Tests by Tissue Culture Methods on the Nature of Immunity Transplanted Skin

1948 ◽  
Vol s3-89 (7) ◽  
pp. 239-252
Author(s):  
P. B. MEDAWAR
Keyword(s):  
Tissue Culture ◽  
Cell Division ◽  
Epidermal Cell ◽  
Cell Suspensions ◽  
Acquired Immunity ◽  
Culture Methods ◽  
Division Frequency

The transplantation of skin from one rabbit to another elicits a reaction that conforms in main outline with that of an actively acquired immunity. The experiments described in this paper were designed to test the hypothesis that the regression of such grafts is secured by the action of antibodies demonstrable in vitro. Skin from adult rabbits has therefore been cultivated in the presence of serum and growing mesenchymal tissues derived solely from rabbits heavily and specifically immunized against it. Immune sera and tissues are without effect on the survival, cell-division frequency and migratory activities of explanted skin, and agglutinins for epidermal cell suspensions are not demonstrable in immune sera. With certain stated qualifications, it has therefore been concluded that the occurrence of free antibodies is not a sufficient explanation of the regression of skin homografts in vivo.

scholarly journals 83ANALYSIS OF RAPID COOLING V. SLOW COOLING COMBINED WITH ICE CRYSTAL SEEDING FOR CRYOPRESERVATION OF PRIMATE EMBRYONIC STEM CELLS

2004 ◽  
Vol 16 (2) ◽  
pp. 163
Author(s):  
S. Baran ◽  
C. Ware
Keyword(s):  
Tissue Culture ◽  
Es Cells ◽  
Embryonic Stem ◽  
Ice Crystal ◽  
Culture Methods ◽  
Slow Cooling ◽  
Rapid Cooling ◽  
Alkaline Phosphatase Staining

Primate embryonic stem (ES) cells have the ability to self-renew indefinitely while maintaining the ability to differentiate. This unique property allows scientists to study the factors necessary for stem cell self-renewal and differentiation in vitro that reflect in vivo processes. Work with primate ES cells is handicapped by the poor survival (1–5%) of rhesus and human ES cells following standard tissue culture methods of rapid cryopreservation. The purpose of this study was to compare and contrast two cryopreservation techniques, slow cooling combined with ice crystal seeding commonly used for mammalian embryos v. rapid cooling commonly used for tissue culture, to find a method for efficient primate ES cell cryopreservation. A combination of trials was run to compare dimethyl sulfoxide (DMSO) v. ethylene glycol as a cryoprotectant, a cooling rate of 0.3°C per minute following ice crystal seeding at −7°C v. placement at −80°C with no seeding, and rapid thaw with step-wise cryoprotectant removal v. one-step sucrose cryoprotectant removal. Cell survival was assessed through a combination of cell surface markers, alkaline phosphatase staining and morphology to look for undifferentiated cells and quantitate survival. All cryopreservations were performed with the same cell density. The survival of the cells with slow embryo-style cooling in DMSO with a step-wise cryoprotectant removal was 64.0% v. 12.8% with rapid cooling.

scholarly journals Application of Horticultural and Tissue Culture Methods for Ex Situ Conservation of Endangered Primula farinosa L.

2020 ◽  
Vol 89 (1) ◽  
Author(s):  
Ewa Sitek ◽  
Barbara Nowak ◽  
Michał Fecowicz ◽  
Zbigniew Gajewski ◽  
Piotr Dańda ◽  
...  
Keyword(s):  
Tissue Culture ◽  
Culture Conditions ◽  
Ex Situ ◽  
In Vitro Cultivation ◽  
Culture Methods ◽  
Transient Nature ◽  
Micropropagated Plants ◽  
The Difference

