scholarly journals An in vitro feasibility investigation considering primary human melanocytes for spray- grafting of freshly isolated autologous skin cells for burn treatment and a clinical case report

2019 ◽  
pp. 1-8
Author(s):  
Jörg C. Gerlach ◽  
C. Johnen ◽  
B. Hartmann ◽  
J. Plettig ◽  
K. Bräutigam ◽  
...  
Keyword(s):  
Cell Morphology ◽  
Skin Color ◽  
In Vitro Model ◽  
Primary Cultures ◽  
Cell Suspensions ◽  
Epidermal Keratinocytes ◽  
Cell Counts ◽  
Skin Coloration ◽  
Vitro Model

A skin cell-spray grafting technique that enables the on-site application of freshly isolated autologous single cell suspensions was already applied in many cases on caucasian patients with low skin coloration. Our project hypothesis is that these suspensions contain keratinocytes and vital melanocytes, that are of particular interest for the treatment of patients of darker skin color. To test this, we applied an in vitro model, wherein the feasibility of i) isolating and ii) spraying of freshly isolated autologous melanocyte-keratinocyte cell suspensions was investigated. Primary human epidermal keratinocytes (HEKs) and melanocytes (MCs) were isolated from skin biopsies (n=8). Biochemical parameter, cell counts, cell morphology, growth behavior and immunofluorescence results were compared in two groups using MC cultures and co-cultures of MCs with HEKs. Case information on using the method clinically with one patient is included. The sprayed mixed cell suspensions proliferated in all groups without measurable loss of viability, and MCs exhibited a regular cell morphology in monoculture up to passage 4°. The sprayed MCs and HEKs demonstrated in vitro glucose and lactate metabolism that was comparable to the pipetted controls. In co-culture, well distributed CK14+ HEKs and NKI/beteb+ MCs could be demonstrated, which interacted in the in vitro model. The ratio of HEKs : MCs in our primary cultures were microscopically counted (n=8 each) as mean +/- SD 1,211,000 (+/- 574,343) HEK : 99,625 (+/- 59,025) MC; i.e., a ratio of approx. 12 : 1. Using the isolation method clinically for a patient with dark skin coloration after suffering severe second-degree burns shows a satisfying re-pigmentation of the resulting wound post healing. Freshly isolated spray-on melanocyte/keratinocyte suspensions provide for a considerable amount of viable HEKs and MCs. Using MCs in spray-grafting suspensions could represent a promising approach for treating severe partial-thickness burns and innovative therapy developments that also aim to address cosmetic aspects.

An in vitro model for the study of human parathyroid gland tissue: Single cell suspensions and monolayer cultures

1982 ◽  
Vol 181 (2) ◽  
pp. 147-154 ◽  
Author(s):  
P. K. Wagner ◽  
A. Knuth ◽  
U. Krause ◽  
H. Gabbert ◽  
Th. Schärfe ◽  
...  
Keyword(s):  
Single Cell ◽  
Parathyroid Gland ◽  
In Vitro Model ◽  
Cell Suspensions ◽  
Monolayer Cultures ◽  
Gland Tissue ◽  
Vitro Model

scholarly journals Atlantic salmon cardiac primary cultures: An in vitro model to study viral host pathogen interactions and pathogenesis

PLoS ONE ◽  
2017 ◽  
Vol 12 (7) ◽  
pp. e0181058 ◽  
Author(s):  
Patricia A. Noguera ◽  
Bianka Grunow ◽  
Matthias Klinger ◽  
Katherine Lester ◽  
Bertrand Collet ◽  
...  
Keyword(s):  
Atlantic Salmon ◽  
In Vitro Model ◽  
Primary Cultures ◽  
Host Pathogen Interactions ◽  
Host Pathogen ◽  
Vitro Model

Renal cell carcinoma primary cultures as in vitro model to study genomic profile of parental tumor tissues

2008 ◽  
Vol 6 (9) ◽  
pp. 117
Author(s):  
I. Cifola ◽  
C. Bianchi ◽  
E. Mangano ◽  
S. Bombelli ◽  
S. Ferrero ◽  
...  
Keyword(s):  
Renal Cell Carcinoma ◽  
Cell Carcinoma ◽  
Renal Cell ◽  
In Vitro Model ◽  
Primary Cultures ◽  
Genomic Profile ◽  
Tumor Tissues ◽  
Parental Tumor ◽  
Vitro Model

scholarly journals Anti-Photoaging Effect of Plant Extract Fermented with Lactobacillus buchneri on CCD-986sk Fibroblasts and HaCaT Keratinocytes

