PSX-B-16 Recovery of live cells from refrigerated sheep skin after different days of cold storage

2021 ◽  
Vol 99 (Supplement_3) ◽  
pp. 328-329
Author(s):  
Mahipal Singh ◽  
Xiaoling Ma
Keyword(s):  
Storage Time ◽  
Karyotype Analysis ◽  
Plasmid Vector ◽  
Live Cells ◽  
Time Interval ◽  
Normal Female ◽  
Primary Cells ◽  
Growth Morphology ◽  
Gfp Gene ◽  
Similar Growth

Abstract Cryopreservation of tissues from domesticated and wild relatives has been suggested to conserve genetic diversity. Ensuring that the tissues have live cells prior to preservation, especially in postmortem tissues, is an essential stem for success. How long cells live after clinical death is not precisely known in animals. The objective of this study was to evaluate the limits of cell survival in sheep skin stored at 4°C postmortem. Ear skin was procured from six random but healthy slaughtered animals and stored at 4°C in the lab. Ten explants (2–3 mm2) were cultured from each animal in DMEM media with 10% FBS, 50 units/mL of penicillin, 50 µg/mL of streptomycin, and 2.5 µg/mL of fungizone on two 60 mm dishes after 0, 10, 20, 27, 30, 35, 38, 41, 45, 50, 55, 60, 65 and 70 days of storage. Outgrowth of fibroblast-like cells around the explants was scored after 10 days of culture in a CO2 incubator. Results show outgrowth of cells up to 65 days of postmortem storage. Out of 476 explants adhered to dish surface, 374 (78.58%) exhibited outgrowth. The number of outgrowing cells decreased with increasing postmortem storage time interval. To test the differences between cell cultures obtained from postmortem fresh and stored tissues, we established secondary cultures from primary cells of 0-dpm and 65-dpm time points from selected cell lines. Both cultures exhibited similar growth morphology and growth curve, could be cryopreserved with >80% post freezing cell viability, lasted in cultures up to 35 passages, and expressed GFP gene upon transfection with a GFP gene containing plasmid vector. The karyotype analysis of 65-dpm tissue derived cells revealed a normal female karyotype without any genetic aberrations. These results suggest that normal proliferative cells can be recovered from sheep skin up to about 2 months postmortem, if kept refrigerated.

scholarly journals Postmortem recovery of proliferative fibroblasts up to three days in livestock ear skin stored at 35°C

2017 ◽  
Vol 9 (2) ◽  
pp. 9
Author(s):  
Mahipal Singh ◽  
Venkata N. Degala
Keyword(s):  
Elevated Temperature ◽  
Normal Growth ◽  
Postmortem Interval ◽  
Time Interval ◽  
Normal Female ◽  
Primary Cells ◽  
Fresh Tissue ◽  
Cell Therapies ◽  
Skin Explants ◽  
Female Sheep

Effect of elevated temperature (35°C) on postmortem survival of cells in goat, sheep and bovine tissues was studied. Skin explants (n = 30; 2-3 mm2) were cultured in petri dishes after 2, 24, 48, 72, 96 and 120 hours of postmortem interval in DMEM media supplemented with 10% FBS, 50 units/mL of penicillin, 50 µg/mL of streptomycin and 2.5 µg/mL of fungizone. Outgrowth of cells around the explants was observed up to 72 h in sheep, 48 h in bovine and 24 h in goats. In general, the number of explants exhibiting outgrowth as well as the level of confluence decreased with increasing postmortem time interval. Secondary cultures established from primary cells for 72 h postmortem interval show cytogenetically stable chromosomes with 54XX[17] normal female sheep karyotype, comparable cell morphology, and growth curve to that of fresh tissue derived cells. Till date these cells have been passaged 21 times and show normal growth. These results suggest that normal, proliferative cells can be recovered from skin tissues stored at an elevated temperature of 35°C in livestock from 24-72 h of postmortem interval. Reprogramming of these cells to clone the dead animals or their use for cell therapies remain to be seen in future.

scholarly journals Evaluation of color stability of different temporary restorative materials

