% (492/4527) and 18.1% (622/3432) for AF and MSC groups, respectively. In the AF group, 476 blastocysts were transferred to recipient mares (232 transfers), and in the MSC group, 594 blastocysts were transferred 297 transfers). In the AI control group, 88 embryos were transferred. Pregnancies were diagnosed by transrectal ultrasonography 15 days after embryo transfer in all the groups. Pregnancy rates were similar between both cloning groups (41/232, 17.7% and 37/297, 12.5%for AF and MSC, respectively), but higher in the AI group (71/88, 80.7%). However, significant differences were observed in the birth of viable offsprings among the cloning groups. Despite similar rates of foal delivery (AF, 17/41, 41.5%; MSC, 21/37, 56.7%), a higher proportion of viable foals were obtained from the MSC group (20/37, 54.1%) compared with the AF group (9/41, 22%; P < 0.05). Surprisingly, as in the AI group (63/63, 100%), all of the viable foals obtained using MSC (20/20, 100%) were considered normal and did not show abnormalities associated with cloning. In contrast, in the AF group, only 4/9 (44.4%) were considered normal foals. The defects present in the other 5 foals were related to flexural and angular limb deformities and umbilical cord malformations. These were corrected rapidly with standard treatments or, in the case of the umbilical cords, minor surgery. This study shows for the first time that BM-MSC can be used as nuclear donors in horse cloning and that the foals obtained are as healthy as those produced by AI, showing no abnormalities related to deficiencies in nuclear reprogramming.

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Effect of NTN and Lmx1α on the Notch Signaling Pathway during the Differentiation of Human Bone Marrow Mesenchymal Stem Cells into Dopaminergic Neuron-Like Cells

Parkinson s Disease ◽

10.1155/2021/6676709 ◽

2021 ◽

Vol 2021 ◽

pp. 1-11

Author(s):

Jinhua Zhang ◽

Bo Yang ◽

Lilin Luo ◽

Linhui Li ◽

Xuantao Yang ◽

...

Keyword(s):

Stem Cells ◽

Bone Marrow ◽

Mesenchymal Stem Cells ◽

Notch Signaling ◽

Signaling Pathway ◽

Dopaminergic Neuron ◽

Notch Signaling Pathway ◽

Control Group ◽

Marrow Mesenchymal Stem Cells

Human bone marrow mesenchymal stem cells (h-BMSCs) have the potential to differentiate into dopaminergic neuron-like cells to treat Parkinson’s disease. The Notch signaling pathway has been implicated in the regulation of cell fate decisions such as differentiation of BMSCs. This study investigated changes in the expression of Notch-related genes in the differentiation of BMSCs in vitro into dopaminergic (DA) neuron-like cells. BMSCs transfected with empty lentiviral vectors served as the control group and those transfected with NTN and Lmx1α recombinant lentiviral vectors served as the experimental group. After induction and culture of NTN and Lmx1α-transfected h-BMSCs for 21 days, the cells exhibited features of dopaminergic neuron-like cells, which were observed by transmission and scanning electron microscopy and verified by immunofluorescence of tyrosine hydroxylase (TH) and dopamine transporter (DAT). These induced cells could secrete dopamine and had basic action potentials. Expression of the neural stem cell (NSC) markers, including octamer-binding protein (Oct4), paired box gene 6 (Pax6), and sex determining region Y-box 1 (SOX1), increased on day 14 of induction and decreased on day 21 of induction during differentiation. The human Notch signaling pathway PCR array showed a differential expression of Notch-related genes during the differentiation of h-BMSCs into DA neuron-like cells in vitro relative to that in the control group. In conclusion, h-BMSCs overexpressing NTN and Lmx1α can successfully be induced to differentiate into dopaminergic neuron-like cells with a neuronal phenotype exhibiting fundamental biological functions in vitro, and NTN and Lmx1α may affect the expression of Notch-related genes during differentiation.

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Effects of bone marrow mesenchymal stem cell-derived exosomes on bone formation on titanium

10.21203/rs.3.rs-84302/v1 ◽

2020 ◽

Author(s):

Xiaoling Zhang ◽

Liangzhi Du ◽

Ningbo Zhao ◽

Lizhe Zhu ◽

Lei Wang ◽

...

Keyword(s):

Stem Cells ◽

Bone Marrow ◽

Mesenchymal Stem Cells ◽

Bone Formation ◽

Type I Collagen ◽

Control Group ◽

Type I ◽

Oral Implants ◽

Marrow Mesenchymal Stem Cells

Abstract Background: In recent years, researchers have found that exosomes, an important component of intercellular signal transduction and exchange, have great significance in bone tissue repair. In this study, to further promote the development of oral implants, preliminary in vitro experiments were conducted to verify the different concentrations of exosomes from bone marrow mesenchymal stem cells (BMSC-exos) for osteogenesis on the surfaces of titanium sheets.Methods: In this experiment, rabbit bone marrow mesenchymal stem cells(BMSCs) were seeded on the surfaces of 10 mm × 10 mm × 1 mm square titanium sheets and were divided into four groups to investigate their adsorption, proliferation and osteogenesis after treatment with different concentrations of BMSC-exos: 1. BMSCs + titanium + 0 µg/ml BMSC-exos; 2. BMSCs + titanium + 10 µg/ml BMSC-exos; 3. BMSCs + titanium + 25 µg/ml BMSC-exos; and 4. BMSCs + titanium + 50 µg/ml BMSC-exos.Results: Compared with the control group, BMSCs’ adsorption, extension, proliferation and osteogenesis on titanium sheets were significantly increased in the Exosomes group.Conclusions: Exosomes can promote the bone formation of BMSCs on titanium plates by promoting adsorption, extension, proliferation, production of alkaline phosphatase(ALP) and type I collagen and mineralization during the osteogenesis process.

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