scholarly journals Cell Properties of Lung Tissue-Resident Macrophages Propagated by Co-Culture with Lung Fibroblastic Cells from C57BL/6 and BALB/c Mice

Biomedicines ◽  
2021 ◽  
Vol 9 (9) ◽  
pp. 1241
Author(s):  
Mayu Tsurutani ◽  
Haruka Horie ◽  
Kazushige Ogawa
Keyword(s):  
Lung Tissue ◽  
Culture Media ◽  
Interstitial Cells ◽  
Self Renewal ◽  
Arginase 1 ◽  
Perfused Lung ◽  
Organ Specific ◽  
Standard Culture ◽  
Indispensable Tool ◽  
Fibroblastic Cells

Tissue-resident macrophages (Mø) originating from foetal precursors are maintained by self-renewal under tissue/organ-specific microenvironments (niches). We recently developed a simple propagation method applicable to tissue-resident Mø by co-culturing. Here, we examined the properties of lung tissue-resident Mø propagated by co-culturing with lung interstitial cells. The intracardially and intratracheally perfused lung from BALB/c and C57BL/6 mice could minimise the contamination of alveolar Mø and lung monocytes. Lung tissue-resident Mø could be largely propagated under standard culture media along with the propagation of lung interstitial cells demonstrating a fibroblastic morphology. Propagated lung Mø showed characteristic expression properties for Mø/monocyte markers: high expressions of CD11b, CD64 and CD206; substantial expressions of Mertk; and negative expressions of Ly6C, MHC II and Siglec-F. These properties fit with those of lung interstitial Mø of a certain population that can undergo self-renewal. Propagated fibroblastic cells by co-culturing with lung Mø possessed niche properties such as Csf1 and Tgfb1 expression. Propagated lung Mø from both the mouse types were polarised to an M2 phenotype highly expressing arginase 1 without M2 inducer treatment, whereas the M1 inducers significantly increased the iNOS-positive cell percentages in C57BL/6 mice relative to those in BALB/c mice. This is the first study to demonstrate fundamental properties of lung tissue-resident Mø propagated by co-culturing. Propagated lung Mø showing features of lung interstitial Mø can serve as an indispensable tool for investigating SARS-CoV-2 diseases, although lung interstitial Mø have gained little attention in terms of their involvement in SARS-CoV-2 disease pathology, in contrast to alveolar and recruited Mø.

scholarly journals Simple propagation method for resident macrophages by co-culture and subculture, and their isolation from various organs

2019 ◽  
Vol 20 (1) ◽  
Author(s):  
Kazushige Ogawa ◽  
Mayu Tsurutani ◽  
Aya Hashimoto ◽  
Miharu Soeda
Keyword(s):  
Stromal Cells ◽  
Culture Media ◽  
Expression Patterns ◽  
Adhesive Property ◽  
Optimal Conditions ◽  
Culture Methods ◽  
Simple Method ◽  
Self Renewal ◽  
Standard Culture

Abstract Background Resident macrophages (Mø) originating from yolk sac Mø and/or foetal monocytes colonise tissues/organs during embryonic development. They persist into adulthood by self-renewal at a steady state, independent of adult monocyte inputs, except for those in the intestines and dermis. Thus, many resident Mø can be propagated in vitro under optimal conditions; however, there are no specific in vitro culture methods available for the propagation of resident Mø from diverse tissues/organs. Results We provided a simple method for propagating resident Mø derived from the liver, spleen, lung, and brain of ICR male mice by co-culture and subculture along with the propagation of other stromal cells of the respective organs in standard culture media and successfully demonstrated the propagation of resident Mø colonising these organs. We also proposed a simple method for segregating Mø from stromal cells according to their adhesive property on bacteriological Petri dishes, which enabled the collection of more than 97.6% of the resident Mø from each organ. Expression analyses of conventional Mø markers by flow cytometry showed similar expression patterns among the Mø collected from the organs. Conclusion This is the first study to clearly provide a practical Mø propagation method applicable to resident Mø of diverse tissues and organs. Thus, this novel practical Mø propagation method can offer broad applications for the use of resident Mø of diverse tissues and organs.

