47 PHYSIOLOGICAL STATUS OF MALE AND FEMALE MINIATURE PIGS CLONED WITH MESENCHYMAL STEM CELLS

2011 ◽  
Vol 23 (1) ◽  
pp. 129
Author(s):  
S. L. Lee ◽  
G. H. Maeng ◽  
W. J. Lee ◽  
R. H. Chon ◽  
G. J. Rho
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Physiological Status ◽  
Normal Range ◽  
Control Male ◽  
Control Female ◽  
Male And Female ◽  
Miniature Pigs ◽  
Complete Blood Counts ◽  
And Control

The information on the physiological health status and the endocrinological parameters of cloned pigs is limited. To address this issue, the present study evaluated the hematological, biochemical, and endocrinological status of adult cloned male and female miniature pigs. Male and female cloned miniature pigs were produced by NT using mesenchymal stem cells (MSCs) derived bone marrow of miniature pig (T-type, PWG Micro-pig®, PWG Genetics Korea, South Korea). Cloned and age-matched control male and female miniature pigs were maintained under the same conditions in a farm facility, and collected blood samples via jugular venipuncture at the age of 1 year, 3, 6, and 9 months. Complete blood counts of leukocytes, erythrocytes, and thromocytes were performed using automated hematology cell counter (MS9-5V; Melet Schloesing Lab., France). Biochemical analyses were performed using a bench-top dry chemistry analyzer (Vettest 8008 Chemistry Analyzer; IDEXX Lab., UK) by examining creatinine, glucose, blood urea nitrogen, Gamma-glutamyltransferase (γ–GGT), albumin, total bilirubin, total protein (TP), alanine aminotransferase (ALT), aspartate aminotransferase, creatine kinase, cholesterol, and amylase. Plasma growth hormone, insulin, insulin-like growth factor-1 (IGF-1), thyroid, tyroxine, cortisol, aldosterone, progesterone, testosterone, and estrogen concentration were determined by a 7020 automatic analyzer (Hitachi Ltd., Tokyo, Japan). Most parameters related to the hematological and biochemical status of cloned female and male miniature pigs were similar to control animals. However, γ–GGT (67.0 ± 20.8) and ALT (78.7 ± 24.0) levels of cloned male were higher compared to normal range (16 to 30 and 9 to 43 U L–1, respectively), and significantly (P < 0.05 and P < 0.01, respectively) higher than cloned female (GGT: 38.7 ± 2.9, ALT: 55.0 ± 16.1) and control female and male pigs (GGT: 27.5 ± 4.8 and 23.0 ± 4.4, ALT: 38.5 ± 7.9 and 32.3 ± 8.5). TP (8.2 ± 0.2) and cholesterol (87.33 ± 6.66) levels of cloned female were higher compared to normal range (6.0 to 8.0 g dL–1 and 18 to 79 mg dL–1, respectively), and significantly (P < 0.05) higher than cloned male (TP: 7.7 ± 0.4, cholesterol: 85.0 ± 8.2) and control female and male (TP: 7.9 ± 0.4 and 7.1 ± 0.6, cholesterol: 62.8 ± 3.6 and 57.0 ± 14.4). Endocrinological variation of insulin and IGF-1 of cloned female (1.43 ± 0.7 and 226.10 ± 65.0, respectively) were higher than cloned male and control female and male (0.9 ± 0.1 and 174.2 ± 42.2, 0.5 ± 0.3 and 199.9 ± 8.9, 0.5 ± 0.4 and 168.9 ± 21.2, respectively). In summary, despite similarities in hematological and biochemical parameters between cloned male and female miniature pigs and controls, a greater degree of physiological and endocrinological variations were found in cloned pigs. Based on the changes of the parameters related to growth metabolism, cloned male and female miniature pigs may have dysfunction of the liver. Therefore, the variabilities found must be taken into account before considering the cloned pigs for applications in biomedicine and xenotransplantation. This study was supported by Grant No. 2007031034040 from Bio-organ, Republic of Korea.

scholarly journals Clinical and imaging outcomes after intrathecal injection of umbilical cord tissue mesenchymal stem cells in cerebral palsy: a randomized double-blind sham-controlled clinical trial