Our study aimed at active conservation of the last location of <em>Primula farinosa</em>, an endangered species in Poland, and assessed reproduction by seeds and plant propagation on sterile media in tissue culture conditions. We identified gibberellic acid (GA<sub data-id="subscript-1">3</sub>) as the key factor stimulating germination of <em>P. farinosa</em> seeds. Growing juvenile plants under controlled temperature of 18/16 °C day/night yielded good quality plant material without mycorrhization. In tissue culture, the most favorable medium for shoot propagation was MS supplemented with the lowest tested concentration of indole-3-butyric acid (IBA; 0.05 mg dm<sup data-id="superscript-1">−3</sup>) and 6-benzyl-aminopurine (BAP; 0.1 mg dm<sup data-id="superscript-2">−3</sup>). The rooting ability of shoots was high and comparable for all auxins used. 2C DNA content of seed-derived and micropropagated plants did not indicate any change in the ploidy level during in vitro cultivation. Plants derived from seeds and tissue cultures were compared in a 2-year study. Of all the characteristics compared, only the number of flowers per inflorescence was lower for micropropagated plants when compared with the seed-origin plants in the first year of observation. The difference was of transient nature and was not observed in the second year of the study. Effective protocols for in vivo and in vitro propagation of <em>P. farinosa</em> were developed, which can be used in practical species protection.

scholarly journals Necrostatin-1 Supplementation to Islet Tissue Culture Enhances the In-Vitro Development and Graft Function of Young Porcine Islets

2021 ◽  
Vol 22 (16) ◽  
pp. 8367
Author(s):  
Hien Lau ◽  
Shiri Li ◽  
Nicole Corrales ◽  
Samuel Rodriguez ◽  
Mohammadreza Mohammadi ◽  
...  
Keyword(s):  
Tissue Culture ◽  
Oxygen Consumption ◽  
Insulin Secretion ◽  
Oxygen Consumption Rate ◽  
Consumption Rate ◽  
In Vitro Development ◽  
Porcine Islets ◽  
Vitro Development

Pre-weaned porcine islets (PPIs) represent an unlimited source for islet transplantation but are functionally immature. We previously showed that necrostatin-1 (Nec-1) immediately after islet isolation enhanced the in vitro development of PPIs. Here, we examined the impact of Nec-1 on the in vivo function of PPIs after transplantation in diabetic mice. PPIs were isolated from pancreata of 8–15-day-old, pre-weaned pigs and cultured in media alone, or supplemented with Nec-1 (100 µM) on day 0 or on day 3 of culture (n = 5 for each group). On day 7, islet recovery, viability, oxygen consumption rate, insulin content, cellular composition, insulin secretion capacity, and transplant outcomes were evaluated. While islet viability and oxygen consumption rate remained high throughout 7-day tissue culture, Nec-1 supplementation on day 3 significantly improved islet recovery, insulin content, endocrine composition, GLUT2 expression, differentiation potential, proliferation capacity of endocrine cells, and insulin secretion. Adding Nec-1 on day 3 of tissue culture enhanced the islet recovery, proportion of delta cells, beta-cell differentiation and proliferation, and stimulation index. In vivo, this leads to shorter times to normoglycemia, better glycemic control, and higher circulating insulin. Our findings identify the novel time-dependent effects of Nec-1 supplementation on porcine islet quantity and quality prior to transplantation.

scholarly journals SpyAD, a Moonlighting Protein of Group A Streptococcus Contributing to Bacterial Division and Host Cell Adhesion

2014 ◽  
Vol 82 (7) ◽  
pp. 2890-2901 ◽  
Author(s):  
Marilena Gallotta ◽  
Giovanni Gancitano ◽  
Giampiero Pietrocola ◽  
Marirosa Mora ◽  
Alfredo Pezzicoli ◽  
...  
Keyword(s):  
Cell Division ◽  
Mammalian Cells ◽  
Group A Streptococcus ◽  
Host Cells ◽  
Knockout Mutant ◽  
Content Type ◽  
Moonlighting Protein ◽  
Group A

