Electrical superior vena cava isolation using photodynamic therapy in a canine model.

AIMS: With the new era of multi-tip radiofrequency or balloon ablation catheters replacing the point-to-point ablation strategy, we aimed to determine the feasibility of a ring-laser catheter ablation technology to electrically isolate the superior vena cava (SVC) by exploring the advantages of the limitless catheter tip size possibly with the photodynamic therapy (PDT)-mediated ablation. METHODS AND RESULTS: We developed a first-generation prototype of a circular-laser-mapping catheter by fitting a 7 cm plastic optical fibre onto a circular variable-loop Lasso™ mapping catheter. Following SVC venography, both the laser catheter and another ring catheter for monitoring the SVC potentials were placed at the SVC. After the systemic infusion of a photosensitizer (talaporfin sodium), we initiated the irradiation with an output of 1 W in three canines and 0.3 W in four. The creation of electrical isolation as well as occurrence of phrenic nerve injury, sinus node injury, and SVC stenosis were evaluated before, immediately after, and 1 month after the procedure. A PDT-mediated SVC isolation was successfully performed in all seven canines. The isolation was completed with a laser irradiation of 70.4 ± 71.4 J/cm under 30.9 ± 5.0 µg/mL of a photosensitizer without any sinus node injury, phrenic nerve palsy, or SVC stenosis in both the acute and chronic evaluations. The minimum isolation time of 270 s was not correlated with the laser input power or the photosensitizer concentration. CONCLUSION: The electrical SVC isolation was successfully and instantly achieved using the PDT laser-ring catheter without any complications.

Optimal conditions for cardiac catheter ablation using photodynamic therapy.

AIMS: Photodynamic therapy (PDT) is based on non-thermal injury mediated by singlet oxygen species and is used clinically in cancer therapy. In our continuing efforts to apply this technology to cardiac catheter ablation, we clarified the optimal condition for creating PDT-mediated lesions using a laser catheter. METHODS AND RESULTS: In a total of 35 canines, we applied a laser directly to the epicardium of the beating heart during open-chest surgery at 15 min after administration of a photosensitizer, talaporfin sodium. We evaluated the lesion size (depth and width) using hematoxylin-eosin staining under varying conditions as follows: laser output (5, 10, 20 W/cm(2)), irradiation time (0-60 s), photosensitizer concentration (0, 2.5, 5 mg/kg), blood oxygen concentration (103.5 ± 2.1 vs. 548.0 ± 18.4 torr), and contact force applied during irradiations (low: <20 g, high: >20 g). A laser irradiation at 20 W/cm(2) for 60 s under 5 mg/kg (29 µg/mL) of photosensitizer induced a lesion 8.7 ± 0.8 mm deep and 5.2 ± 0.2 mm wide. The lesion size was thus positively correlated to the laser power, irradiation time, and photosensitizer concentration, and was independent of the applied contact force and oxygen concentration. In addition, the concentration of the photosensitizer strongly correlated with the changes in the pulse oximetry data and fluorescence of the backscattering laser, suggesting that a clinically appropriate condition could be estimated in real time. CONCLUSION: Photodynamic therapy-mediated cardiac lesions might be controllable by regulating the photosensitizer concentration, laser output, and irradiation time.

Electrophysiological properties of the superior vena cava and venoatrial junction in patients with atrial fibrillation: relevance to catheter ablation.

BACKGROUND: Although the superior vena cava (SVC) has been well known to be one of the important foci triggering atrial fibrillation (AF), its electrophysiological characteristics have received little research attention. The aim of this study was to investigate the electrophysiological properties of the SVC and venoatrial junction (VAJ). METHODS: Twenty-five consecutive AF patients without structural heart disease undergoing electrical SVC isolation were included in this study. After pulmonary vein isolation, a circular decapolar catheter and 2 multipolar catheters were emplaced in the VAJ, right atrial appendage (RAA), and SVC, respectively. Burst pacing and single extrastimulus were applied from the RAA and SVC. The atrial and caval potentials on the circular catheter in the VAJ were investigated. RESULTS: Intracaval conduction delay and various degrees of conduction block over the VAJ were observed with burst pacing from both the RAA and SVC. A single extrastimulus from the RAA and SVC with a basic cycle length of 600 milliseconds prolonged the conduction time via the VAJ by 81 ± 49.7 milliseconds and 61 ± 58.7 milliseconds, respectively. The atrial and caval electrograms at the VAJ, which were separated from each other by pacing applications, facilitated mapping of the earliest activation site at the VAJ. CONCLUSIONS: Intracaval conduction delay and decremental conduction property via the VAJ were demonstrated using pacing maneuvers. Pacing applications from the RAA or SVC can help distinguish the atrial and caval potentials and can facilitate mapping of the optimal ablation sites to isolate the SVC.