2021 ◽  
Vol 12 (1) ◽  
Author(s):  
Man Amanat ◽  
Anahita Majmaa ◽  
Morteza Zarrabi ◽  
Masoumeh Nouri ◽  
Masood Ghahvechi Akbari ◽  
...  
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Cerebral Palsy ◽  
Umbilical Cord ◽  
Intrathecal Injection ◽  
Control Group ◽  
Umbilical Cord Tissue ◽  
The Mean ◽  
And Control ◽  
Experimental Group

Abstract Background This study assessed the safety and efficacy of intrathecal injection of umbilical cord tissue mesenchymal stem cells (UCT-MSC) in individuals with cerebral palsy (CP). The diffusion tensor imaging (DTI) was performed to evaluate the alterations in white-matter integrity. Methods Participants (4–14 years old) with spastic CP were assigned in 1:1 ratio to receive either UCT-MSC or sham procedure. Single-dose (2 × 107) cells were administered in the experimental group. Small needle pricks to the lower back were performed in the sham-control arm. All individuals were sedated to prevent awareness. The primary endpoints were the mean changes in gross motor function measure (GMFM)-66 from baseline to 12 months after procedures. The mean changes in the modified Ashworth scale (MAS), pediatric evaluation of disability inventory (PEDI), and CP quality of life (CP-QoL) were also assessed. Secondary endpoints were the mean changes in fractional anisotropy (FA) and mean diffusivity (MD) of corticospinal tract (CST) and posterior thalamic radiation (PTR). Results There were 36 participants in each group. The mean GMFM-66 scores after 12 months of intervention were significantly higher in the UCT-MSC group compared to baseline (10.65; 95%CI 5.39, 15.91) and control (β 8.07; 95%CI 1.62, 14.52; Cohen’s d 0.92). The increase was also seen in total PEDI scores (vs baseline 8.53; 95%CI 4.98, 12.08; vs control: β 6.87; 95%CI 1.52, 12.21; Cohen’s d 0.70). The mean change in MAS scores after 12 months of cell injection reduced compared to baseline (−1.0; 95%CI −1.31, −0.69) and control (β −0.72; 95%CI −1.18, −0.26; Cohen’s d 0.76). Regarding CP-QoL, mean changes in domains including friends and family, participation in activities, and communication were higher than the control group with a large effect size. The DTI analysis in the experimental group showed that mean FA increased (CST 0.032; 95%CI 0.02, 0.03. PTR 0.024; 95%CI 0.020, 0.028) and MD decreased (CST −0.035 × 10-3; 95%CI −0.04 × 10-3, −0.02 × 10-3. PTR −0.045 × 10-3; 95%CI −0.05 × 10-3, −0.03 × 10-3); compared to baseline. The mean changes were significantly higher than the control group. Conclusions The UCT-MSC transplantation was safe and may improve the clinical and imaging outcomes. Trial registration The study was registered with ClinicalTrials.gov (NCT03795974).

Abstract 2838: Bone Marrow Mesenchymal Stem Cells Differentiated into Beating Cardiomyocytes In Vivo and Improved Myocardial Function

Circulation ◽  
2007 ◽  
Vol 116 (suppl_16) ◽  
Author(s):  
Tong Wang ◽  
Wanchun Tang ◽  
Shijie Sun ◽  
Min-shan Tsai ◽  
Max Harry Weil
Keyword(s):  
Stem Cells ◽  
Bone Marrow ◽  
Mesenchymal Stem Cells ◽  
Myocardial Function ◽  
Phosphate Buffer Solution ◽  
Sprague Dawley ◽  
Buffer Solution ◽  
Marrow Mesenchymal Stem Cells ◽  
And Control

