scholarly journals Isolation and Multiple Differentiation of Rat Pericardial Fluid Cells

2021 ◽  
Vol 9 ◽  
Author(s):  
Ying Sun ◽  
Yan Wang ◽  
Zongjin Li ◽  
Zhikun Guo
Keyword(s):  
Flow Cytometry ◽  
Troponin T ◽  
Basic Research ◽  
Pericardial Fluid ◽  
Cardiac Differentiation ◽  
Good Growth ◽  
Sprague Dawley ◽  
Growth State ◽  
Pericardial Cavity ◽  
Multiple Differentiation

ObjectiveThe aim of the present study is to isolate and analyze the characterization of pericardial fluid cells (PFCs) from rat and provides a morphological basis for the basic research and clinical application of PFCs.MethodsAfter aseptic thoracotomy was performed, normal saline was injected into the pericardial cavity of 50 adult Sprague–Dawley rats. The mixture of diluted pericardial fluid was extracted, centrifuged, and cultured. The cell morphology of different generations in the pericardial fluid was observed on an inverted microscope. The expression levels of CD44, CD29, CD90, and pan-hematopoietic marker CD45 were analyzed via flow cytometry. The third-generation cells were used for osteogenic, adipogenic, and cardiac differentiation.ResultsPFCs were successfully isolated and subcultured. PFCs were predominantly circular in shape after 24 h of culture. Following subculture for 3 days, the cells demonstrated a spindle shape. The rat pericardial fluid contains cell populations with uniform morphology, good growth state, and strong proliferation ability. Flow cytometry results showed that CD29 (100%) and CD90 (99.3%) were positively expressed, whereas CD45 (0.30%) and CD44 (0.48%) were negatively expressed. The PFCs could differentiate into osteoblasts and adipocytes after being induced. Cardiac differentiation was also confirmed by cardiac troponin T (cTnT) and α-sarcomeric actin (α-SA) staining.ConclusionThis study revealed that a subpopulation of cells was isolated from pericardial fluid, which exhibited progenitor cell features and multiple differentiation potency. PFCs could serve as an alternative cell source for myocardial tissue repair, engineering, and reconstruction.

Postmortem Specificity of Troponin for Acute Miocard Infarction Diagnosis throug Qualitative Dosing from Pericardial Fluid

2018 ◽  
Vol 69 (9) ◽  
pp. 2482-2486
Author(s):  
Iuliana Hunea ◽  
Simona Irina Damian ◽  
Carmen Corina Radu ◽  
Sorin Moldoveanu ◽  
Tatiana Iov
Keyword(s):  
Sudden Cardiac Death ◽  
Cardiac Disease ◽  
Cardiac Death ◽  
Troponin I ◽  
Troponin T ◽  
Morphological Changes ◽  
Pericardial Fluid ◽  
Histological Changes ◽  
Lethal Arrhythmia ◽  
Myocardial Lesions

Cardiac disease is the leading cause of death, and sudden cardiac death occupies the first place in sudden deaths of natural causes. Sudden cardiac death due to lethal arrhythmia may be the first manifestation of a cardiac disease, such cases becoming suspect dead, thus forensic cases. The autopsy performed in such cases may reveal important cardiovascular disease but not obvious macroscopic or histological changes of acute myocardial infarction (IMA), except for cases of survival for several hours after the onset of the symptomatology. Biochemical markers were used to test for myocardial lesions in the absence of morphological changes. Methods for determining myoglobin, CK-MB, troponin T (cTn T), troponin I (cTn I) were introduced to the clinic to diagnose the condition of patients with chest pain as early as the 1990s. The lack of pathognomonic elements in corps investigations, where part of the analysis cannot be carried out, requires verification of the value of the investigations that can be carried out, with reference to the biochemical in the present case, in establishing the diagnosis with certainty.

scholarly journals A new model of chronic peripheral nerve compression for basic research and pharmaceutical drug testing

2021 ◽  
Author(s):  
Lucas Degrugillier ◽  
Katharina M Prautsch ◽  
Dirk J Schaefer ◽  
Raphael Guzman ◽  
Stefan Schären ◽  
...  
Keyword(s):  
Peripheral Nerve ◽  
Drug Testing ◽  
Basic Research ◽  
Nerve Compression ◽  
Sprague Dawley ◽  
Post Injury ◽  
New Therapeutic Approaches ◽  
Quantitative Measurements ◽  
Sprague Dawley Rats ◽  
Motor Outcomes