ABSTRACTGroup A streptococcus (GAS) is a human pathogen causing a wide repertoire of mild and severe diseases for which no vaccine is yet available. We recently reported the identification of three protein antigens that in combination conferred wide protection against GAS infection in mice. Here we focused our attention on the characterization of one of these three antigens, Spy0269, a highly conserved, surface-exposed, and immunogenic protein of unknown function. Deletion of thespy0269gene in a GAS M1 isolate resulted in very long bacterial chains, which is indicative of an impaired capacity of the knockout mutant to properly divide. Confocal microscopy and immunoprecipitation experiments demonstrated that the protein was mainly localized at the cell septum and could interactin vitrowith the cell division protein FtsZ, leading us to hypothesize that Spy0269 is a member of the GAS divisome machinery. Predicted structural domains and sequence homologies with known streptococcal adhesins suggested that this antigen could also play a role in mediating GAS interaction with host cells. This hypothesis was confirmed by showing that recombinant Spy0269 could bind to mammalian epithelial cellsin vitroand thatLactococcus lactisexpressing Spy0269 on its cell surface could adhere to mammalian cellsin vitroand to mice nasal mucosain vivo. On the basis of these data, we believe that Spy0269 is involved both in bacterial cell division and in adhesion to host cells and we propose to rename this multifunctional moonlighting protein as SpyAD (StreptococcuspyogenesAdhesion andDivision protein).

scholarly journals Human Adipose-Derived Stromal/Stem Cell Culture and Analysis Methods for Adipose Tissue Modeling In Vitro: A Systematic Review

Cells ◽  
2021 ◽  
Vol 10 (6) ◽  
pp. 1378
Author(s):  
Peyton Gibler ◽  
Jeffrey Gimble ◽  
Katie Hamel ◽  
Emma Rogers ◽  
Michael Henderson ◽  
...  
Keyword(s):  
Systematic Review ◽  
Adipose Tissue ◽  
Drug Discovery ◽  
3D Culture ◽  
Human Adipose Tissue ◽  
Culture Methods ◽  
Adipose Tissue Engineering ◽  
The Impact

Human adipose-derived stromal/stem cells (hASC) are widely used for in vitro modeling of physiologically relevant human adipose tissue. These models are useful for the development of tissue constructs for soft tissue regeneration and 3-dimensional (3D) microphysiological systems (MPS) for drug discovery. In this systematic review, we report on the current state of hASC culture and assessment methods for adipose tissue engineering using 3D MPS. Our search efforts resulted in the identification of 184 independent records, of which 27 were determined to be most relevant to the goals of the present review. Our results demonstrate a lack of consensus on methods for hASC culture and assessment for the production of physiologically relevant in vitro models of human adipose tissue. Few studies have assessed the impact of different 3D culture conditions on hASC adipogenesis. Additionally, there has been a limited use of assays for characterizing the functionality of adipose tissue in vitro. Results from this study suggest the need for more standardized culture methods and further analysis on in vitro tissue functionality. These will be necessary to validate the utility of 3D MPS as an in vitro model to reduce, refine, and replace in vivo experiments in the drug discovery regulatory process.

scholarly journals ELECTRON MICROSCOPY OF PLASMA-CELL TUMORS OF THE MOUSE

1961 ◽  
Vol 9 (2) ◽  
pp. 369-381 ◽  
Author(s):  
D. F. Parsons ◽  
M. A. Bender ◽  
E. B. Darden ◽  
Guthrie T. Pratt ◽  
D. L. Lindsley
Keyword(s):  
Electron Microscopy ◽  
Tissue Culture ◽  
Complex Structure ◽  
Granular Body ◽  
Virus Particles ◽  
Culture Cells ◽  
Large Numbers ◽  
Tumor Agent

The X5563 tumor has been grown in tissue culture. Cells similar to those of the original tumor migrated from the explant and attached to the glass walls of the culture vessels. Electron microscopy showed that large numbers of particles, similar in morphology to virus particles, were associated with these cells after 7 days of culture. The two principal types of particles found in the tumor in vivo appear to be present in vitro. Many more of these particles, however, were larger and showed a more complex structure. Whereas the particles were mainly localized inside endoplasmic reticulum or the Golgi zone in the tumors in vivo, in the tissue culture the majority of the particles were associated with the plasma membrane and were found outside of the cells. The relation of the particles to the granular body is discussed as well as a possible relation to the mammary tumor agent.