Background: In settings of heart failure, infusion of bone marrow mesenchymal stem cells (MSCs) improves myocardial function both in experimental and clinical studies. The mechanism by which MSCs improve myocardial function remains unknown. Hypothesis: MSCs may differentiate into beating myocytes in vivo. The contractility of these cells is comparable with those of myocytes. Methods: A thoracotomy was performed in 10 male Sprague-Dawley rats, weighing 350 – 450g. Myocardial infarction was induced by ligation of the left anterior descending artery (LAD). One week later, animals were randomized to receive 5×10 6 MSCs marked with PKH26 in phosphate buffer solution (PBS) or as a PBS bolus injection into local infarcted myocardium. Six weeks after the MSCs or PBS injection, the hearts were harvested and digested with collagease type II and single cardiomyocytes were obtained. PKH26 labeled myocytes differentiating from MSCs were observed with a microscope Olympus I×71. The contractility of labeled and unlabeled beating cells in MSCs-treated animals was compared. The contractility of unlabeled myocytes was compared between MSCs-treated and control groups. Result: The beating fluorescent labeled myocytes can be found in MSCs-treated animals [(1.2±0.4) ×10 6 ] and contractility of these cells were the same as that of unlabeled beating myocytes (Table 1 ). The contractility of unlabeled myocytes, however, was significantly better in MSCs-treated animals. Conclusion: MSCs could differentiate into the beating myocytes. However, this may not be the sole mechanism of improved myocardial function. Table 1 Cells contractility (%)

miR-1 Activates NF-κB to Promote the Differentiation of Bone Marrow Mesenchymal Stem Cells in Mouse Models of Glioma

2021 ◽  
Vol 11 (7) ◽  
pp. 1327-1332
Author(s):  
Long Zhou ◽  
Kui Wang ◽  
Meixia Liu ◽  
Wen Wei ◽  
Liu Liu ◽  
...  
Keyword(s):  
Stem Cells ◽  
Bone Marrow ◽  
Mesenchymal Stem Cells ◽  
Western Blot ◽  
Mouse Models ◽  
Cell Apoptosis ◽  
Bone Diseases ◽  
Control Group ◽  
Marrow Mesenchymal Stem Cells ◽  
And Control

NF-κB activation and its abnormal expression are involved in the progression of glioma. miRNA plays a crucial role in bone diseases. The role of NF-κB is becoming more and more important. The purpose of this study is to explore the mechanism by how miR-1 regulates NF-κB signaling. C57 glioma mouse models were divided into osteoporosis (OP) group and control group. qPCR was used to measure miR-1 levels in OP and control mice. Bone marrow mesenchymal stem cells (BMSCs) were cultured and transfected with miR-1 specific siRNA to establish miR-1 knockout cell model followed by analysis of cell apoptosis, expression of NF-κB signaling molecules by western blot. qPCR results showed that miR-1 levels in OP mice were significantly reduced compared to control mice. A large number of siRNA particles were observed in transfected BMSCs under a fluorescence microscope. qPCR results showed that siRNA transfection significantly suppressed miR-1, indicating successful transfection. Flow cytometry revealed significant differences in cell apoptosis between miR-1 siRNA group and the NC group. Western blot indicated miR-1 promoted BMSCs differentiation via NF-κB mediated up-regulation of ALP activity. The expression of miR-1 is low in BMSCs of mice with glioma. In addition, BMSCs differentiation is enhanced by NF-κB activation via up-regulating miR-1.

scholarly journals p130 And pRb in the Maintenance of Transient Quiescence of Mesenchymal Stem Cells

2020 ◽  
Vol 2020 ◽  
pp. 1-9
Author(s):  
Boris Popov ◽  
Nikolai Petrov ◽  
Vladimir Ryabov ◽  
Igor Evsyukov
Keyword(s):  
Cell Cycle ◽  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Critical Role ◽  
Growth Curves ◽  
State Level ◽  
Gel Shift Assay ◽  
Multipotent Mesenchymal Stem Cells ◽  
Cycle Regulation ◽  
And Control

An effective regulation of quiescence plays a key role in the differentiation, plasticity, and prevention of stem cells from becoming malignant. The state of quiescence is being controlled by the pRb family proteins which show overlapping functions in cell cycle regulation; however, their roles in controlling the proliferation of mesenchymal stem cells (MSCs) remain to be understood. This study investigated the regulation of transient quiescence using growth curves, proliferation assay, the cytometric evaluation of cell cycle, Western blotting, and the electromobility gel shift assay (EMSA) on synchronized MSCs of the C3H10Т1/2 and control cells with different statuses of pRb proteins. It has been found that functional steady-state level of p130 but not pRb plays a critical role for entering, exiting, and maintenance of transient quiescence in multipotent mesenchymal stem cells.