2015 ◽  
Vol 44 (5) ◽  
pp. 262-267 ◽  
Author(s):  
José Vitor Quinelli Mazaro ◽  
Luiz Miguel Minani ◽  
Adriana Cristina Zavanelli ◽  
Caroline Cantieri de Mello ◽  
Cleidiel Aparecido Araújo Lemos
Keyword(s):  
Storage Time ◽  
Color Change ◽  
Artificial Saliva ◽  
Color Stability ◽  
Time Interval ◽  
Acrylic Resins ◽  
Significant Difference ◽  
Restorative Materials ◽  
Before And After ◽  
Level Of Significance

AbstractIntroductionTemporary restorative materials are widely used, however, little is know about their color stability.Objectiveto evaluate the color stability of the following temporary restorative materials: acrylic and bis-acrylic resins after immersion in pigmenting solutions for different periods of storage.Material and methodFour materials were tested (Dêncor/Clássico, Protemp 4/3M ESPE; Structur 2 SC/Voco; Luxatemp AM Plus/DMG) and 30 test specimens (15 mm in diameter and 2 mm thick) per material were fabricated. They were divided according to the storage medium (artificial saliva, saliva + cola type soda, and saliva + coffee) and storage time intervals (2, 5, 7 and 15 days). Color measurements were made before and after immersions, with use of a spectrophotometer, by means of the CIE L*a*b* system. The data were analyzed by the analysis of variance and the Tukey Test, at a level of significance of 5%.ResultAcrylic resin presented greater color stability in comparison with bis-acrylic resins (p<0.001). When bis-acrylic resins were compared no significant difference was observed between the resins Structur and Luxatemp (p=0.767). As regards solutions tested, coffee showed the highest color change values (p<0.001), and the longer the storage time interval, the greater was the color change in all the temporary restorative materials analyzed (p<0.001).ConclusionAcrylic resin presented greater color stability in comparison with bis-acrylic resins (p<0.001). Coffee caused the greatest color change, and immersion time was determinant in color stability of the temporary materials analyzed.

scholarly journals The effects of Lactococcus lactis subsp. lactis and its supernatant on some bacteriological and sensory values in Rainbow trout (Onchorhynchus mykiss) fillets

2018 ◽  
Vol 9 (1) ◽  
Author(s):  
Asad Abbaspour Anbi ◽  
Vadood Razavilar ◽  
Moslem Neyriz Naghadehi ◽  
Masoud Seidgar ◽  
Ali Nekuiefard ◽  
...  
Keyword(s):  
Rainbow Trout ◽  
Lactococcus Lactis ◽  
Storage Time ◽  
Culture Medium ◽  
Live Cells ◽  
Human Consumption ◽  
Lactococcus Lactis Subsp ◽  
Live Bacteria ◽  
And Control ◽  
Control Samples

Lactic Acid Bacteria (LAB) have a great potential as bio-preservatives. The live cells and supernatant Lactococcus lactis subsp. lactis induced bacteriological changes in Onchorhynchus mykiss fillet by spray and immersion methods was studied during vacuum- packaged storage at 4 °C for 15 days. 40 kg of O. mykiss were prepared from a culture farm in Oshnavieh (Northwest Iran) and 112 fillet samples (100g) were prepared by aseptic method. L. lactis subsp. lactis (PTCC1336) bacteria was cultured in MRS culture medium. Its supernatant (2%, 4%) was extracted and 106 CFUml-1 dilutions of LAB were prepared and tested on the fillets to enhance their shelf life. All samples were evaluated regarding to growth of psychrotrophic, psychrophilic, mesophilic bacteria, molds and yeasts. Four characteristics including of odor, flavor, texture and color of fillets after and before cooking were evaluated for sensory analysis on days 1, 5, 10 and 15 and compared with control samples. The 4% supernatant and live bacteria were more effective than that of 2% and control (P<0.05). The amounts of corrosive bacteria in 4% and live cells in storage time were less than human consumption limits (7log CFUg-1), whereas in control and 2% supernatant treatments were more than that limits. The results showed that increasing the percentage of supernatant was more effective on bacteriologic factors and enhanced sensory characteristics of rainbow trout fillets (P<0.05).