Effects of the selective KACh channel blocker NTC-801 on atrial fibrillation in a canine model of atrial tachypacing: comparison with class Ic and III drugs.

The present study examines the effects of NTC-801, a highly selective acetylcholine (ACh) receptor-activated potassium (KACh) channel blocker, on atrial fibrillation (AF) in a canine model with electrical remodeling. An experimental substrate for AF was created in dogs via left atrial (LA) tachypacing (400 bpm, 3-5 weeks). NTC-801, dofetilide, and flecainide were intravenously infused for 15 minutes, and the effects on AF inducibility, atrial effective refractory period (ERP), and atrial conduction velocity were examined. The effect of NTC-801 on AF termination was also evaluated. Atrial ERP was shortened and AF inducibility was increased after LA tachypacing. NTC-801 (0.3-3 µg·kg⁻¹·min⁻¹) prolonged atrial ERP irrespective of stimulation frequency and dose-dependently decreased AF inducibility. Dofetilide (5.3 µg·kg⁻¹·min⁻¹) and flecainide (0.13 mg·kg⁻¹·min⁻¹) did not significantly inhibit AF inducibility and minimally affected atrial ERP. Flecainide decreased atrial conduction velocity, whereas NTC-801 and dofetilide did not. NTC-801 (0.1 mg/kg) converted AF to normal sinus rhythm. In summary, NTC-801 exerted more effective antiarrhythmic effects than dofetilide and flecainide in a canine LA-tachypacing AF model. The antiarrhythmic activity of NTC-801 was probably due to prolonging atrial ERP independently of stimulation frequency. These results suggest that NTC-801 could prevent AF more effectively in the setting of atrial electrical remodeling.

Sema3a maintains normal heart rhythm through sympathetic innervation patterning.

Sympathetic innervation is critical for effective cardiac function. However, the developmental and regulatory mechanisms determining the density and patterning of cardiac sympathetic innervation remain unclear, as does the role of this innervation in arrhythmogenesis. Here we show that a neural chemorepellent, Sema3a, establishes cardiac sympathetic innervation patterning. Sema3a is abundantly expressed in the trabecular layer in early-stage embryos but is restricted to Purkinje fibers after birth, forming an epicardial-to-endocardial transmural sympathetic innervation patterning. Sema3a(-/-) mice lacked a cardiac sympathetic innervation gradient and exhibited stellate ganglia malformation, which led to marked sinus bradycardia due to sympathetic dysfunction. Cardiac-specific overexpression of Sema3a in transgenic mice (SemaTG) was associated with reduced sympathetic innervation and attenuation of the epicardial-to-endocardial innervation gradient. SemaTG mice demonstrated sudden death and susceptibility to ventricular tachycardia, due to catecholamine supersensitivity and prolongation of the action potential duration. We conclude that appropriate cardiac Sema3a expression is needed for sympathetic innervation patterning and is critical for heart rate control.

Idiopathic ventricular tachycardia cured by radiofrequency application from the distal great cardiac vein and the left coronary cusp.

A 79 year-old male without structural heart disease suffered from drug refractory ventricular tachycardia (VT). VTs and premature ventricular complexes (PVCs) with the same morphology occurred incessantly with a concordant R pattern in chest leads and a tall R in Lead II, III, and aVF. The origin was expected to be near the left epicardial ventricular outflow tract (LVOT), which was termed the left ventricular summit area. Pace-mapping from the LVOT and the left coronary cusp (LCC) did not match well with the QRS morphology of the PVC. A good match was obtained from the distal great cardiac vein (GCV), and radiofrequency (RF) delivery eliminated the PVC and VT. However, the PVC recurred four times upon cessation of RF delivery. By placing an ablation catheter at the LCC, we obtained pace-mapping showing two different types of QRS morphologies; one was an rS pattern in V1, and the other was an R pattern in V1 with a longer stimulus to QRS interval, which was a nearly perfect match to the PVC. RF application to the LCC permanently eliminated PVCs and VTs. Several VTs from the epicardial LVOT can be cured by RF application from both the distal GCV and the LCC.