Croissance et fructification de Ceratocystis spp. avec attention particulière à C. piceae et C. fimbriata, en fonction du substrat

1982 ◽  
Vol 60 (4) ◽  
pp. 358-363
Author(s):  
A. Thuillier ◽  
P. Neumann
Keyword(s):  
Nutrient Solution ◽  
Culture Media ◽  
Basal Medium ◽  
Malt Agar ◽  
Fruiting Bodies ◽  
Ceratocystis Fimbriata ◽  
Good Production ◽  
Wood Extracts ◽  
Standard Culture ◽  
Better Than

Ceratocystis coerulescens, C. fimbriata, C. ips, and C. minor were tested for production of sexual fruiting bodies, and C. penicillata and C. piceae for asexual fruiting bodies. Ceratocystis fimbriata produced perithecia easily on standard culture media, but there were marked differences between the two strains tested (503, 560). Strain 503 had a good production of fruiting bodies on malt agar (M) and a basal nutrient solution (N). Strain 560 fared better than 503 on Leonian agar (L), but did not fructify on M and N. Supplementing media with various wood extracts produced better results. M + maple sapwood extracts and L + poplar sapwood extracts gave the best results with strain 503, and L + pine sapwood extracts was the best with strain 560.Production of coremia was also influenced by the basal medium and the kind of extracts added as supplements. Fir and maple extracts stimulated the production of fruiting bodies, whereas pine and poplar extracts had no or very little stimulating effects. In every other species tested, the production of fruiting bodies was none or very irregular. [Journal translation]

scholarly journals The effect of Malaysian stingless bee, Trigona spp. honey in promoting proliferation of the undifferentiated stem cell

2019 ◽  
pp. 10-19 ◽  
Author(s):  
Mohd Amin Marwan Mohamad ◽  
Muhammad Alif Mazlan ◽  
Muhammad Ibrahim ◽  
Afzan Mat Yusof ◽  
Shamsul Azlin Ahmad Shamsuddin ◽  
...  
Keyword(s):  
Stem Cells ◽  
Culture Media ◽  
Proliferative Effect ◽  
Natural Medicine ◽  
Ability To Regenerate ◽  
Potential Applications ◽  
Standard Culture ◽  
The Stability ◽  
Dose Dependent ◽  
Standard Culture Medium

Stem cells provide various potential applications in regenerative medicine through its ability of self-renewal and differentiation. Among the various stem cells, dental pulp stem cells (DPSCs) have shown encouraging results in their ability to regenerate. Honey has been used in traditional culture as a natural medicine in supporting wound healing. Yet, very few studies on honey were conducted for its potential as a proliferative agent for stem cells. The aim of this study is to evaluate the stability of two Trigona spp. honeys (1 and 2) added in culture media and its proliferative effect on DPSCs. Both honeys were diluted with standard culture medium through dilution process to prepare the concentrations of 0.01%, 0.04%, 0.10% and 0.25%. DPSCs were treated with the diluted honeys for 24 hours. The proliferative activity was determined through the images taken using an inverted microscope for every six hours. In addition, the MTT assay was conducted to determine the cell viability of DPSCs when treated with both honey 1 and 2 at various concentrations. The results showed a stable culture media added with honey for three days and a dose-dependent proliferative effect of both Trigona spp. honey samples on DPSCs. Optimum proliferative effects were observed at 24 hours for both Trigona spp. honey 1 and 2 on DPSCs. The optimum concentration of Trigona spp. honey 1 was from 0.04% to 0.10% and Trigona spp. honey 2 was below 0.01%. It is concluded that Trigona spp. honey has a promising proliferative effect on DPSCs.

scholarly journals Application of Flow Cytometry to Segregated Kinetic Modeling Based on the Physiological States of Microorganisms