scholarly journals Group-C Trisomy in Myeloid Metaplasia with Possible Leukemia

Blood ◽  
1964 ◽  
Vol 24 (6) ◽  
pp. 716-725 ◽  
Author(s):  
AVERY A. SANDBERG ◽  
TAKAAKI ISHIHARA ◽  
LOIS H. CROSSWHITE
Keyword(s):  
Bone Marrow ◽  
Blood Culture ◽  
Acute Leukemia ◽  
Chromosomal Abnormality ◽  
Blood Cells ◽  
Karyotype Analysis ◽  
Myeloproliferative Disorders ◽  
Normal Female ◽  
Myeloid Metaplasia ◽  
Definite Relationship

Abstract A chromosomal abnormality in marrow and blood cells has been found in only one patient out of a group of 20 subjects with myeloproliferative disorders other than leukemia. The abnormal karyotypic finding consisted of group C9 trisomy in a patient with myeloid metaplasia and an acute leukemia-like picture and indicates a definite relationship to acute leukemia. The latter has been shown to be not infrequently accompanied by C9 trisomy. The trisomy was accompanied by the presence of a substantial number of hypertetraploid cells in the marrow but not in the cultured blood cells. As a matter of fact, the blood culture yielded predominantly metaphases with 47 chromosomes (C9 trisomy) on the first examination and metaphases with 46 chromosomes and a normal female karyotype on the second occasion. The superiority of bone marrow karyotype analysis over that of blood cells in leukemic states is thus indicated.

scholarly journals Cribrilinid bryozoans from Pleistocene Mediterranean deep-waters, with the description of new species

2020 ◽  
pp. 1-23
Author(s):  
Antonietta Rosso ◽  
Emanuela Di Martino ◽  
Andrew N. Ostrovsky
Keyword(s):  
New Species ◽  
Deep Water ◽  
Time Interval ◽  
New Combinations ◽  
Mediterranean Species ◽  
Growth Morphology ◽  
Deep Waters ◽  
First Time

Abstract Cribrilinid bryozoans originating from Pleistocene deep-water sediments from two localities near Messina (Sicily, Italy)—Capo Milazzo (Gelasian) and Scoppo (Calabrian)—were examined. Five cribrilinid species were found, three in each locality and time interval, with only one species shared. Three species, Cribrilaria profunda n. sp., Glabrilaria transversocarinata n. sp., and Figularia spectabilis n. sp., are new to science. Of the two remaining species, Figularia figularis was already known from local fossil associations, whereas Glabrilaria pedunculata, a present-day Mediterranean species, is recorded for the first time as a fossil. New combinations are suggested for two species previously assigned to Puellina, Cribrilaria saldanhai (Harmelin, 2001) n. comb. and Cribrilaria mikelae (Harmelin, 2006) n. comb. The diagnosis of the genus Figularia was amended to include an erect growth morphology in addition to the encrusting form, and the occurrence of ooecia formed by the distal kenozooid. Following a literature revision of all species currently assigned to Figularia, the new combinations Vitrimurella capitifera (Canu and Bassler, 1929) n. comb. and Hayamiellina quaylei (Powell, 1967a) n. comb. are suggested, and problematic species are listed and briefly discussed. UUID: http://zoobank.org/b7b36152-bf7b-4e00-b6ec-2614b2a58f1b

scholarly journals Production of Kunapajala and Sanjibani, Their Nutritional Contributions, Microbial and Pesticidal Effect

2019 ◽  
pp. 1-11
Author(s):  
Bishal Chakraborty ◽  
Indrajit Sarkar ◽  
Swathi Kulukunde ◽  
Soumen Maitra ◽  
Arpita Mandal Khan ◽  
...  
Keyword(s):  
Storage Time ◽  
West Bengal ◽  
Nutrient Content ◽  
Microbial Load ◽  
Soil Science ◽  
Time Interval ◽  
Chemical Fertilizers ◽  
Agricultural Chemistry ◽  
Microbial Analysis ◽  
Maximum Utilization

