The Behaviour in vitro of Dissociated Embryonic Pituitary Tissue

Development ◽  
1958 ◽  
Vol 6 (3) ◽  
pp. 518-526
Author(s):  
H. Sobel
Keyword(s):  
Secretory Granules ◽  
Anterior Lobe ◽  
Cellular Level ◽  
Chick Embryos ◽  
Histological Differentiation ◽  
Limb Buds ◽  
Discrete State ◽  
Viable Cells ◽  
Cultivation In Vitro

It was previously reported (A. Moscona & H. Moscona, 1952) that the tissues of limb-buds and mesonephroi of early chick embryos can be dissociated into suspensions of discrete viable cells which, under certain conditions of cultivation in vitro, reaggregate into clusters and re-establish a tissue-like association. Upon further cultivation in vitro these primary cellular associations became transformed into organized tissue patterns, the development of which proceeds to the level of typical histological differentiation. Owing to the nature of the experimental material studied so far, it has mainly been the capacity of the aggregates for re-establishing typical intercellular relationship that has come prominently into view. The present observations were aimed at examining the capacity of cells, aggregated from a discrete state, to resume and complete differentiation on the cellular level, e.g. to achieve a cytologically characteristic secretory status. The normally developed cells of the anterior lobe of the pituitary carry a distinct mark of their state of differentiation—the secretory granules.

scholarly journals A PURE STRAIN OF CARTILAGE CELLS IN VITRO

1922 ◽  
Vol 36 (4) ◽  
pp. 379-384 ◽  
Author(s):  
Albert Fischer
Keyword(s):  
Close Contact ◽  
Initial Growth ◽  
Chick Embryos ◽  
Small Lymphocyte ◽  
Pure Strain ◽  
Thin Membranes ◽  
Cartilage Cells ◽  
Cultivation In Vitro ◽  
Large Nucleolus

1. A strain of cartilage cells, obtained from the pars cartilago scleræ of the eye of chick embryos, has been cultivated for more than 3 months in vitro. 2. The initial growth of the cartilage was possible only on the free surface of the coagulum. 3. The hyaline substance disappeared during cultivation in vitro. The succeeding stages of a transformation from small, lymphocyte-like cells into large, spindle-shaped cells were observed. The cartilage cells were spindle-shaped and grew in close contact, forming thin membranes. In surface-grown cartilage cells, the nucleus, usually containing one large nucleolus, stained less deeply than the cytoplasm. 4. The rate of growth of cartilage was slower than that of fibroblasts and epithelium. After cultivation on the surface of the coagulum, the cartilage cells could multiply even when embedded in the coagulum. But their growth was less extensive and uniform.

scholarly journals CULTURES OF ORGANIZED TISSUES

1922 ◽  
Vol 36 (4) ◽  
pp. 393-397 ◽  
Author(s):  
Albert Fischer
Keyword(s):  
Small Body ◽  
Chick Embryos ◽  
Malignant Cells ◽  
Intestinal Tissue ◽  
Fluid Medium ◽  
Long Period ◽  
Muscle Tissues ◽  
Pure Cultures ◽  
Cultivation In Vitro

An artificial organism, if one may so term it, composed of a complex of tissues, was cultivated for a long period of time. Small fragments of intestine from chick embryos 20 to 21 days old were placed in a suitable medium. The epithelium proliferated and completely covered the fragment of intestine after 4 to 6 days. A small body was thus formed, round or oblong in shape, surrounded by cylindrical epithelium and containing epithelial, connective, and muscle tissues, endothelium, and ameboid cells. After a month's cultivation in vitro, no necrosis had occurred. Therefore, it may be assumed that, through the intestinal epithelium, the medium supplied the intestinal tissue with sufficient nourishment. No uncontrolled proliferation took place after the epithelium bad surrounded the entire fragment. The cultivation of complex tissues will facilitate the study of the interactions of the different cells under various conditions. In some experiments, pure cultures of epithelial cells were grafted into such an "organism" without difficulty. The growth of malignant cells could be studied in the same way. When the "organism" was placed in a fluid medium, the epithelium remained normal but the stroma disappeared. It seems that plasma played an important rôle in the maintenance of the tissues in their normal condition.