2007 ◽  
Vol 73 (12) ◽  
pp. 3993-4000 ◽  
Author(s):  
Covadonga Quir�s ◽  
M�nica Herrero ◽  
Luis A. Garc�a ◽  
Mario D�az
Keyword(s):  
Flow Cytometry ◽  
Culture Media ◽  
Glucose Consumption ◽  
Viable Cell ◽  
Counting Method ◽  
Batch Cultures ◽  
Viable Cells ◽  
The Status ◽  
Standard Culture

ABSTRACT Flow cytometry (FC) has been introduced to characterize and to assess the physiological states of microorganisms in conjunction with the classical plate-counting method. To show the applicability of the technique, in particular for the development of kinetic models, pure culture fermentation experiments were followed over time, using both prokaryotic (Lactobacillus hilgardii) and eukaryotic (Saccharomyces cerevisiae) microorganisms growing in standard culture media (MRS and YPD). The differences observed between the active and viable cells determined by FC and CFU, respectively, allowed us to determine that a large number of cells were in a viable but nonculturable (VBNC) state, which resulted in a subpopulation much larger than the damaged-cell (double-stained) subpopulation. Finally, the determination of the evolution of viable, the VBNC, and the dead cells allowed us to develop a segregated kinetic model to describe the yeast and the bacteria population dynamics and glucose consumption in batch cultures. This model, more complete than that which is traditionally used, based only on viable cell measurements, describes better the behavior and the functionality of the cultures, giving a deeper knowledge in real time about the status and the course of the bioprocesses.

scholarly journals Open microfluidic coculture reveals paracrine signaling from human kidney epithelial cells promotes kidney specificity of endothelial cells

2020 ◽  
Vol 319 (1) ◽  
pp. F41-F51
Author(s):  
Tianzi Zhang ◽  
Daniel Lih ◽  
Ryan J. Nagao ◽  
Jun Xue ◽  
Erwin Berthier ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Epithelial Cells ◽  
Matrix Metalloproteinase ◽  
Culture Media ◽  
Human Kidney ◽  
Organ Specificity ◽  
Paracrine Signaling ◽  
Human Organ ◽  
Extrinsic Cues ◽  
Organ Specific

Endothelial cells (ECs) from different human organs possess organ-specific characteristics that support specific tissue regeneration and organ development. EC specificity is identified by both intrinsic and extrinsic cues, among which the parenchyma and organ-specific microenvironment are critical contributors. These extrinsic cues are, however, largely lost during ex vivo cultures. Outstanding challenges remain to understand and reestablish EC organ specificity for in vitro studies to recapitulate human organ-specific physiology. Here, we designed an open microfluidic platform to study the role of human kidney tubular epithelial cells in supporting EC specificity. The platform consists of two independent cell culture regions segregated with a half wall; culture media are added to connect the two culture regions at a desired time point, and signaling molecules can travel across the half wall (paracrine signaling). Specifically, we report that in the microscale coculture device, primary human kidney proximal tubule epithelial cells (HPTECs) rescued primary human kidney peritubular microvascular EC (HKMEC) monolayer integrity and fenestra formation and that HPTECs upregulated key HKMEC kidney-specific genes (hepatocyte nuclear factor 1 homeobox B, adherens junctions-associated protein 1, and potassium voltage-gated channel subfamily J member 16) and endothelial activation genes (vascular cell adhesion molecule-1, matrix metalloproteinase-7, and matrix metalloproteinase-10) in coculture. Coculturing with HPTECs also promoted kidney-specific genotype expression in human umbilical vein ECs and human pluripotent stem cell-derived ECs. Compared with culture in HPTEC conditioned media, coculture of ECs with HPTECs showed increased upregulation of kidney-specific genes, suggesting potential bidirectional paracrine signaling. Importantly, our device is compatible with standard pipettes, incubators, and imaging readouts and could also be easily adapted to study cell signaling between other rare or sensitive cells.

scholarly journals Microscopy and Microanalysis of an Extreme Case of Salt and Biodegradation in 17th Century Wall Paintings