Aim: To develop a consistent model to standardize research in the field of chronic peripheral nerve neuropathy. Methods: The left sciatic nerve of 8-week-old Sprague–Dawley rats was compressed using a customized instrument leaving a defined post injury nerve lumen (400 μm, 250 μm, 100 μm, 0 μm) for 6 weeks. Sensory and motor outcomes were measured weekly, and histomorphology and electrophysiology after 6 weeks. Results: The findings demonstrated compression depth-dependent sensory and motor pathologies. Quantitative measurements revealed a significant myelin degeneration, axon irregularities and muscle atrophy. At the functional level, we highlighted the dynamics of the different injury profiles. Conclusion: Our novel model of chronic peripheral nerve compression is a useful tool for research on pathophysiology and new therapeutic approaches.

Abstract 855: Circulating Mesoangioblasts Differentiate Into Cardiac Myocytes And Improve Function After Acute Myocardial Infarction

Circulation ◽  
2007 ◽  
Vol 116 (suppl_16) ◽  
Author(s):  
Masayoshi Iwasaki ◽  
Masamichi Koyanagi ◽  
Stefan Rapp ◽  
Corina Schuetz ◽  
Philipp Bushoven ◽  
...  
Keyword(s):  
Myocardial Infarction ◽  
Cardiac Function ◽  
Troponin T ◽  
Marker Gene ◽  
Left Ventricular ◽  
Cardiac Differentiation ◽  
Neonatal Rat ◽  
Stem Cell Markers ◽  
Rt Pcr ◽  
Rat Cardiomyocytes

Mesoangioblasts (MAB) are vessel-associated cells identified during embryonic development. In contrast to hemangioblasts, MAB express mesenchymal (CD73) and endothelial marker, but lack the hematopoietic marker CD45. We recently identified circulating MAB in children. Children-derived MAB showed vigorous proliferation capacity and high telomerase activity. However, the potential of cardiac differentiation in these cells was not elucidated. Therefore, we tested the capacity of children-derived MAB to aquire a cardiomyogenic phenotype. MAB expressed several cardiac transcription factors such as Nkx2.5, GATA4 and MEF2C and the stem cell markers c-kit and islet-1. In order to assess cardiac differentiation capacity, we performed co-culture assays with neonatal rat cardiomyocytes (CM). Immunochemical analysis revealed that MAB expressed cardiac α-sarcomeric actinin 6 days after co-culture. Moreover, human troponin T (TnT) was expressed as demonstrated by human specific RT-PCR. To confirm these data, we examined TnT expression in MAB isolated of a 2 years old patient with a known mutation of TnT. Sequences of the cloned RT-PCR products were identical to human TnT except for the known mutation providing genetic proof of concept for cardiac differentiation. In order to exclude fusion between MAB and CM as a mechanism, we used paraformaldehyde-fixed CM as scaffold. In this assay, human TnT also was detected, indicating that differentiation is sufficient to induce cardiac marker gene expression. Next, we tested the effect of MAB to improve cardiac function. MAB were injected intramuscularly in nude mice after myocardial infarction. Functional analysis using Millar catheter 2 weeks after infarction demonstrated that cell therapy lowered filling pressure and preserved diastolic function when compared to the PBS injected group (LVEDP: −20.3%, tau: −20.6%, vs PBS injected heart). Furthermore, left ventricular volume was also decreased (LVEDV/weight −27.3%). In summary, children-derived MAB express cardiac-specific genes after co-culture with CM and improved cardiac function in vivo. Given that MAB can be easily isolated and expanded from peripheral blood, these cells might be suitable to augment cardiac repair in children with heart failure.