scholarly journals EFFECT OF LIVER AND PITUITARY DIGESTS ON THE PROLIFERATION OF SARCOMATOUS FIBROBLASTS OF THE RAT

1928 ◽  
Vol 47 (3) ◽  
pp. 371-378 ◽  
Author(s):  
Lillian E. Baker ◽  
Alexis Carrel
Keyword(s):  
Chick Embryo ◽  
Pituitary Gland ◽  
Proliferative Activity ◽  
Fatty Degeneration ◽  
Anterior Lobe ◽  
Pure Strain ◽  
Cultivation In Vitro

1. A media containing all the essential constituents for the cultivation in vitro of sarcomatous fibroblasts of the rat has been prepared by digesting calf liver and also the anterior lobe of the pituitary body with pepsin. 2. The nutritive action of the pituitary digests is not altered by thorough extraction with ether. 3. After a pure strain of sarcomatous fibroblasts had been cultivated for 3 months in a liver digest, its proliferative activity was as great as at the beginning of the experiment. The same was true of the colonies cultivated for 1 month in a digest of pituitary gland. The increase in the volume of the colonies which takes place in the digests is about as great as that produced by chick embryo juice. 4. Normal chicken fibroblasts also proliferate in both digests, but they undergo fatty degeneration after a more or less prolonged period of cultivation.

Ultrastructural aspects of early development of the fore-limb buds in the chick and the mouse

1965 ◽  
Vol 162 (988) ◽  
pp. 387-405 ◽  
Keyword(s):  
Endoplasmic Reticulum ◽  
Early Development ◽  
Secretory Granules ◽  
Free Cell ◽  
Mouse Embryos ◽  
Limb Bud ◽  
Chick Embryos ◽  
Limb Buds ◽  
Mouse Limb ◽  
A Chain

Light microscope investigations of the early development of the fore-limb buds in chick and mouse were made to guide electron microscope studies with these tissues. At the time of maximal development of the ectodermal apical ridge there is a higher concentration of cytoplasmic RNA in the apical ridge cells than in the other cells of the limb bud. Ultrastructural investigations showed that, in the mesoblast cells at the earliest stages, profiles of endoplasmic reticulum are often found attached to the outer nuclear membrane. Some what later, discontinuities of nuclear envelope occur by which the content of the nucleus may communicate with the endoplasmic reticulum. In the cytoplasm of the mesoblast cells at these stages there were many granules similar in form and size to secretory granules of gland cells. Ribosomes are in the polysomal condition. At stages later than 20 in chick and in 11-day-old mouse embryos, the mesoblast shows the character of a syncytial tissue. Epiblast cells possess all the characters of an epithelium with well developed junctional complexes. The desmosomes form a chain consisting of units equipped with individual dense plaques, but connected by continuous bundles of fibres running parallel to the chain. The free cell membrane of the epiblastic cells, particularly at early stages, forms numerous microvilli and single cilia. In later stages during the form action of the ectodermal apical ridge, cilia have been found between the cells. This fact indicates that when the apical ridge is formed ectodermal cells migrate towards the margin of the limb bud. At these stages microvilli are also found between the apical ridge cells where they contribute to the cell-to-cell adhesion. Beginning at stage 22 in chick embryos and from the 12th day in mouse embryos there are in cells of the apical ridge numerous and extensive Golgi systems spread throughout the cytoplasm. Some what later there appear successively lysosomes, cytolysomes and extranuclear necrotic centres. All these organelles manifest acid phosphatase activity and are thoughtto initiate the involutive process in the apical ridge. Pycnosis and karyorrhexis appear as the last stage of cellular degeneration. Degenerating cells undergo phagocytosis by neighbouring epithelial cells. A basement membrane is present at all stages of development of the chick and mouse limb buds. It is an acellular continuous structure lining the internal (basal) surface of the epiblast, but in chick embryos it shows discontinuities immediately under the apical ectodermal ridge at the time of its maximum development.