In Vivo Interactions with (Tissue Culture Pretreated) Dermal Sheep Collagen

1991 ◽  
Vol 252 ◽  
Author(s):  
P. B. van Wachem ◽  
P. B. van Wachem ◽  
L. H. H. Olde Damink ◽  
P. J. Dijkstra ◽  
J. Feijen ◽  
...  
Keyword(s):  
Tissue Culture ◽  
Cell Death ◽  
Marked Reduction ◽  
Giant Cells ◽  
In Vitro Cytotoxicity ◽  
Cell Infiltration ◽  
Cytotoxic Effects ◽  
Lipid Formation

ABSTRACTPretreatment in tissue culture (TC) was previously found to markedly reduce the in vitro cytotoxicity of two types of crosslinked dermal sheep collagens (DSC's). This in vivo study confirms our in vitro results, in that TC-pretreatment of crosslinked DSC's resulted in the marked reduction or elimination of cytotoxic effects, such as increased cell infiltration, a deviant neutrophil-morphology, lipid formation and cell death. TC-pretreatment affected the crosslinked state of both DSC's in a different way, which could be deduced from the differences in gelatin-formation and presence of giant cells from macrophage- or fibroblast-origin. The results are explained in view of the differences in crosslinking.

scholarly journals Synthesis of Novel Pyran Fragments to Incorporate into Peloruside Analogues

2021 ◽  
Author(s):  
◽  
Oliver Bayley
Keyword(s):  
Cell Division ◽  
Scale Up ◽  
Great Promise ◽  
Peloruside A ◽  
Bioactive Secondary Metabolite ◽  
Mycale Hentscheli ◽  
Significant Attention ◽  
Simple Carbohydrate

<p>Cancer is currently the second largest cause of death globally, leading to a high demand for new and effective chemotherapeutics. For years, natural products have been used as a source of new bioactive compounds; of particular interest in this context, as a source of new chemotherapeutics. One chemotherapeutic candidate which has attracted significant attention in synthetic and medicinal chemistry communities, is peloruside A. Peloruside A is a bioactive secondary metabolite isolated from the New Zealand marine sponge Mycale hentscheli. Since its discovery, peloruside A has shown great promise in cancer studies both in vivo and in vitro with effects observed even at nanomolar concentrations. These chemotherapeutic effects have been shown to occur by halting cell division at the G2/M checkpoint via microtubule stabilisation. Of particular interest is that this stabilisation occurs in a manner distinct from that of the already established taxane class of microtubule stabilising drugs. This means that peloruside A is able to offer both inhibition of cell division in Taxol® resistant cells and synergistic inhibition alongside the current taxane drugs. Since peloruside A is not abundantly available from its natural source, there is a strong incentive for the development of new synthetic strategies for peloruside A production. Unfortunately attempts at aquaculture and attempts at developing an industrial scale synthesis have both proven unsuccessful thus far. In an attempt to overcome some of the difficulties with the scale up of peloruside, analogues have been developed that are intended to have similar bioactivity to peloruside A but simpler, more concise, synthetic routes. These analogues will also enable further elucidation of the binding properties of peloruside A. This project focuses on the generation of a functionalised pyran fragment, starting from a simple carbohydrate, that may be incorporated into the proposed analogues.</p>

scholarly journals Effect of China berry crude extracts on broad mites in vivo and in vitro

2008 ◽  
Vol 2 (1) ◽  
pp. 12-18
Author(s):  
Assma Gatta ◽  
Luaay K. Al – ani ◽  
Nabeel Al - ani
Keyword(s):  
Biological Control ◽  
Tissue Culture ◽  
Melia Azedarach ◽  
Concentration Increase ◽  
Water Extracts ◽  
Crude Extracts ◽  
Number Increase ◽  
Ethanol Extracts

Tissue culture were established from leaf and stem of china berry (Melia azedarach ) tree . Using MS media the best regulator to form callus were 6mg/l BAP, all other concentrations did not give callus . The crude extracts from leaves and callus established from leaves were extracted with water and ethanol with different concentrations. In ethanol extracts the least concentration 0.0001 half of the treated parasites were killed in 24 hours while the number increase as the concentration increase . However in callus the ethanol extracts were much higher about 8.5 were killed in the above concentration . In water extracts the least concentration 0.0001 killed half of the treated parasites in 24 hours .This number was increased 8 or 9 in 48 and 72 hours respectively . These results give us preliminary idea about the biological control of this dangerous parasite.

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