scholarly journals Intrasplenic Transplantation of Bioencapsulated Mesenchymal Stem Cells Improves the Recovery Rates of 90% Partial Hepatectomized Rats

2012 ◽  
Vol 2012 ◽  
pp. 1-6 ◽  
Author(s):  
Zun Chang Liu ◽  
Thomas Ming Swi Chang
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Hepatocyte Proliferation ◽  
Polymeric Membrane ◽  
Control Group ◽  
Histopathological Analysis ◽  
Recovery Rates ◽  
Sham Control Group ◽  
And Control ◽  
Intrasplenic Transplantation

Mesenchymal stem cells (MSCs) derived from bone marrow can secrete cytokines and growth factors and can transdifferentiate into liver cells. We transplanted polymeric membrane bioencapsulated MSCs into the spleens of 90% partial hepatectomized rats. This resulted in 91.6% recovery rates. This is compared to a recovery rate of 21.4% in the 90% hepatectomized rats and 25% in the 90% hepatectomized rats receiving intrasplenic transplantation of free MSCs. After 14 days, the remnant livers in the bioencapsulated MSCs group are not significantly different in weight when compared to the sham control group. From day 1 to day 3 after surgery, in the bioencapsulated MSCs group, the plasma HGF and IL-6 were significantly higher than those in the free MSCs group and control group (P<0.01); plasma TNF-αwas significantly lower (P<0.001). We concluded that the intrasplenic transplantation of bioencapsulated MSCs significantly increases the recovery rates of 90% hepatectomized rats. It is likely that the initial effect is from proliver regeneration factors followed later by the transdifferentiated hepatocyte-like cells. However, histopathological analysis and hepatocyte proliferation study will be needed to better understand the regenerative mechanisms of this result. This study has implications in improving the survival and recovery of patients with very severe liver failure due to hepatitis, trauma, or extensive surgical resection.

scholarly journals Mesenchymal stem cells modulate the gene expression of T- type Calcium Channel Subunit Alpha 1G (Cav3.1) in acute phase of epilepsy

2021 ◽  
Author(s):  
Vitoria Pimentel da Silva ◽  
Laura Provenzi ◽  
Nicole Becker ◽  
Giovani Zocche ◽  
Gabriel Leal ◽  
...  
Keyword(s):  
Gene Expression ◽  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Calcium Channel ◽  
Calcium Influx ◽  
Control Group ◽  
Administration Routes ◽  
Therapeutic Alternatives ◽  
And Control ◽  
The Brain

Introduction: Temporal Lobe Epilepsy (TLE) is a disorder caused by neuronal electrical imbalance, clinically manifested by spontaneous and recurrent seizures1,2. Its pathogenesis involves channelopathies of calcium channels, which contributes to hyperexcitability and hypersynchrony in TLE3 . About 30% of patients do not respond to drug treatment4 , making it necessary to develop new therapeutic alternatives, such as cell therapy. This work aimed to evaluate the modulation of mesenchymal stem cells (MSCs) in the calcium channel CACNA1G (Cav3.1) gene expression. Methods: MSCs were extracted from Wistar rats bone marrow and then cultured and transplanted intravenously and intranasally in the control and epileptic groups. The brain was collected 1 and 7 days after transplantation to analyze gene expression. Results: The analysis showed that treated animals had greater gene expression, compared to animals not treated in the epileptic and control group, in both days and administration routes. Furthermore, epileptic animals that were not treated had a low or negative expression of the gene. The epileptic rats that were treated, on the other hand, had a marked increase in gene expression e in the prefrontal cortex. Conclusion: This up-regulation noted on the treated groups raises the hypothesis that MSCs would be using these channels to modify the microenvironment5 , intensifying Cav.3.1 transcription and contributing to tissue regeneration by neurodifferentiation6,7. This is supported by the increase in the calcium influx present in the early stages of neuronal maturation8,9. Thus, MSCs can modulate gene expression in the pilocarpine-induced animal’s brain, making Cav3.1 a target to be explored in epilepsy.

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