Abstract 20699: GMP-Grade Cardiomyocyte Differentiation Media System for Pluripotent Stem Cells in Basic and Translational Research

Circulation ◽  
2014 ◽  
Vol 130 (suppl_2) ◽  
Author(s):  
Shayne Boucher ◽  
Stacy Jones ◽  
David Kuninger ◽  
Mohan Vemuri
Keyword(s):  
Stem Cells ◽  
Flow Cytometry ◽  
Translational Research ◽  
Pluripotent Stem Cells ◽  
Troponin T ◽  
Fold Increase ◽  
Cell Number ◽  
Cardiomyocyte Differentiation ◽  
Media System ◽  
Large Numbers

Objective: Current protocols for differentiating pluripotent stem cells (PSCs) have led to heterogeneous results, varying purity levels, and long lead times for generation of cardiomyocytes. We hypothesized that a simplified and rapid cardiomyocyte differentiation media system can be developed in a scalable workflow to enable generation of large numbers of consistent, spontaneously active cardiomyocytes that could be used in basic and translational research. Methods: High quality PSCs were maintained under xenofree, feeder-free culture conditions. At time of passaging, PSC were dissociated with 0.5 mM EDTA, seeded on 1:100 Geltrex © -coated surface as small clusters at ~0.5 to 1 x 10 5 /well of a 12-well plate and maintained for four days under serum-free condition. After reaching target confluence of ~60 to 80%, an induction media was added for two days followed by addition of a second induction media for two days. After the induction step, the media was replaced with maintenance media and re-fed every other day for up to five weeks. PSC-derived cardiomyocytes were analyzed by morphology, gene expression, flow cytometry, immunocytochemistry and multi-electrode array (MEA). Results: We observed individual beating cells by Day 7 and contracting syncytia by Day 10. An over 100 fold increase in cell number was noted from the time of plating to generation of contracting syncytia of cardiomyocytes. Quantitative flow cytometry detected populations of troponin T type 2 (TNNT2)-immunoreactive cells that reached as high as 96.6%. Number of TNNT2-positive cells dropped by 20% when induced at 90% versus 60% confluency. PCR studies confirmed expression of mesoderm (T, MIXL1, MESP1), cardiac mesoderm (ISL1, GATA4, MEF2C) and mature cardiomyocyte genes (NKX2.5, TNNT2, MYH6). Immunocytochemistry studies verified expression of cardiac markers NKX2.5,GATA4, MEF2C, TNNT2 and MYH6. Initial MEA studies corroborated the presence of electrically active cells. Conclusions: We conclude that a simplified complete differentiation media system could serve as a standardized culture system for generating large numbers of consistent, spontaneously active cardiomyocytes for basic and translational research studies.

Abstract 198: Multipotent Placenta-derived Cdx2 Cells Possess in vitro and in vivo Cardiomyogenic Potential

2017 ◽  
Vol 121 (suppl_1) ◽  
Author(s):  
Sangeetha Vadakke Madathil ◽  
Amaresh Ranjan ◽  
Jesse Yoon ◽  
Joseph Tripodi ◽  
Koen Raedschelders ◽  
...  
Keyword(s):  
Stem Cell ◽  
Proteomic Analysis ◽  
Troponin T ◽  
Embryonic Stem ◽  
Late Gestation ◽  
Cardiac Differentiation ◽  
Homeodomain Protein ◽  
Mhc Molecules ◽  
Junction Protein

Stem cell-based therapies for cardiac regeneration are of crucial importance and an ideal cell-type is yet to be established. We previously reported that fetal cells from placenta “home” to injured maternal heart and approximately 40% (40/100) of the migrating cells expressed homeodomain protein Cdx2. This interesting observation led us to hypothesize that placental Cdx2 could be a novel cell target for cardiac differentiation. To understand this phenomenon, we employed a cre-lox strategy that labeled Cdx2 cells in placenta with e-GFP and induced myocardial infarction (MI) in pregnant mice at mid-gestation. The maternal heart was analyzed 4 weeks post-MI for the presence of Cdx2-eGFP-derived cardiomyocytes. Additionally, Cdx2 cells were isolated from late-gestation placenta and assayed for cardiac differentiation in vitro followed by live cell imaging. Phenotypic and whole-cell proteomic analysis, clonal and vascular lineage differentiation and immune profiling were carried out subsequently. We observed that Cdx2 cells migrated to injured maternal hearts and differentiated into cardiomyocytes highlighting the functional significance of fetal-maternal stem cell transfer. Additionally, isolated Cdx2 cells from the late placenta differentiated into spontaneously beating cardiomyocytes and expressed structural proteins cardiac troponin T(cTnT), α-sarcomeric actinin and gap junction protein Cx43. These cells underwent clonal expansion and differentiated into endothelial and smooth muscle lineages in culture indicative of their multipotent nature. Low expression of MHC molecules and other components of the immune-response, infer that these cells possess the ability to evade host immune surveillance. Proteomic analysis demonstrated that 145 proteins were uniquely identified in the Cdx2 cells compared to embryonic stem cells. These protein networks reflected an increased activation of functions involving migration, fertility, homing, and chemotaxis. Our study is the first to demonstrate that Cdx2 may play a role in cardiac differentiation and delineate multipotent cells in placenta with an inherent “homing” ability. These findings point to a potential role for Cdx2 cells in cardiac regenerative therapies using allogeneic cells.