IGF-I, insulin and FGFs induce outgrowth of the limb buds of amelic mutant chick embryos

Development ◽  
1996 ◽  
Vol 122 (4) ◽  
pp. 1323-1330 ◽  
Author(s):  
C.N. Dealy ◽  
R.A. Kosher
Keyword(s):  
Proper Time ◽  
Limb Bud ◽  
Chick Embryos ◽  
In Vitro Morphogenesis ◽  
Limb Buds ◽  
Limb Outgrowth ◽  
Igf I ◽  
High Level

IGF-I, insulin, FGF-2 and FGF-4 have been implicated in the reciprocal interactions between the apical ectodermal ridge (AER) and underlying mesoderm required for outgrowth and patterning of the developing limb. To study further the roles of these growth factors in limb outgrowth, we have examined their effects on the in vitro morphogenesis of limb buds of the amelic mutant chick embryos wingless (wl) and limbless (ll). Limb buds of wl and ll mutant embryos form at the proper time in development, but fail to undergo further outgrowth and subsequently degenerate. Wl and ll limb buds lack thickened AERs capable of promoting limb outgrowth, and their thin apical ectoderms fail to express the homeobox-containing gene Msx-2, which is highly expressed by normal AERs and has been implicated in regulating AER activity. Here we report that exogenous IGF-I and insulin, and, to a lesser extent, FGF-2 and FGF-4 induce the proliferation and directed outgrowth of explanted wl and ll mutant limb buds, which in vitro, like in vivo, normally fail to undergo outgrowth and degenerate. IGF-I and insulin, but not FGFs, also cause the thin apical ectoderms of wl and ll limb buds to thicken and form structures that grossly resemble normal AERs and, moreover, induce high level expression of Msx-2 in these thickened AER-like structures. Neither IGF-I, insulin nor FGFs induce expression of the homeobox-containing gene Msx-1 in the subapical mesoderm of wl or ll limb buds, although FGFs, but not IGF-I or insulin, maintain Msx-1 expression in normal (non-mutant) limb bud explants lacking an AER. The implications of these results to the relationships among the wl and ll genes, IGF-I/insulin, FGFs, Msx-2 and Msx-1 in the regulation of limb outgrowth is discussed.

Endogenous Viable Cells in Lyopreserved Amnion Retain Differentiation Potential and Anti-Fibrotic Activity In Vitro

2019 ◽  
Author(s):  
Yong Mao ◽  
Tyler Hoffman ◽  
Sandeep Dhall ◽  
Amit Singal ◽  
Malathi Sathyamoorthy ◽  
...  
Keyword(s):  
Differentiation Potential ◽  
Viable Cells

Exploiting Anti-Inflammation Effects of Flavonoids in Chronic Inflammatory Diseases

2020 ◽  
Vol 26 (22) ◽  
pp. 2610-2619 ◽  
Author(s):  
Tarique Hussain ◽  
Ghulam Murtaza ◽  
Huansheng Yang ◽  
Muhammad S. Kalhoro ◽  
Dildar H. Kalhoro
Keyword(s):  
Oxidative Stress ◽  
Chronic Diseases ◽  
Inflammatory Diseases ◽  
Therapeutic Option ◽  
Cellular Level ◽  
Antiinflammatory Drugs ◽  
Suitable Alternative ◽  
Chronic Inflammatory Diseases