Dystrophin conferral using human endothelium expressing HLA-E in the non-immunosuppressive murine model of Duchenne muscular dystrophy.

Human leukocyte antigen (HLA)-E is a non-classical major histocompatibility complex class I (Ib) molecule, which plays an important role in immunosuppression. In this study, we investigated the immunomodulating effect of HLA-E in a xenogeneic system, using human placental artery-derived endothelial (hPAE) cells expressing HLA-E in a mouse model. In vitro cell lysis analysis by primed lymphocytes in combination with siRNA transfection showed that HLA-E is necessary for inhibition of the immune response. Similarly, in vivo cell implantation analysis with siRNA-mediated down-regulation of HLA-E demonstrates that HLA-E is involved in immunosuppression. As hPAE cells efficiently transdifferentiate into myoblasts/myocytes in vitro, we transplanted the cells into mdx mice, a model of Duchenne muscular dystrophy. hPAE cells conferred dystrophin to myocytes of the 'immunocompetent' mdx mice with extremely high efficiency. These findings suggest that HLA-E-expressing cells with a myogenic potential represent a promising source for cell-based therapy of patients with muscular dystrophy.

Autoimmunity against M2muscarinic acetylcholine receptor induces myocarditis and leads to a dilated cardiomyopathy-like phenotype.

Patients with dilated cardiomyopathy (DCM) often have autoantibodies against cardiac antigens including the M(2) muscarinic acetylcholine receptor (M(2)R). To elucidate the role of autoimmunity against M(2)R in disease development, we induced an immune response against M(2)R by adoptive transfer into Rag2(-/-) mice of splenocytes from M(2)R(-/-) mice immunized with a recombinant M(2)R protein. T lymphocytes transiently infiltrated the heart in recipient mice followed by morphological changes in cardiomyocytes. These mice produced IgG antibodies against M(2)R, which bound to cardiomyocytes in vivo and decreased the amplitude of calcium signals in isolated rat cardiomyocytes in vitro. Recipient mice showed increased heart weights associated with increased intraventricular diameter, decreased systolic function, and increased action potential duration, which are characteristics of DCM. Our results suggest that myocarditis and DCM associated with the presence of anti-M(2)R antibodies are autoimmune diseases with a risk of progressing to the terminal stage. Our mouse model will be useful in the analysis of the molecular mechanisms of disease progression and the development of new therapies for DCM.

Ridge-related reentry: a variant of perimitral atrial tachycardia.

INTRODUCTION: The ridge between the left pulmonary veins (PV) and the left atrial appendage composes part of the lateral mitral isthmus (LMI). Following circumferential PV isolation and LMI linear ablation for the treatment of atrial fibrillation (AF), a critical pathway might develop over the ridge leading to a ridge-related reentry (RRR). METHODS AND RESULTS: Out of 61 patients who underwent circumferential PV isolation appended by LMI ablation, 5 patients developed RRR. The diagnosis of RRR was based on (1) macro-reentrant atrial tachycardia involving the septum, anterior and inferior wall of the left atrium; (2) slow conduction along the ridge; (3) wide-split double potentials in the ventricular aspect of the LMI were identified with the coronary sinus (CS) electrodes. RRR was investigated with electroanatomical mapping and entrainment mapping and catheter ablation was carried out in all patients. The mean cycle length (CL) of RRR was 312 ± 82 milliseconds and the PPIs at the left atrial septum, inferior and anterior wall during RRR were 10 ± 6, 12 ± 8, 9 ± 5 milliseconds longer than the RRR CL. The interval of the double potentials recorded in the CS electrodes crossing the LMI was 164 ± 38 milliseconds during RRR and the PPI on the LMI near the mitral annulus was 57 ± 10 milliseconds longer than the RRR CL. Catheter ablation was performed anatomically by targeting the ridge and successfully terminated RRR. CONCLUSION: After circumferential PV isolation and ablation for LMI in patients with AF, RRR can develop by utilizing the surviving myocardial tissue of the ridge as a critical pathway.

Three-dimensional imaging and mapping of the right and left phrenic nerves: relevance to interventional cardiovascular therapy.