A study on nutritional and microbial analysis of Kunapajala with different storage time interval was conducted in the Department of Soil Science & Agricultural Chemistry and the Department of Plant Pathology, UBKV, Coochbehar-736165, West Bengal during March, 2019. The motive of this work was to estimate the physicochemical properties, macro and micro nutrient content and various microbial load of Kunapajala with different storage time interval. Kunapajala had the highest P, K, Ca, Mg, Fe, Zn, Cu & Mn 40 days after preparation and it had highest N and S 20 days after preparation. It had the highest beneficial microbial load of Fungi, Actinomycetes, Pseudomonus, Phosphorus Solubilising Bacteria (PSB), Azotobacter, Azospirillum, Rhizobium and Trichoderma 40 days after preparation. So, continuous foliar and soil application of Kunapajala from 20 days after preparation to 40 days after preparation was beneficial to get maximum utilization. Moreover, Kunapajala can be used as an alternative against chemical fertilizers and pesticides to develop organic farming.

Microbial Growth and Successions on Steaks as Influenced by Packaging Procedures1

1989 ◽  
Vol 52 (4) ◽  
pp. 236-239 ◽  
Author(s):  
HAMDI A. AHMAD ◽  
JOHN A. MARCHELLO
Keyword(s):  
Storage Time ◽  
Microbial Growth ◽  
Growth Patterns ◽  
Similar Growth

The influence of retail packaging procedures: (1) Borden's Resinite film overwrap; (2) Gas (1% CO, 40% O2, 59% N2) flushed for 2 min then overwrapped with the Resinite film; (3) packaged in a barrier bag and sealed with no evacuation of air on microbial growth and succession on steak surfaces was studied during 12 d of storage at 4°C. Growth on top and bottom surfaces for all packaged steaks did not differ significantly within most sampling periods. Similar growth patterns were observed on both steak surfaces, increasing (P &lt;0.05) between d 3, 6, and 9 of storage. Pseudomonas dominated the microflora on steaks packaged in all treatments. However, Pseudomonas was the dominant organism on the steaks packaged in treatment 3 only through d 6; thereafter the microflora were dominated by Serratia. The numbers of Micrococcus, Brochothrix, and coryneforms were also increased with storage time in all treatments.

scholarly journals PBMC Fixation and Processing for Chromium Single-Cell RNA Sequencing

2018 ◽  
Author(s):  
Jinguo Chen ◽  
Foo Cheung ◽  
Rongye Shi ◽  
Huizhi Zhou ◽  
Wenrui Wenrui ◽  
...  
Keyword(s):  
Single Cell ◽  
Rna Sequencing ◽  
Mononuclear Cells ◽  
Human Peripheral Blood ◽  
Live Cells ◽  
Similar Proportion ◽  
Marker Genes ◽  
Primary Cells ◽  
Single Cell Rna Sequencing ◽  
Cell Fixation

AbstractBackgroundInterest in single-cell transcriptomic analysis is growing rapidly, especially for profiling rare or heterogeneous populations of cells. In almost all reported works investigators have used live cells, which introduces cell stress during preparation and hinders complex study designs. Recent studies have indicated that cells fixed by denaturing fixative can be used in single-cell sequencing, however they did not usually work with most types of primary cells including immune cells.MethodsThe methanol-fixation and new processing method was introduced to preserve human peripheral blood mononuclear cells (PBMCs) for single-cell RNA sequencing (scRNA-Seq) analysis on 10X Chromium platform.ResultsWhen methanol fixation protocol was broken up into three steps: fixation, storage and rehydration, we found that PBMC RNA was degraded during rehydration with PBS, not at cell fixation and up to three-month storage steps. Resuspension but not rehydration in 3X saline sodium citrate (SSC) buffer instead of PBS preserved PBMC RNA integrity and prevented RNA leakage. Diluted SSC buffer did not interfere with full-length cDNA synthesis. The methanol-fixed PBMCs resuspended in 3X SSC were successfully implemented into 10X Chromium standard scRNA-seq workflows with no elevated low quality cells and cell doublets. The fixation process did not alter the single-cell transcriptional profiles and gene expression levels. Major subpopulations classified by marker genes could be identified in fixed PBMCs at a similar proportion as in live PBMCs. This new fixation processing protocol also worked in several other fixed primary cell types and cell lines as in live ones.ConclusionsWe expect that the methanol-based cell fixation procedure presented here will allow better and more effective batching schemes for a complex single cell experimental design with primary cells or tissues.