Background: Inflammation is a complex response of the host defense system to different internal and external stimuli. It is believed that persistent inflammation may lead to chronic inflammatory diseases such as, inflammatory bowel disease, neurological and cardiovascular diseases. Oxidative stress is the main factor responsible for the augmentation of inflammation via various molecular pathways. Therefore, alleviating oxidative stress is effective a therapeutic option against chronic inflammatory diseases. Methods: This review article extends the knowledge of the regulatory mechanisms of flavonoids targeting inflammatory pathways in chronic diseases, which would be the best approach for the development of suitable therapeutic agents against chronic diseases. Results: Since the inflammatory response is initiated by numerous signaling molecules like NF-κB, MAPK, and Arachidonic acid pathways, their encountering function can be evaluated with the activation of Nrf2 pathway, a promising approach to inhibit/prevent chronic inflammatory diseases by flavonoids. Over the last few decades, flavonoids drew much attention as a potent alternative therapeutic agent. Recent clinical evidence has shown significant impacts of flavonoids on chronic diseases in different in-vivo and in-vitro models. Conclusion: Flavonoid compounds can interact with chronic inflammatory diseases at the cellular level and modulate the response of protein pathways. A promising approach is needed to overlook suitable alternative compounds providing more therapeutic efficacy and exerting fewer side effects than commercially available antiinflammatory drugs.

TNF-α Suppresses Autophagic Flux in Acinar Cells in IgG4-Related Sialadenitis

2019 ◽  
Vol 98 (12) ◽  
pp. 1386-1396 ◽  
Author(s):  
X. Hong ◽  
S.N. Min ◽  
Y.Y. Zhang ◽  
Y.T. Lin ◽  
F. Wang ◽  
...  
Keyword(s):  
Acinar Cell ◽  
Cell Injury ◽  
Ex Vivo ◽  
Secretory Granules ◽  
Acinar Cells ◽  
Autophagic Flux ◽  
C6 Cells ◽  
Tnf Α ◽  
Α Level

IgG4-related sialadenitis (IgG4-RS) is a newly recognized immune-mediated systemic fibroinflammatory disease that affects salivary glands and leads to hyposalivation. Tumor necrosis factor–α (TNF-α) is a critical proinflammatory cytokine involved in several salivary gland disorders, but its role and mechanism regarding acinar cell injury in IgG4-RS are unknown. Here, we found that TNF-α level was significantly increased in serum and submandibular gland (SMG) of patients and that serum TNF-α level was negatively correlated with saliva flow rate. Ultrastructural observations of IgG4-RS SMGs revealed accumulation of large autophagic vacuoles, as well as dense fibrous bundles, decreased secretory granules, widened intercellular spaces, swollen mitochondria, and expanded endoplasmic reticulum. Expression levels of LC3 and p62 were both increased in patients’ SMGs. TNF-α treatment led to elevated levels of LC3II and p62 in both SMG-C6 cells and cultured human SMG tissues but did not further increase their levels when combined with bafilomycin A1 treatment. Moreover, transfection of Ad-mCherry-GFP-LC3B in SMG-C6 cells confirmed the suppression of autophagic flux after TNF-α treatment. Immunofluorescence imaging revealed that costaining of LC3 and the lysosomal marker LAMP2 was significantly decreased in patients, TNF-α–treated SMG-C6 cells, and cultured human SMGs, indicating a reduction in autophagosome-lysosome fusion. Furthermore, the ratio of pro/mature cathepsin D was elevated in vivo, ex vivo, and in vitro. TNF-α also appeared to induce abnormal acidification of lysosomes in acinar cells, as assessed by lysosomal pH and LysoTracker DND-26 fluorescence intensity. In addition, TNF-α treatment induced transcription factor EB (TFEB) redistribution in SMG-C6 cells, which was consistent with the changes observed in IgG4-RS patients. TNF-α increased the phosphorylation of extracellular signal–regulated kinase (ERK) 1/2, and inhibition of ERK1/2 by U0126 reversed TNF-α–induced TFEB redistribution, lysosomal dysfunction, and autophagic flux suppression. These findings suggest that TNF-α is a key cytokine related to acinar cell injury in IgG4-RS through ERK1/2-mediated autophagic flux suppression.

scholarly journals Investigation of Chlorella pyrenoidosa Protein as a Source of Novel Angiotensin I-Converting Enzyme (ACE) and Dipeptidyl Peptidase-IV (DPP-IV) Inhibitory Peptides

