scholarly journals In vitro and ex vivo Functional Evaluation of a Hollow Fiber-type Bioartificial Liver Module Containing ES Cell-derived Hepatocyte-like Cells

2018 ◽  
Vol 7 (0) ◽  
pp. 18-27 ◽  
Author(s):  
Hiroshi Mizumoto ◽  
Naoki Amimoto ◽  
Toru Miyazawa ◽  
Hideki Tani ◽  
Kaoru Ikeda ◽  
...  
Keyword(s):  
Hollow Fiber ◽  
Ex Vivo ◽  
Fiber Type ◽  
Bioartificial Liver ◽  
Functional Evaluation ◽  
Es Cell

Evaluation of the mass transfer rate using computer simulation in a three-dimensional interwoven hollow fiber-type bioartificial liver

2018 ◽  
Vol 40 (11-12) ◽  
pp. 1567-1578 ◽  
Author(s):  
Ryoichi Sakiyama ◽  
Hiroyuki Hamada ◽  
Brandon Blau ◽  
Nora Freyer ◽  
Katrin Zeilinger ◽  
...  
Keyword(s):  
Mass Transfer ◽  
Computer Simulation ◽  
Hollow Fiber ◽  
Transfer Rate ◽  
Fiber Type ◽  
Mass Transfer Rate ◽  
Three Dimensional ◽  
Bioartificial Liver

Perfusion Circuit Concepts for Hollow-Fiber Bioreactors Used as in Vitro Cell Production Systems or Ex Vivo Bioartificial Organs

2011 ◽  
Vol 34 (5) ◽  
pp. 410-421 ◽  
Author(s):  
Stephen C. Balmert ◽  
Daniel McKeel ◽  
Fabio Triolo ◽  
Bruno Gridelli ◽  
Katrin Zeilinger ◽  
...  
Keyword(s):  
Hollow Fiber ◽  
Production Systems ◽  
Ex Vivo ◽  
Cell Production ◽  
Bioartificial Organs ◽  
Hollow Fiber Bioreactors

scholarly journals Comparative and Functional Evaluation of In Vitro Generated to Ex Vivo CD8 T Cells

2012 ◽  
Vol 189 (7) ◽  
pp. 3411-3420 ◽  
Author(s):  
Džana D. Dervović ◽  
Maria Ciofani ◽  
Korosh Kianizad ◽  
Juan Carlos Zúñiga-Pflücker
Keyword(s):  
T Cells ◽  
Cd8 T Cells ◽  
Ex Vivo ◽  
Functional Evaluation

Neonatal Satellite Cells Form Small Myotubes In Vitro

2016 ◽  
Vol 96 (3) ◽  
pp. 331-338 ◽  
Author(s):  
P.L. Carvajal Monroy ◽  
S. Grefte ◽  
A.M. Kuijpers-Jagtman ◽  
J.W. Von den Hoff ◽  
F.A.D.T.G. Wagener
Keyword(s):  
Cleft Palate ◽  
Satellite Cells ◽  
Ex Vivo ◽  
Fiber Type ◽  
Careful Consideration ◽  
Speech Development ◽  
Proliferation Index ◽  
Delayed Differentiation ◽  
Fusion Index

Although palatal muscle reconstruction in patients with cleft palate takes place during early childhood, normal speech development is often not achieved. We hypothesized that the intrinsic properties of head satellite cells (SCs) and the young age of these patients contribute to the poor muscle regeneration after surgery. First, we studied the fiber type distribution and the expression of SC markers in ex vivo muscle tissue from head (branchiomeric) and limb (somite-derived) muscles from neonatal (2-wk-old) and young (9-wk-old) rats. Next, we cultured SCs isolated from these muscles for 5, 7, and 9 d, and investigated the in vitro expression of SC markers, as well as changes in proliferation, early differentiation, and fusion index (myotube formation) in these cells. In our ex vivo samples, we found that virtually all myofibers in both the masseter (Mass) and the levator veli palatini (LVP) muscles contained fast myosin heavy chain (MyHC), and a small percentage of digastric (Dig) and extensor digitorum longus myofibers also contained slow MyHC. This was independent of age. More SCs were found in muscles from neonatal rats as compared with young rats [17.6 (3.8%) v. 2.3 (1.6%); P < 0.0001]. In vitro, young branchiomeric head muscle (BrHM) SCs proliferated longer and differentiated later than limb muscle SCs. No differences were found between SC cultures from the different BrHMs. SC cultures from neonatal muscles showed a much higher proliferation index than those from young animals at 5 d (0.8 v. 0.2; P < 0.001). In contrast, the fusion index in neonate SCs was about twice as low as that in SCs from young muscles at 9 d [27.6 (1.4) v. 62.8 (10.2), P < 0.0001]. In conclusion, SCs from BrHM differ from limb muscles especially in their delayed differentiation. SCs from neonatal muscles form myotubes less efficiently than those from young muscles. These age-dependent differences in stem cell properties urge careful consideration for future clinical applications in patients with cleft palate.

oct-4 Is Essential for Aneuploidy/Polyploidy-Tolerance in Embryonic Stem (ES) Cells and the Fate of Tetraploid ES Cells during Early Differentiation.