AIMS: Phrenic nerves (PNs) can be damaged during interventional cardiovascular therapy because of the nerves' proximity to the heart. This study aimed to analyse the anatomy of the PN by performing three-dimensional (3-D) imaging and pace mapping. METHODS AND RESULTS: Forty consecutive patients with atrial fibrillation referred for catheter ablation were enrolled in this study and underwent preoperative cardiovascular computed tomography (CT). In 10 patients with sinus rhythm during tomography, 3-D images of the right and left pericardiophrenic bundles (PBs), consisting of the ipsilateral PN and accompanying vessels, were reconstructed from the CT data. During the electrophysiological study, PN pace mapping was performed from both atria. The course of the PBs generated by CT imaging and the PN pace map generated by the 3-D mapping system were compared. By electrical pacing, the PNs were captured in 40 individuals (100%) from the superior vena cava and the right atrium, and in 17 patients (43%) from the left atrial appendage. Clear 3-D images of PBs were reconstructed in all cases in which CT-reconstruction was performed. The distance between the locations of the right PB generated by CT imaging and those of the right PN-capture sites in the right-sided heart on the mapping system was 8.7 ± 5.8 mm. CONCLUSIONS: The 3-D routes of the bilateral PNs passing near the heart were verified by pace mapping. The preoperatively reconstructed 3-D course of the PB succeeded in locating the PN, which may facilitate the comprehension of PN anatomy to avoid its injury during interventional cardiovascular therapy.

Pretreatment of human mesenchymal stem cells with pioglitazone improved efficiency of cardiomyogenic transdifferentiation and cardiac function.

The efficacy of transplantation of default human marrow-derived mesenchymal stem cells (MSCs) was modest. In this study, our challenge was to improve the efficacy of MSC transplantation in vivo by pretreatment of MSCs with pioglitazone. MSCs were cultured with or without medium containing 1 µM of pioglitazone before cardiomyogenic induction. After cardiomyogenic induction in vitro, cardiomyogenic transdifferentiation efficiency (CTE) was calculated by immunocytochemistry using anti-cardiac troponin-I antibody. For the in vivo experiments, myocardial infarction (MI) at the anterior left ventricle was made in nude rats. Two weeks after MI, MSCs pretreated with pioglitazone (p-BM; n = 30) or without pioglitazone (BM; n = 17) were injected, and then survived for 2 weeks. We compared left ventricular function by echocardiogram and immunohistochemistry to observe cardiomyogenic transdifferentiation in vivo. Pretreatment with pioglitazone significantly increased the CTE in vitro (1.9% ± 0.2% n = 47 vs. 39.5% ± 4.7% n = 13, p < .05). Transplantation of pioglitazone pretreated MSCs significantly improved change in left ventricular % fractional shortening (BM; -4.8% ± 2.1%, vs. p-BM; 5.2% ± 1.5%). Immunohistochemistry revealed significant improvement of cardiomyogenic transdifferentiation in p-BM in vivo (BM; 0% ± 0% n = 5, vs. p-BM; 0.077% ± 0.041% n = 5). Transplantation of pioglitazone-pretreated MSCs significantly improved cardiac function and can be a promising cardiac stem cell source to expect cardiomyogenesis.

Treatment of human mesenchymal stem cells with angiotensin receptor blocker improved efficiency of cardiomyogenic transdifferentiation and improved cardiac function via angiogenesis.

To improve the modest efficacy of mesenchymal stem cell (MSC) transplantation, the treatment of human MSCs with angiotensin receptor blockers (ARBs) was investigated. MSCs were cultured with or without the medium containing 3 µmol/l of ARBs before cardiomyogenic induction. After cardiomyogenic induction in vitro, cardiomyogenic transdifferentiation efficiency (CTE) was calculated by immunocytochemistry using anticardiac troponin-I antibody. In the nude rat chronic myocardial infarction model, we injected MSCs pretreated with candesartan (A-BM; n = 18) or injected MSCs without pretreatment of candesartan (BM; n = 25), each having survived for 2 weeks. The left ventricular function, as measured by echocardiogram, was compared with cardiomyogenic transdifferentiation in vivo, as determined by immunohistochemistry. Pretreatment with ARBs significantly increased the CTE in vitro (10.1 ± 0.8 n = 12 vs. 4.6 ± 0.3% n = 25, p < .05). Transplantation of candesartan-pretreated MSCs significantly improved the change in left ventricular ejection fraction (BM; -7.2 ± 2.0 vs. A-BM; 3.3 ± 2.3%). Immunohistochemistry revealed significant improvement of cardiomyogenic transdifferentiation in A-BM in vivo (BM; 0 ± 0 vs. A-BM; 0.014 ± 0.006%). Transplantation of ARB-pretreated MSCs significantly improved cardiac function and can be a promising cardiac stem cell source from which to expect cardiomyogenesis.