Nutrients ◽  
2021 ◽  
Vol 13 (5) ◽  
pp. 1624
Author(s):  
Yuchen Li ◽  
Gilda Aiello ◽  
Enrico Mario Alessandro Fassi ◽  
Giovanna Boschin ◽  
Martina Bartolomei ◽  
...  
Keyword(s):  
Chlorella Pyrenoidosa ◽  
Cellular Level ◽  
Potential Interaction ◽  
Converting Enzyme ◽  
Inhibitory Peptides ◽  
Angiotensin I Converting Enzyme ◽  
Dpp Iv ◽  
Ace Activity ◽  
The Stability

Chlorella pyrenoidosa (C. pyrenoidosa) is a microalgae species with a remarkably high protein content that may potentially become a source of hypotensive and hypoglycemic peptides. In this study, C. pyrenoidosa proteins were extracted and hydrolyzed overnight with pepsin and trypsin with final degrees of hydrolysis of 18.7% and 35.5%, respectively. By LC-MS/MS, 47 valid peptides were identified in the peptic hydrolysate (CP) and 66 in the tryptic one (CT). At the concentration of 1.0 mg/mL, CP and CT hydrolysates inhibit in vitro the angiotensin-converting enzyme (ACE) activity by 84.2 ± 0.37% and 78.6 ± 1.7%, respectively, whereas, tested at cellular level at the concentration of 5.0 mg/mL, they reduce the ACE activity by 61.5 ± 7.7% and 69.9 ± 0.8%, respectively. At the concentration of 5.0 mg/mL, they decrease in vitro the DPP-IV activity by 63.7% and 69.6% and in Caco-2 cells by 38.4% and 42.5%, respectively. Short peptides (≤10 amino acids) were selected for investigating the potential interaction with ACE and DPP-IV by using molecular modeling approaches and four peptides were predicted to block both enzymes. Finally, the stability of these peptides was investigated against gastrointestinal digestion.

scholarly journals Bactericidal activity of three different antiseptic ophthalmic preparations as surgical prophylaxis

2021 ◽  
Author(s):  
Daniele Tognetto ◽  
Marco R. Pastore ◽  
Gian Marco Guerin ◽  
Giuliana Decorti ◽  
Martina Franzin ◽  
...  
Keyword(s):  
Antimicrobial Activity ◽  
Povidone Iodine ◽  
Bacterial Load ◽  
Viable Cells ◽  
Polyethylene Glycol 1000 ◽  
Oil Concentration ◽  
Short Time ◽  
First Time ◽  
Antiseptic Solutions

Abstract Purpose In the era of antibiotic resistance, there is an increased interest in antiseptic solutions that might represent a reliable option for ocular surface disinfection. The objective of this study is to compare for the first time three different antiseptic ophthalmic preparations to assess their in vitro antimicrobial activity. Methods The antiseptic activity of three commercial ophthalmic solutions, IODIM (povidone-iodine 0.6% in hyaluronic acid vehicle—Medivis, Catania, Italy), OZODROP (nanoemulsion with ozonated oil—concentration not specified—FBVision, Ophthalmic Pharmaceuticals, Rome, Italy), and DROPSEPT (chlorhexidine 0.02% and vitamin E 0.5% Tocopherol Polyethylene Glycol 1000 Succinate—TPGS, Sooft Italia, Montegiorgio, Italy), was tested in vitro on six reference strains by time-killing assays. Viable cells were evaluated after 1, 15, 30 min; 2, 6, and 24 h exposure by seeding 100 µl of the suspension (or appropriate dilutions) on LB agar or Sabouraud-dextrose agar. All plates were incubated at 37 °C for 24 h and evaluated by manually counting the colonies. Results IODIM solution showed a very rapid microbicidal activity: the number of viable cells for all the tested strains was under the detection limit (less than 10 CFU/ml) already after 1 min exposure, and this result was maintained at every incubation time. The rapid antimicrobial activity of povidone-iodine was not replicated when testing the other two antiseptics. Conclusions The study reports the great efficacy in reducing bacterial load in a very short time of povidone-iodine 0.6% compared with other antiseptic preparations.

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