Excretion in the Cephalopod, Octopus Dofleini

1965 ◽  
Vol 42 (1) ◽  
pp. 71-98
Author(s):  
F. M. HARRISON ◽  
A. W. MARTIN
Keyword(s):  
Glucose Concentration ◽  
Hippuric Acid ◽  
Pericardial Fluid ◽  
Active Secretion ◽  
Pericardial Cavity ◽  
Before And After ◽  
Branchial Heart ◽  
Urine Formation ◽  
Serial Samples

1. Experiments have been performed to determine some of the processes involved in urine formation in the octopus. The concentration of specific substances was followed in serial samples of the blood, pericardial fluid and urine for extended periods of time before and after the administration of metabolic poisons. 2. The results with inulin indicate that it is filtered since its concentration is approximately the same in the blood, pericardial fluid and urine. It is proposed that filtration of the blood occurs within the pericardial cavity. 3. The results with glucose indicate that it is reabsorbed from the filtrate in the pericardial cavity and reno-pericardial canal. Phlorizin administration increased the glucose concentration in the pericardial fluid and urine to the level of that in the blood. 4. The filtrate flows by way of the reno-pericardial canal into the renal sac. The results with phenolsulphonphthalein (PSP), para-amino hippuric acid (PAH) and urea indicate that active secretion into the filtrate takes place in the renal sacs. PSP, PAH and urea were in higher concentration in the urine than in the blood and pericardial fluid. The secretion of PAH and PSP was inhibited by DNP and benemid. 5. The theory of urine formation proposed is the following. Filtration of the blood occurs across the wall of the branchial heart appendage into the pericardial cavity. The filtrate formed passes by way of the reno-pericardial canal into the renal sacs. Reabsorption occurs en route to the renal sacs. Active secretion into the filtrate occurs in the renal sacs from where the filtrate is expelled to the outside as urine.

scholarly journals Acute myopericarditis caused by Salmonella enterica serovar Enteritidis: a case report

2020 ◽  
Vol 4 (4) ◽  
pp. 1-5
Author(s):  
Omar Chehab ◽  
Emma McGuire ◽  
Robert L Serafino Wani ◽  
Roshan Weerackody
Keyword(s):  
Salmonella Enterica ◽  
Troponin T ◽  
Clinical Signs ◽  
Pericardial Fluid ◽  
Salmonella Enterica Serovar Enteritidis ◽  
Salmonella Spp ◽  
Large Pericardial Effusion ◽  
New Diagnosis ◽  
Diagnosed Diabetes ◽  
Newly Diagnosed Diabetes

Abstract Background Acute myopericarditis can be caused by a myriad of infectious and non-infectious aetiologies, however, it is often considered to be due to self-limiting viral infection. Salmonella spp. myopericarditis is rare and the few cases in the literature suggest significant associated morbidity and mortality. Case summary A 44-year-old man presented with fever, dyspnoea, and chest pain. He was found to have a large pericardial effusion with clinical signs of tamponade and sepsis. Therapeutic pericardiocentesis was performed and ceftriaxone and levofloxacin were administered. Fully sensitive Salmonella enterica serovar Enteritidis (S. Enteritidis) was isolated in his pericardial fluid and he made a full recovery after a 4-week course of ciprofloxacin. A new diagnosis of type 2 diabetes mellitus was made on admission. A follow-up cardiac magnetic resonance (CMR) scan was suggestive of myocarditis which was unexpected given a normal Troponin T level on presentation. Discussion We report a rare case of S. Enteritidis myopericarditis. Our case is notable as the patient was immunocompetent apart from newly diagnosed diabetes. This case highlights the value of CMR imaging in assessing for myocarditis and ventricular function.