Xenografted human amniotic membrane-derived mesenchymal stem cells are immunologically tolerated and transdifferentiated into cardiomyocytes.

RATIONALE: Amniotic membrane is known to have the ability to transdifferentiate into multiple organs and is expected to stimulate a reduced immunologic reaction. OBJECTIVE: Determine whether human amniotic membrane-derived mesenchymal cells (hAMCs) can be an ideal allograftable stem cell source for cardiac regenerative medicine. METHODS AND RESULTS: We established hAMCs. After cardiomyogenic induction in vitro, hAMCs beat spontaneously, and the calculated cardiomyogenic transdifferentiation efficiency was 33%. Transplantation of hAMCs 2 weeks after myocardial infarction improved impaired left ventricular fractional shortening measured by echocardiogram (34+/-2% [n=8] to 39+/-2% [n=11]; P<0.05) and decreased myocardial fibrosis area (18+/-1% [n=9] to 13+/-1% [n=10]; P<0.05), significantly. Furthermore hAMCs transplanted into the infarcted myocardium of Wistar rats were transdifferentiated into cardiomyocytes in situ and survived for more than 4 weeks after the transplantation without using any immunosuppressant. Immunologic tolerance was caused by the hAMC-derived HLA-G expression, lack of MHC expression of hAMCs, and activation of FOXP3-positive regulatory T cells. Administration of IL-10 or progesterone, which is known to play an important role in feto-maternal tolerance during pregnancy, markedly increased HLA-G expression in hAMCs in vitro and, surprisingly, also increased cardiomyogenic transdifferentiation efficiency in vitro and in vivo. CONCLUSIONS: Because hAMCs have a high ability to transdifferentiate into cardiomyocytes and to acquire immunologic tolerance in vivo, they can be a promising cellular source for allograftable stem cells for cardiac regenerative medicine.

Electrical isolation of the superior vena cava using upstream phrenic pacing to avoid phrenic nerve injury.

BACKGROUND: Phrenic nerve (PN) injury is a potential complication that can occur during superior vena cava (SVC) isolation to cure atrial fibrillation (AF). Avoiding radiofrequency (RF) energy delivery is the safer alternative but may result in failed isolation. High-output PN pacing above the ablation site (upstream PN pacing) to confirm whether the PN is intact is a promising technique to avoid PN injury. This study was conducted to elucidate the safety of delivering RF energy at the site of capture of the right PN using upstream high-output pacing during electrical SVC isolation. METHODS: SVC isolation was conducted in 41 drug-resistant AF patients. When high-output pacing (25 mA) from the distal tip of the ablation catheter captured the PN at the right atrial-SVC junction, upstream PN pacing (cycle length: 1000-1500 ms) was applied during RF delivery. The application of RF energy was stopped upon the failure or weakness of diaphragmatic twitching. The feasibility of SVC isolation using upstream PN pacing was investigated. RESULTS: In all 41 patients, SVC isolation was successfully achieved. RF energy was delivered at the PN capture site in 26 patients (154 ± 138 second, 18 ± 5 W), and upstream PN pacing was successfully applied in all of the patients. Out of 46 SVC isolations, including five repeated sessions, PN injury occurred in one patient, who recovered spontaneously within 2 weeks. CONCLUSIONS: Upstream PN pacing may be effective for the safe completion of SVC isolation and to reduce the severity of PN injury.

Study of blood charring precursor states using backscattering at 663 nm from blood and optical window boundary.