scholarly journals Improving IBD diagnosis and monitoring by understanding preanalytical, analytical and biological fecal calprotectin variability

2018 ◽  
Vol 56 (11) ◽  
pp. 1926-1935 ◽  
Author(s):  
Andrea Padoan ◽  
Renata D’Incà ◽  
Maria Luisa Scapellato ◽  
Rudi De Bastiani ◽  
Roberta Caccaro ◽  
...  
Keyword(s):  
Room Temperature ◽  
Storage Time ◽  
Storage Temperature ◽  
Clinical Effectiveness ◽  
Fecal Calprotectin ◽  
Time Interval ◽  
Biological Variability ◽  
Reference Change Value ◽  
The Difference ◽  
Clinically Significant

Abstract Background: The appropriate clinical use of fecal calprotectin (fCal) might be compromised by incomplete harmonization between assays and within- and between-subjects variability. Our aim was to investigate the analytical and biological variability of fCal in order to provide tools for interpreting fCal in the clinical setting. Methods: Experiments were conducted to investigate the effects of temperature and storage time on fCal. Thirty-nine controls were enrolled to verify biological variability, and a case-control study was conducted on 134 controls and 110 IBD patients to compare the clinical effectiveness of three different fCal assays: ELISA, CLIA and turbidimetry. Results: A 12% decline in fCal levels was observed within 24 h following stool collection irrespective of storage temperature. Samples were unstable following a longer storage time interval at room temperature. Within- and between-subjects fCal biological variability, at 31% and 72% respectively, resulted in a reference change value (RCV) in the region of 100%. fCal sensitivity in distinguishing between controls and IBD patients is satisfactory (68%), and the specificity high (93%) among young (<65 years), but not among older (≥65 years) subjects (ROC area: 0.584; 95% CI: 0.399–0.769). Among the young, assays have different optimal thresholds (120 μg/g for ELISA, 50 μg/g for CLIA and 100 μg/g for turbidimetry). Conclusions: We recommend a standardized preanalytical protocol for fCal, avoiding storage at room temperature for more than 24 h. Different cutoffs are recommended for different fCal assays. In monitoring, the difference between two consecutive measurements appears clinically significant when higher than 100%, the fCal biological variability-derived RCV.

scholarly journals Ovarian Gonadoblastoma with the Karyotype of 46, XX: A Case Report

2021 ◽  
Author(s):  
Fengju Zhao ◽  
Yingchao Zhao ◽  
Biyuan Xing ◽  
Zhao Liu
Keyword(s):  
Y Chromosome ◽  
Karyotype Analysis ◽  
Pelvic Mass ◽  
Peripheral Blood Lymphocytes ◽  
Normal Female ◽  
Pathological Analysis ◽  
Forkhead Box ◽  
Routine Physical Examination ◽  
Foxo Signaling Pathway ◽  
Chromosome Material

Abstract Gonadoblastoma is a rare tumor comprised of sex cord derivatives and germ cells. The risk for developing gonadoblastoma increases significantly in patients who possess a Y chromosome or Y chromosome material. A 49-year-old Chinese woman found a pelvic mass during a routine physical examination. Pathological analysis after surgery indicated that the tumor was unilateral ovarian gonadoblastoma with dysgerminoma. Compared with other cases in the literature, our patient was the oldest, and the tumor mass was smaller. Karyotype analysis of peripheral blood lymphocytes revealed that the woman had a 46, XX female karyotype. Whole-exon sequencing revealed that some mutations, such as altered somatic genes in the Forkhead box protein O (FoxO) signaling pathway and KIT, might cause the disease. In conclusion, we described a rare case of gonadoblastoma in a woman who had normal routine menstruation, sexual development, and successful pregnancies and possessed a normal female 46, XX karyotype.

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