scholarly journals Erysipelas Complicated with Acute Exudative Pericarditis

Medicina ◽  
2020 ◽  
Vol 56 (11) ◽  
pp. 571
Author(s):  
Akvilė Gečaitė ◽  
Aušra Vainalavičiūtė ◽  
Daiva Emilija Rekienė ◽  
Laima Jankauskienė ◽  
Albinas Naudžiūnas
Keyword(s):  
Troponin I ◽  
Epigastric Pain ◽  
White Blood Cells ◽  
Pericardial Fluid ◽  
C Reactive Protein ◽  
St Segment Elevation ◽  
Pericardial Cavity ◽  
Reactive Protein ◽  
St Segment ◽  
The Face

Erysipelas is a common skin infection of the upper dermis. Its most common complications are local; these include abscess formation, skin necrosis, etc. In the present article, we introduce a case of a 75-year-old patient with erysipelas of the face complicated with acute exudative pericarditis. The patient came to Kaunas Clinical Hospital complaining of extreme fatigue and fever, oedema of the left side of the face, and erythema typical for erysipelas. The patient also felt sternum and epigastric pain, especially during breathing, and dyspnoea. Heart work was rhythmic 100 bpm; blood pressure was 142/70 mmHg. Pericardial friction rub was heard over the left sternal border. There were no alterations in other systems. In the electrocardiogram, concave ST segment elevation in leads II, III, and aVF was identified. In addition, during hospitalisation, the patient experienced atrial fibrillation paroxysm, which was treated with amiodarone intravenously. The blood test showed C-reactive protein: 286 mg/L; white blood cells: 20 × 109/L; troponin I was within the normal range. During echocardiography, pericardial fluid in pericardial cavity was identified. As no changes in troponin I were observed, according to the ST segment elevation, the woman was diagnosed with erysipelas of the left side of the face complicated with acute exudative pericarditis. Antibacterial treatment of cephalosporins was administered. After the treatment, C-reactive protein decreased to 27.8 mg/L; whereas, in the electrocardiogram, the return of the ST segment to the isoline was observed, and pericardial fluid resorbed from the pericardial cavity. To the best of the authors’ knowledge, this case is a rare combination of erysipelas complicated with acute exudative pericarditis.

398 DIFFERENTIATION OF MOUSE EMBRYONIC STEM CELLS INTO CARDIOMYOCYTES BY USING SLOW TURNING LATERAL VESSEL (STLV/BIOREACTOR)

2010 ◽  
Vol 22 (1) ◽  
pp. 355
Author(s):  
S. Rungarunlert ◽  
K. Tar ◽  
S. Muenthaisong ◽  
M. Techakumphu ◽  
M. Pirity ◽  
...  
Keyword(s):  
Large Scale ◽  
Embryoid Body ◽  
Troponin T ◽  
Es Cells ◽  
Statistical Significance ◽  
Embryonic Stem ◽  
Industrial Applications ◽  
Cardiac Differentiation ◽  
Seeding Density ◽  
Es Cell