BACKGROUND AND OBJECTIVE: Contact laser irradiation is generally used in therapeutic laser procedures such as plastic surgery and laser catheter lead removal. However, it may induce blood charring on the surface of the optical window in blood circumstance so that the laser beam might be blocked. Various charring detection methods have been proposed, but they detect charring only after charring has occurred. This study investigates the transient behavior of red blood cells (RBCs) prior to the charring on the surface of an optical window during red laser irradiation in blood circumstance. MATERIALS AND METHODS: The backscattering light power was continuously measured to investigate the transient behavior of a 1-mm-thick porcine blood model (hematocrit: 40%) during continuous laser irradiation (center wavelength: 663 nm; irradiance: 81 W/cm(2)). A rabbit blood model was microscopically observed after irradiation. The absorption coefficient (µ(a)) and the reduced scattering coefficient (µ'(s)) were measured using a double integrating sphere setup and the inverse adding-doubling method. The backscattering light power was continuously measured in vivo during contact laser irradiation via a laser catheter in a porcine heart cavity. RESULTS: The results reveal that it may be possible to detect a precursory state of charring from a time course of the backscattering light power. µ(a) increased monotonically by 15% until charring occurred. µ'(s) decreased by 10% followed a broad peak until charring occurred. These changes in the optical property correspond to changes in the morphology of RBCs. Changes in the backscattering light power measured in vivo were similar to those measured ex vivo. CONCLUSIONS: The transient optical changes in blood prior to charring may be caused by changes in the morphology of RBCs on the optical window surface. Backscattering light power measurements may be a practical method to detect the precursor state of charring.

Myocardial electrical conduction block induced by photosensitization reaction in exposed porcine hearts in vivo.

BACKGROUND AND OBJECTIVE: This study proposes photosensitization reaction for non-thermal cardiac ablation in arrhythmia therapy. Acute and chronic phase experiments were conducted in exposed porcine hearts to demonstrate the photosensitization reaction-induced myocardial electrical conduction block in vivo. STUDY DESIGN/MATERIALS AND METHODS: The porcine left atrial appendage was exposed under an open-chest procedure. Then, a water-soluble chlorin photosensitizer, NPe6, was injected into the pigs intravenously at 5 or 10 mg/kg. About 15 or 30 minutes after the injection, a 663-nm continuous-wave diode laser was irradiated on the surface of the atrial appendage through a silica optical fiber. The laser energy was delivered to the tissue point by point at an energy density of 50-208 J/cm(2). RESULTS: Acute and chronic tissue damages as a result of the photosensitization reaction were determined by electrophysiology and histology, respectively. The change in the myocardial conduction time between two electrodes was measured immediately after the completion of the 35-mm irradiation line between the electrodes. The conduction delay of 35.5 milliseconds might be due to the change in the conduction pathway induced by transmural acute conduction block with the photosensitization reaction. The tissue temperature increase in the irradiated area was approximately 12.8°C. Azan-staining revealed about 1-mm transmural fibrosis of the atrial appendage at 2 weeks after the irradiation (50 J/cm(2)). CONCLUSIONS: The results suggest that the photosensitization reaction might induce acute and chronic myocardial electrical conduction block. Cardiac ablation with the photosensitization reaction might be a non-temperature-mediated methodology for arrhythmia therapy.

Serum-independent cardiomyogenic transdifferentiation in human endometrium-derived mesenchymal cells.

Media with high concentrations of serum are commonly used to induce cardiomyogenic transdifferentiation in mesenchymal stem cells; however, serum contains numerous unknown growth factors and interferes with definition of specific cardiomyogenic transdifferentiation factors secreted from feeder cells. In the present study, we determined whether the transdifferentiation of human mesenchymal cells can be observed in a FBS-free medium. The efficiency of transdifferentiation was observed in 10% FBS-containing standard medium (10%FBS) and in FBS-free medium containing insulin and thyroxin (FBS-free). In the present study, we used human uterine endometrium-derived mesenchymal cells (EMC100, EMC214) and menstrual blood-derived mesenchymal cells (MMCs). After cardiomyogenic transdifferentiation, the efficiency and physiological properties of cardiomyogenesis (fractional shortening of the cell [%FS] and action potential [AP]) were evaluated. The efficiency of transdifferentiation in EMC100 and in MMCs increased 36%* and 163%* (*P < 0.05), respectively. The %FS in EMCs increased to 103%*. AP-duration more than 250 ms with a marked plateau was only observed in FBS-free (3/19), and not in 10% FBS (0/41). The cardiomyogenic transdifferentiation of human mesenchymal cells can be observed in the FBS-free medium. Phenotypes of generated cardiomyocytes were significantly more physiological in FBS-free than in 10% FBS.