2015 ◽  
Vol 21 (3) ◽  
pp. 606-616 ◽  
Author(s):  
Milene Gil ◽  
Maria Rosário Martins ◽  
Maria Luisa Carvalho ◽  
Cátia Souto ◽  
Stephane Longelin ◽  
...  
Keyword(s):  
Culture Media ◽  
Extreme Case ◽  
Biological Assessment ◽  
Salt Weathering ◽  
Bacterial Strains ◽  
Isolation And Identification ◽  
Mineralogical Characterization ◽  
Fungal Isolation ◽  
X Ray ◽  
Standard Culture

AbstractThe present study characterizes the main deterioration mechanisms affecting the early 17th frescoes of Casa de Fresco, the only known example in Portugal of a semi-underground leisure room richly decorated with a balcony over a water well. Frescoes from the vault are at risk due to salt weathering and biodeterioration. The aim of the research was identification of the deterioration materials, determination of their origin, and their effect on the frescoes before future intervention. Scanning electron microscopy with an energy-dispersive X-ray detector (SEM-EDS) was used to determine salt morphology and microanalysis. The mineralogical characterization was performed by X-ray powder diffraction, complemented with µ-Raman and µ-Fourier transform infrared spectroscopy. Biological assessment was evaluated with optical microscopy and SEM-EDS. Bacterial and fungal isolation and identification were performed using standard culture media and methods according to Bergey’s Manual of Systematic Bacteriology and from the Compendium of Soil Fungi. The results show that Ca and Ca-Mg carbonates from the paint renderings are the predominant salt species affecting the site. Bacterial strains from the genera Bacillus and Pseudomonas and fungal strains from the Cladosporium spp. and Penicillium spp. were isolated in the salt formations, within and between the mortar layers. Azurite, malachite, and smalt paint layers are the most affected by the weathering conditions.

scholarly journals PROMOTION OF REPLICATION IN LYMPHOID CELLS BY SPECIFIC THIOLS AND DISULFIDES IN VITRO

1973 ◽  
Vol 138 (3) ◽  
pp. 574-592 ◽  
Author(s):  
J. D. Broome ◽  
M. W. Jeng
Keyword(s):  
Culture Media ◽  
Growth Promotion ◽  
Lymphoid Cells ◽  
Splenic Lymphocytes ◽  
Mitogenic Response ◽  
Lymphoma Cells ◽  
Structure Activity ◽  
Standard Culture ◽  
Increased Sensitivity

Numerous lines of mouse lymphoid tumors (13 of 22 tested) showed, with increased sensitivity, a property of normal mouse splenic lymphocytes, the potential for growth promotion in vitro by specific thiols added to standard culture media. For lymphoma L1210 (V), structure activity relationships were examined; 9 of 30 thiols promoted growth; the most active was α-thioglycerol, effective at 0.2 µM. Thiols became oxidized under conditions of tissue culture and had half-lives of less than 8 h. Disulfides of active thiols promoted growth of lymphoma cells. The mitogenic response of splenic lymphocytes to lectins was increased by thiols-disulfides which promoted the growth of lymphoma cells, but the response varied with the mitogen preparation used and under some conditions thiols-disulfides were inhibitory.

scholarly journals Morphological and Molecular Characterization of Phoma complanata, a New Causal Agent of Archangelica officinalis Hoffm. in Poland

2017 ◽  
Vol 66 (2) ◽  
pp. 281-285 ◽  
Author(s):  
Beata Zimowska ◽  
Ewa Dorota Zalewska ◽  
Ewa Dorota Król ◽  
Agnieszka Furmańczyk
Keyword(s):  
Culture Media ◽  
Culture Collection ◽  
Morphological Similarity ◽  
Reference Isolate ◽  
Angelica Archangelica ◽  
Its Regions ◽  
Standard Culture ◽  
First Time ◽  
Morphological And Molecular Characterization

The paper concerns the fungus Phoma complanata, isolated for the first time in Poland, from the roots and umbels of angelica (Archangelica officinalis) in 2009. The morphology of fungal isolates was tested on standard culture media. Moreover, the sequence analysis of ITS regions was conducted. Morphological similarity of P. complanata Polish isolates to the reference isolate obtained from CBS culture collection was determined and together with the molecular analysis confirmed the affiliation of the fungus to the species.