Cardiomyocytes derived from embryonic stem (ES) cells are anticipated to be valuable for cardiovascular drug testing and disease therapies. The overall efficiency and quantity of cardiomyocytes obtained by differentiation of ES cells is still low. To enable a large-scale culture of ES-derived cells, we have tested a scalable bioprocess that allows direct embryoid body (EB) formation in a fully controlled, bioreactor/STLV (slow turning lateral vessel, Synthecon, Inc., Houston, TX, USA) following inoculation with a single cell suspension of mouse ES cells. Technical parameters for optimal cell expansion and efficient ES cell differentiation were compared, such as ES cell seeding density (3 × 105 and 5 × 105 cells mL-1) into the bioreactor and day of transfer and plating of EB on gelatinated petri dishes (Day 2, Day 3, Day 4, and Day 5). The quantity and quality of EB production including the yield and size of EB, as well as viability and apoptosis of cells, were analyzed. Furthermore, after cultivation, well-developed contracting EB with functional cardiac muscle were obtained in which the percentage of EB beating/well and several specific cardiac genes [cardiac Troponin T (cTnT) and α-actinin] expression were also determined. Data are expressed as mean ± SEM of at least 3 independent experiments. Statistical analyses included one-way ANOVA and Student’s t-test Statistical significance was set at P < 0.05. The results showed that 5 × 105 ES cells mL-1 seeded into the STLV significantly improved the homogeneity of size of EB formed compared with 3 × 105 ES cells mL-1. The EB derived from Days 2 or 3 culturing in STLV had less necrotic cells than Days 4 and 5 groups. Furthermore, plating these EB on Days 2 and 3 resulted in significantly more EB beating/well than that of Days 4 and 5 groups. For cardiac differentiation, the group with 5 × 105 ES cells mL-1 seeded into STLV and transferred and plated on Day 3 expressed more cardiac markers than other groups. In conclusion, the optimized rotary suspension culture method can produce a highly uniform population of efficiently differentiating EB in large quantities in a manner that can be easily implemented by basic research laboratories. This method provides a technological platform for the controlled large-scale generation of ES cell-derived cells for clinical and industrial applications. This work was financed by The Thailand Commission on Higher Education (CHE-PhD-SW-2005-100), EUFP6 CLONET (MRTN-CT-2006-035468), NKFP_07_1-ES2HEART-HU (OM-00202-2007), and EUFP7 (PartnErS, PIAP-GA-2008-218205).

scholarly journals P717 Isolated left ventricular tamponade in a patient with severe pulmonary hypertension due to chronic thromboembolisms: a case report

2020 ◽  
Vol 21 (Supplement_1) ◽  
Author(s):  
O Vinter ◽  
M Trbusic ◽  
M Menegoni
Keyword(s):  
Emergency Department ◽  
Pulmonary Hypertension ◽  
Pericardial Effusion ◽  
Right Ventricular ◽  
Interventricular Septum ◽  
Posterior Wall ◽  
Left Ventricular ◽  
Pericardial Fluid ◽  
Left Ventricular Cavity ◽  
Pericardial Cavity

Abstract A case presents a 37 years old patient who presented to emergency department with progression of dyspnea. Patient had a history of pulmonary hypertension due to chronic thromboembolism and known, repetitive thrombosis of both legs with both of them postthrombotically altered, especially left leg. During his emergency department workup he had a pulmonary angiography performed which showed evidence of old thromboembolism in right pulmonary main branch and circumferential pericardial effusion which was dominantly locularized behind left ventricular posterior wall. Emergency echocardiography was performed which showed marked respiratory variations in mitral and aortic flow with mid to late diastolic left ventricular collapse. Also left ventricular cavity was severely reduced ( EDD 29 mm) due to prominent interventricular septum (right ventricular pressure overload) and hyperkinetic posterior wall (pericardial effusion). There were no apparent signs of compression of right ventricular chambers. Clinically patient had no pulsus paradoxus and had an RR of 115/70 mmHg. Emergency pericardiocentesis was performed using subxiphoid approach. However, pericardiocentesis setting was challenging because patient also had ample of ascites which made orientation by aspiration impossible. Instead puncture was performed under fluoroscopy while slowly instilling the contrast until contrast was delievered intradiaphragmally. From there needle was advanced 3-4 mm into pericardial cavity and pigtail catheter was placed. A total of 2200 ml of milky pericardial fluid was removed during the following 48 hours (cytology – mixed type; triglycerides 1.9 mmol/L). Patient was initially treated with corticosteroids and colhicin, but had a relapse of pericardial effusion once drainage was stopped so re-pericardiocentesis was performed. This time a total of 7200 ml of pericardial fluid was drained so we opted for pericardial fenestration (into left pleural space). Unfortunately, patient died on the 8th postoperative day due to complications (developed subcutaneous emphysema at the place of insertion of thoracal drainage and developed respiratory, then refractory cardiac arrest).

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