Novel cardiac precursor-like cells from human menstrual blood-derived mesenchymal cells.

Stem cell therapy can help repair damaged heart tissue. Yet many of the suitable cells currently identified for human use are difficult to obtain and involve invasive procedures. In our search for novel stem cells with a higher cardiomyogenic potential than those available from bone marrow, we discovered that potent cardiac precursor-like cells can be harvested from human menstrual blood. This represents a new, noninvasive, and potent source of cardiac stem cell therapeutic material. We demonstrate that menstrual blood-derived mesenchymal cells (MMCs) began beating spontaneously after induction, exhibiting cardiomyocyte-specific action potentials. Cardiac troponin-I-positive cardiomyocytes accounted for 27%-32% of the MMCs in vitro. The MMCs proliferated, on average, 28 generations without affecting cardiomyogenic transdifferentiation ability, and expressed mRNA of GATA-4 before cardiomyogenic induction. Hypothesizing that the majority of cardiomyogenic cells in MMCs originated from detached uterine endometrial glands, we established monoclonal endometrial gland-derived mesenchymal cells (EMCs), 76%-97% of which transdifferentiated into cardiac cells in vitro. Both EMCs and MMCs were positive for CD29, CD105 and negative for CD34, CD45. EMCs engrafted onto a recipient's heart using a novel 3-dimensional EMC cell sheet manipulation transdifferentiated into cardiac tissue layer in vivo. Transplanted MMCs also significantly restored impaired cardiac function, decreasing the myocardial infarction (MI) area in the nude rat model, with tissue of MMC-derived cardiomyocytes observed in the MI area in vivo. Thus, MMCs appear to be a potential novel, easily accessible source of material for cardiac stem cell-based therapy.

Pulsatile cardiac tissue grafts using a novel three-dimensional cell sheet manipulation technique functionally integrates with the host heart, in vivo.

We devised a method of fabricating easily transplantable scaffoldless 3D heart tissue, made with a novel cell-sheet (CS) technology from cultured cardiomyocytes using a fibrin polymer coated dish. In the present study, we tested in vivo electrical communication which is essential for improving heart function between the host heart and the grafted CS. The epicardial surface of the ventricle of an anesthetized open-chest nude rat was ablated by applying a heated metal. Bilayered CS was obtained from neonatal rat primary culture. CS was transplanted onto the injured myocardial surface (sMI) (sMI+sheet group). The rats were allowed to recover for 1 to 4 weeks, to stabilize the grafts. Action potentials (APs) from the excised perfused heart were monitored by the fluorescence signal of di-4ANEPPS with a high speed charge-coupled device camera. The APs were observed under epicardial pacing of the host heart or the CS grafts. The pacing threshold of the current output was measured in the sMI+sheet group and in the nongrafted sMI group at the center of the sMI and in the normal zone (Nz). Bidirectional AP propagation between the sMI and Nz was observed in the sMI+sheet group (n=14), but was blocked at the marginal area of the sMI in the sMI group (n=9). The ratio of the pacing threshold (sMI/Nz) was significantly lower in the sMI+sheet than in the sMI group (3.0+/-0.7, 19.0+/-6.1 respectively P<0.05). There were neither spontaneous nor pacing-induced arrhythmias in these two groups. Bidirectional smooth AP propagation between the host heart and the grafted CS was observed. This finding suggested functional integration of this CS graft with the host heart without serious arrhythmia.

Cardiac cell therapy and arrhythmias.

Cardiac stem cell based therapy is a promising therapy for patients with severe heart failure. Many types of stem cells, such as embryonic stem cells, myoblasts, marrow-derived mesenchymal stem cells, circulating endothelial progenitor cells, and cardiac precursor cells etc, are known as cellular sources for cardiac stem cell therapy. Both in the clinical and experimental setting, stem cells are reported, and supposed, to cause some arrhythmogenic adverse effects. In order to overcome these serious adverse effects, it is necessary to know the electrophysiological properties of stem cell-derived cardiomyocytes, and have a profound insight into the mechanisms of arrhythmia to know whether such arrhythmogenic properties of the cells can cause serious arrhythmia in situ. In the present study, recent publications that focus on the electrophysiological aspect of stem cell based therapy are reviewed and, furthermore, a new perspective on cardiac stem cell therapy of arrhythmias is given.