2020 ◽  
Vol 11 (1) ◽  
pp. 3 ◽  
Author(s):  
Yun-Mi Kang ◽  
Chul-Hee Hong ◽  
Sa-Haeng Kang ◽  
Dong-Seok Seo ◽  
Seong-Oh Kim ◽  
...  
Keyword(s):  
In Vitro Model ◽  
Type I Collagen ◽  
Dermal Fibroblasts ◽  
Epidermal Keratinocytes ◽  
Type I ◽  
Mrna Levels ◽  
Protective Effects ◽  
Lactobacillus Buchneri ◽  
Vitro Model

Ultraviolet (UV) exposure triggers the abnormal production of reactive oxygen (ROS) species and the expression of matrix metalloproteinases (MMPs) that are responsible for photoaging. Probiotics are widely used in healthcare and for immune enhancement. One probiotic, Lactobacillus buchneri is found in Kimchi. This study was aimed at assessing the anti-photoaging effect of plant extracts fermented with L. buchneri (PELB) to develop functional cosmetics. We investigated the anti-photoaging effect of PELB in a UVB-induced photoaging in vitro model and selected effective extracts using the elastase inhibition assay, ELISA for Type I procollagen and collagenase-1, and quantitative real time PCR. Normal human dermal fibroblasts and epidermal keratinocytes were pre-treated with PELB and exposed to UVB. We found that PELB decreased elastase activity and increased type I collagen expression in a UVB-induced photoaging in vitro model. In addition, PELB greatly reduced collagenase activity and MMP mRNA levels in a UVB-induced photoaging in vitro model. Furthermore, PELB promoted the expression of moisture factor and anti-oxidant enzymes in a UVB-induced photoaging in vitro model. These results indicated that the PELB could be potential candidates for the protective effects against UVB-induced photoaging. Overall, these results suggest that PELB might be useful natural components of cosmetic products.

scholarly journals Ovine fetuses from slaughterhouses: A useful source for neural cell primary cultures

2021 ◽  
Author(s):  
Vittorio Farina ◽  
Sergio Domenico Gadau ◽  
Gianluca Lepore ◽  
Marcella Carcupino ◽  
Marco Zedda
Keyword(s):  
In Vitro Model ◽  
Primary Cultures ◽  
Medical Procedures ◽  
Cell Type ◽  
Experimental Animals ◽  
Vitro Model ◽  
Set Up ◽  
Very High ◽  
Β Tubulin

A lot of evidence demonstrates that sheep could represent an experimental model to set up medical procedures in view of their application on humans. Sheep are chosen as models for human biomechanical studies because their skeleton has some similarities to humans. The aim of this work was to set up sheep primary cultures from ovine fetuses at different ages, from pregnant uteri retrieved at local abattoirs. Cell characterization showed that one cell population was immunopositive to GFAP and identifiable as astrocytes, whereas a second cell type was III β-tubulin-positive, and hence classified as neurons. At 60-day old fetus is suitable to obtain neurons, whereas in a 90-day old fetus the cell culture is predominantly characterized by glial cells. The procedure here proposed is inexpensive, in fact, collecting fetuses during sheep slaughtering is a cost-saving option, unlike common experimental animals such as mice, rats, rabbits, that require very high economical efforts. Finally, our protocol fully eliminates the need of animal killing, being living animals replaced by a validated in vitro model in agreement with the 3Rs statement.

scholarly journals Effect of bacterial lipopolysaccharide in primary cultures of human gingival fibroblasts

2017 ◽  
Vol 14 (2) ◽  
Author(s):  
Maria Cecilia Verutti ◽  
Octavio González ◽  
John González ◽  
Gloria Moreno
Keyword(s):  
Healthy Donor ◽  
In Vitro Model ◽  
Periodontal Diseases ◽  
Primary Cultures ◽  
Cell Number ◽  
Human Gingival Fibroblasts ◽  
Gingival Fibroblasts ◽  
Short Period ◽  
Vitro Model

