4 nya forskningsrapporter

8 hjärtforskare från San Francisco och San Jose har studerat nyfödda däggdjurs hjärtceller och deras utveckling. Man konstaterar att förmågan till nybildande (regenering) av hjärtceller avtar efter första veckan.Denna nya kunskap kommer ha bäring på framtida Reg Med forskning beträffande regenering av hjärtceller hos patienter med hjärtrelaterade skador som ex infarkt där hjärtats muskelceller skadats eller gått förlorade. För att nå dessa nya insikter har man använt sig av HoloMonitor samt fått assistans av PHI`s chefsforskare Kersti Alm.

Quantitative label-free digital holographic imaging of cardiomyocyte optical volume, nucleation, and cell division

Abstract

Cardiac regeneration in newborn rodents depends on the ability of pre-existing cardiomyocytes to proliferate and divide. This capacity is lost within the first week of postnatal development when these cells rapidly switch from hyperplasia to hypertrophy, withdraw from the cell cycle, become binucleated, and increase in size. How these dynamic changes in cell size and nucleation impact cardiomyocyte proliferative potential is not well understood. In this study, we innovate the application of a commercially available digital holographic imaging microscope, the Holomonitor M4, to evaluate the proliferative responses of mononucleated and binucleated cardiomyocytes after CHIR99021 treatment, a model proliferative stimulus. This system enables long-term label-free quantitative tracking of primary cardiomyocyte dynamics in real-time with single-cell resolution. Our results confirm that chemical inhibition of glycogen synthase kinase 3 with CHIR99021 promotes complete cell division of both mononucleated and binucleated cardiomyocytes with high frequency. Quantitative tracking of cardiomyocyte volume dynamics during these proliferative events revealed that both mononucleated and binucleated cardiomyocytes reach a similar size-increase threshold prior to attempted cell division. Binucleated cardiomyocytes attempt to divide with lower frequency than mononucleated cardiomyocytes, which may be associated with inadequate increases in cell size. By defining the interrelationship between cardiomyocyte size, nucleation, and cell cycle control, we may better understand the cellular mechanisms that drive the loss of mammalian cardiac regenerative capacity after birth.

In this study, we innovate the application of a commercially available digital holographic phase imaging microscope, the HoloMonitor M4 (Phase Holographic Imaging; Lund, Sweden), to investigate how size and nucleation influence the outcomes of cardiomyocyte cell division in vitro. The Holomonitor M4 is fits within standard cell culture incubators and requires no additional environmental controllers for long-term imaging of living cells.
The interference pattern created between a sample and reference beam generates phase shift data, which is then reconstructed to visualize the cultured cells in three dimensions.

This digital holographic imaging system has several advantages compared to existing live-cell imaging technologies. First, imaging is performed with a low-power 635 nm laser, which minimizes phototoxicity to support long-term imaging of cellular behaviors. Second, full three-dimensional reconstructions are automatically generated for cells located within a 40 μm distance from the cell culture surface, which greatly ameliorates issues with maintaining sample focus during extended imaging durations. And third, the Holomonitor App Suite analysis software enables quantitative tracking of cell size, shape, and movement with single-cell resolution. Using this approach, we explore the interrelationship between cardiomyocyte size, nucleation, and cell cycle control to better understand the cellular mechanisms that drive the loss of mammalian cardiac regenerative capacity after birth.

Min kommentar
Återigen ett bevis för instrumentets förträfflighet att använda inom hjärtforskningen. Forskarna från San Jose öppnade dörren för HoloMonitor inom hjärtstuder i Maj månad och nu börjar vi se resultat.Denna studie lär synas i sömmarna av världens hjärtforskare med ett förmodat medföljande intresse för HoloMonitor.

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Israeliska forskare från HoloMonitoranvändande universitetet The Hebrew University of Jerusalem  har studerat hur munbakterier kan påskynda utveckling av en elak bukspottkörtelcancer.
Studien är tyvärr hårt inlåst,men studiens Supplementary Data visar 7 filmer tagna med HoloMonitor. PHI har själva lagt upp studien på sin hemsida.

Oral bacteria accelerate pancreatic cancer development in mice

Abstract

Objective Epidemiological studies highlight an association between pancreatic ductal adenocarcinoma (PDAC) and oral carriage of the anaerobic bacterium Porphyromonas gingivalis, a species highly linked to periodontal disease. We analysed the potential for P. gingivalis to promote pancreatic cancer development in an animal model and probed underlying mechanisms.