scholarly journals Autoimmune reactions in the bronchial asthma in children induced heterophilic antigenes of microorganism of bronchopulmants

2019 ◽  
Keyword(s):  
Bronchial Asthma ◽  
Lung Tissue ◽  
Cellular Tissue ◽  
Pathological Process ◽  
Quantitative Calculation ◽  
Stage Of Development ◽  
Cell Tissue ◽  
Organ Specific ◽  
Water Salt ◽  
Tissue Structures

Introduction. At the present stage of development of the problem in the etiopathogenesis of bronchial asthma (BA) in children, regardless of form, one of the leading places belongs to the microbial factor. Aim. The aim of the work was to study the development of autoimmune reactions to the cellular tissue structures of the trachea, bronchi and lung tissue, stimulated by heterophilic antigens of the microbiota of the bronchopulmonary system of children with BA. Materials and methods. A total of 97 children with BA aged 7 to 15 years were examined. The diagnosis of the disease was established according to GINA (2017) and the order of the Ministry of Health of Ukraine dated 08.10.2013 No. 868. Heterophilic antigens of bronchopulmonary structures in microbiota were determined using hyperimmune organ-specific rabbit sera to antigens of the trachea, bronchi and lung tissue. Lipopolysaccharide antigens from homologous cell-tissue structures of the trachea, pulmonary bronchi were determined, water-salt antigens from the structures of the trachea, bronchi, and lung tissue were obtained from accidentally dead children with I (0) blood group. The level of autoantibodies to antigens of the bronchopulmonary system with the quantitative calculation of the indicator Qφ was determined in the nephelometric reaction. Results. In the work it was shown experimentally that microorganisms, isolated from sputum of children, patients with asthma in the period of exacerbation, varying their antigenic potential, are able to include in their structure heterophilic antigens of cell-tissue structures of the bronchopulmonary system. Microorganisms including in their structure heterophilic antigens of the trachea, bronchi and lung tissue not only determine the induction of the pathological process in the bronchopulmonary system, but also translate it into an autoimmune basis, exacerbating the severity of the course of the disease. Conclusions. The study showed that the proposed methods are important for clarifying the etiopathogenesis of BA in children and disclosing the mechanism for switching the pathological process in the bronchopulmonary system to an autoimmune basis and can be used to develop new approaches for the etiopathogenetic treatment of the disease.

scholarly journals SALL4 and microRNA: The Role of Let-7

Genes ◽  
2021 ◽  
Vol 12 (9) ◽  
pp. 1301
Author(s):  
Jun Liu ◽  
Madeline A. Sauer ◽  
Shaza Hussein ◽  
Junyu Yang ◽  
Daniel G. Tenen ◽  
...  
Keyword(s):  
Stem Cells ◽  
Embryonic Stem ◽  
Mammalian Development ◽  
Multiple Cancer ◽  
Self Renewal ◽  
Zinc Finger Transcription Factor ◽  
Different Types ◽  
Organ Specific ◽  
Cancer Types

SALL4 is a zinc finger transcription factor that belongs to the spalt-like (SALL) gene fam-ily. It plays important roles in the maintenance of self-renewal and pluripotency of embryonic stem cells, and its expression is repressed in most adult organs. SALL4 re-expression has been observed in different types of human cancers, and dysregulation of SALL4 contributes to the pathogenesis, metastasis, and even drug resistance of multiple cancer types. Surprisingly, little is known regard-ing how SALL4 expression is controlled, but recently microRNAs (miRNAs) have emerged as im-portant regulators of SALL4. Due to the ability of regulating targets differentially in specific tissues, and recent advances in systemic and organ specific miRNA delivery mechanisms, miRNAs have emerged as promising therapeutic targets for cancer treatment. In this review, we summarize cur-rent knowledge of the interaction between SALL4 and miRNAs in mammalian development and cancer, paying particular attention to the emerging roles of the Let-7/Lin28 axis. In addition, we discuss the therapeutic prospects of targeting SALL4 using miRNA-based strategies, with a focus on the Let-7/LIN28 axis.

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