Summary: Introduction: different factors participate in the pathogenesis ofperiodontal diseases. One factor is the interaction between the fibroblasts and derived productsfrom the microorganisms found in the periodontal environment. Objective: In this work, cultures ofhuman gingival fibroblasts from a healthy donor were used to characterize the in vitro responses tobacterial lipopolysaccharide. Methods: the proliferative response was evaluated using cell count andexpression of CD14 was assessed by flow cytometry. Results: after 24 hours of culture an increase inthe cell number was detected in cultures treated with 1.0 mg/mL LPS, but these differences were notstatistically significant. Human gingival fibroblasts express CD14, but its expression decreases incells cultivated after a short period of time. Nevertheless, lipopolysaccharide helps to recover theexpression of CD14 in fibroblast after 24 hours of culture. Conclusion: preliminary result suggeststhat expression of CD14 on gingival fibroblasts could be modulated by this bacterial derived toxin.The primary culture of human gingival fibroblasts allows the establishment of an in vitro model toevaluate different processes in development of the periodontal diseases. Key words: Gingivalfibroblasts. Lipopolysaccharide. Periodontal disease.

An In Vitro Model of Differentiated Human Airway Epithelia: Methods for Establishing Primary Cultures

2003 ◽  
pp. 115-137 ◽  
Author(s):  
Philip H. Karp ◽  
Thomas O. Moninger ◽  
S. Pary Weber ◽  
Tamara S. Nesselhauf ◽  
Janice L. Launspach ◽  
...  
Keyword(s):  
In Vitro Model ◽  
Primary Cultures ◽  
Airway Epithelia ◽  
Human Airway ◽  
Vitro Model ◽  
Human Airway Epithelia

scholarly journals Effect of Carbohydrate Composition in Barley and Oat Cultivars on Microbial Ecophysiology and Proliferation of Salmonella enterica in an In Vitro Model of the Porcine Gastrointestinal Tract

2009 ◽  
Vol 75 (22) ◽  
pp. 7006-7016 ◽  
Author(s):  
Robert Pieper ◽  
J�r�me Bindelle ◽  
Brian Rossnagel ◽  
Andrew Van Kessel ◽  
Pascal Leterme
Keyword(s):  
Gastrointestinal Tract ◽  
Salmonella Enterica ◽  
In Vitro Model ◽  
Relative Proportion ◽  
Canonical Ordination ◽  
Barley Cultivars ◽  
Cell Counts ◽  
Oat Cultivars ◽  
Vitro Model

ABSTRACT The influence of the carbohydrate (CHO) composition of cereal cultivars on microbial ecophysiology was studied using an in vitro model of the porcine gastrointestinal tract. Ten hull-less barley cultivars, six barley cultivars with hulls, six oat cultivars, and six oat groats that differed in β-glucan, nonstarch polysaccharide (NSP), and starch contents and starch type were hydrolyzed enzymatically and incubated for 72 h with pig feces. Fermentation kinetics were modeled, and microbial compositions and short-chain fatty acid (SCFA) profiles were analyzed using terminal restriction fragment length polymorphism and gas chromatography. Cluster analysis and canonical ordination revealed different effects on fermentation and microbial ecology depending on the type of CHO and cultivar. First, in cultivars of barley with hulls and oats, the cellulose and insoluble NSP contents (i) increased Ruminococcus flavefaciens-like and Clostridium xylanolyticum-like phylotypes, (ii) increased acetate production, and (iii) decreased fermentation activity. Second, in hull-less barley cultivars the β-glucan, amylose, amylopectin, crude protein, and soluble NSP contents determined the microbial community composition and activity as follows: (i) the amylose contents of the hull-less barley varieties increased the butyrate production and the abundance of Clostridium butyricum-like phylotypes, (ii) the β-glucan content determined the total amounts of SCFA, and (iii) the amylopectin and starch contents affected the abundance of Clostridium ramosum-like phylotypes, members of Clostridium cluster XIVa, and Bacteroides-like bacteria. Finally, the effect of CHO on proliferation of Salmonella enterica in the model was determined. Salmonella cell counts were not affected, but the relative proportion of Salmonella decreased with hull-less barley cultivars and increased with oat cultivars as revealed by quantitative PCR. Our results shed light on the complex interactions of cereal CHO with intestinal bacterial ecophysiology and the possible impact on host health.

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