Min kommentar
En ovanlig studie må jag säga. Conclusion: Glöm inte att borsta tänderna morgon o kväll. 😎

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Thailändska forskare har genomlyst användande av inkubatorer vid Long Term Live Cell studies av 2 olika cancerceller.

CO2-Free On-Stage Incubator for Live Cell Imaging of Cholangiocarcinoma Cell Migration on Microfluidic Device

Abstract

Long-term live cell imaging requires sophisticated and fully automated commercial-stage incubators equipped with specified inverted microscopes to regulate temperature, CO2 content, and humidity. In this study, we present a CO2-free on-stage incubator specifically designed for use across various cell culture platforms, enabling live cell imaging applications. A simple and transparent incubator was fabricated from acrylic sheets to be easily placed on the stages of most inverted microscopes. We successfully performed live-cell imaging of cholangiocarcinoma (CCA) cells and HeLa cell dynamics in both 2D and 3D microenvironments over three days. We also analyzed directed cell migration under high serum induction within a microfluidic device. Interesting phenomena such as “whole-colony migration”, “novel type of collective cell migration” and “colony formation during cell and colony migration” are reported here for the first time, to the best of our knowledge. These phenomena may improve our understanding of the nature of cell migration and cancer metastasis.

Studien nämner kortfattat användande av HoloMonitor:  In another study, a ring-shaped culture platform was used for long-term live cell imaging to investigate cell migration and migration speed using an M4 Holomonitor. 

Min kommentar
Man har använt sig av microfluidic device vilket återuppväcker undertecknads intresse för denna teknik.

Får se om jag kan hitta mer aktuell forskning med denna lilla manick.

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Kinesiska forskare har genomfört omfattande studier på den förbannat elaka hjärncancern Glioblastoma.

Dual role of exosomal circCMTM3 derived from GSCs in impeding degradation and promoting phosphorylation of STAT5A to facilitate vasculogenic mimicry formation in glioblastoma

Background

Glioblastoma (GBM), a rare and high-grade malignancy of central nervous system with an extremely high fatality rate in both adults and pediatrics, poses enormous challenges in terms of surgical treatment and adjuvant radiochemotherapy. Despite advancements in biotechnology, our understanding of the pathogenic mechanisms and treatment resistance of GBM has been reassessed, it is still an urgent need to improve clinical outcomes for patients.

Plasticity of cellular phenotypes has been identified as a major contributor to intra- and inter-tumoral heterogeneity as well as treatment resistance in GBM. Increasing evidence indicates that glioblastoma cells exhibit remarkable intrinsic plasticity to acclimate to dynamic microenvironment. Similar to neural stem cells differentiating into neurons and glial cells, GBM stem cells (GSCs) could generate a variety of differentiated morphologies with astrocytic or neuronal phenotypes. Several molecules regulating the switch between GSCs and non-GSCs have been reported, involving nitric oxide promoting the activation of the Notch signaling pathway resulting in the emergence of GSCs phenotypes, and bone morphogenetic protein 4 (BMP4) inducing astroglial-like differentiation and quiescence. Limiting the adaptive capacity of GBM, including targeting plasticity regulators and stemness-differentiation phenotype switching mechanisms, is a worthwhile investigation, which could be therapeutically exploited to overcome treatment resistance.

Cell migration assay

Migration of DGCs was assessed by the HoloMonitor M4 culture system (Phase Holographic Imaging PHI AB, Lund, Sweden). Briefly, cells were seeded into a six-well plate at a density of 10⁴ cells/ml. After cultured and incubated with exosomes, cells were photographed by HoloMonitor M4 culture system every 2 h for a total of 12 h. Cell image of each group was recorded and stored at the last point in time, and then the cellular movement trajectory was simulated and analyzed.

Min kommentar
Kineserna fortsätter sin imponerande kamp mot denna extremt elaka cancer. Jag har skrivit ett flertal inlägg om Glioblastoma och använt starka uttryck för besten. Ordens innehåll speglar inte tillnärmelsevis hur fruktad denna jävla cancer är. Man lever inte länge efter man fått diagnosen. Kineserna ska ha Nobelpriset i medicin om de knäcker den jäveln.Kanske HoloMonitor får ett speciellt omnämnande i sammanhanget? 

Det då instrumentet är så centralt för kinesernas forskning om denna förbannade amöba. Pardon my french.

                                           Mvh the99