Blood ◽  
2005 ◽  
Vol 106 (11) ◽  
pp. 311-311
Author(s):  
Charlie R. Mantel ◽  
Ying Guo ◽  
Myung-Kwan Han ◽  
Sara Roharbaugh ◽  
Barbara Graham-Evans ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Cell Cultures ◽  
Chromosomal Aberrations ◽  
Ex Vivo ◽  
Es Cells ◽  
Somatic Cells ◽  
Es Cell ◽  
Present Evidence ◽  
Mitotic Checkpoints

Abstract Maintenance of a euploid ES cell genome during ex-vivo expansion and differentiation to hematopoietic cells is essential for their safe use in therapeutic intervention and regenerative medicine. It is estimated that about half of all spontaneous mutations that occur in cultured murine ES cells are attributable to chromosomal non-disjunction and chromosome loss. This type of mutation is essentially undetectable in somatic cell cultures of comparable culture age. The mechanisms responsible for this difference between ES and somatic cells are unknown, but are likely related to the absence of a rigorous G1 cell cycle checkpoint and other peculiarities in ES cell cycle regulation compared to somatic cells. The fate of aneuploid ES cells during differentiation has not been systematically studied. Here, we report a remarkable tolerance for aneuploidy/polyploidy in murine ES cells in-vitro when challenged by mitotic stress that is induced by failed cell division followed by mitotic slippage resulting in stable, cycling, tetraploid/polyploid ES cell lines (oscillating between 4N and 8N). We present evidence supporting the idea that canonical apoptosis is uncoupled from mitotic checkpoints in undifferentiated ES cells in contrast to somatic cells and embryoid body (EB) cells. We also present evidence suggesting similar behavior occurs in human ES cell cultures. Uncoupling was associated with low levels of the pro-apoptotic form of phospho-BAD (p-ser128) combined with high expression of anti-apoptotic Survivin in ES compared to EB cells. This suggests uncoupling from mitotic checkpoints may be related to a heightened apoptotic threshold in ES cells compared to EB/somatic cells. Interestingly, culture of ES cells under hypoxic conditions also generated polyploid cells. The polyploid ES cells did not appear to be trophoblastic cells because they continue to express SSEA-1 and other markers of pluripotency. However, we demonstrate that (re-)coupling occurs very early in the differentiation process because only diploid ES cells contribute to EB formation while tetraploid cells do not when EBs are generated from tetraploid/diploid-mosaic cultures. This is, to our knowledge, the first evidence that there may be a potent in-vitro barrier to cells with numerical chromosomal aberrations from contributing to differentiated cells to be used in therapeutic settings. Finally, using a conditional oct-4 knock-down ES cell line, we demonstrate that the self-renewal regulating transcription factor, oct-4, is essential for maintaining the aneuploidy/polyploidy tolerant state. We conclude that aneuploidy/polyploidy-tolerance in pluripotent ES cells in-vitro is an expected occurrence when viewed within the context of ex-vivo ES cell culture where apoptotic culling of cells with chromosomal aberrations, which normally occurs in vertebrate embryos during the peri-implantation period in-vivo, has been artificially interrupted by ES cell derivation. This behavior likely contributes significantly to spontaneous aneuploidization in murine and human ES cell cultures. These data not only lend insight into mechanisms of aneuploidy in ES cell cultures but may also have implications for mechanisms of aneuploidy during tumorigenesis.

scholarly journals Rapid and efficient generation of antigen-specific isogenic T cells from cryopreserved blood samples

2021 ◽  
Author(s):  
Anneke Eerkens ◽  
Annege Vledder ◽  
Nienke van Rooij ◽  
Floris Foijer ◽  
Hans W Nijman ◽  
...  
Keyword(s):  
T Cells ◽  
T Cell ◽  
T Cell Activation ◽  
Cell Activation ◽  
Gene Editing ◽  
Ex Vivo ◽  
Immune Monitoring ◽  
Functional Evaluation ◽  
Starting Point

Objectives: CRISPR/Cas9-mediated gene editing has been leveraged for the modification of human and mouse T cells. However, limited experience is available on the application of CRISPR/Cas9 electroporation in cryopreserved T cells collected during e.g. clinical trials. Methods: PBMCs from healthy donors were used to generate knockout T cell models for interferon-gamma (IFNg), Cbl Proto-Oncogene B (CBLB), Fas cell surface death receptor (Fas) and T cell receptor (TCR alpha and beta) genes. The effect of CRISPR-cas9-mediated gene editing on T cells was evaluated using apoptosis assays, cytokine bead arrays and ex vivo and in vitro stimulation assays. Results: Our results demonstrate that CRISPR/Cas9-mediated gene editing of ex vivo T cells is efficient and does not overtly affect T cell viability. Cytokine release and T cell proliferation were not affected in gene edited T cells. Interestingly, memory T cells were more susceptible to CRISPR/Cas9 gene editing than naive T cells. Ex vivo and in vitro stimulation with antigens resulted in equivalent antigen-specific T cell responses in gene-edited and untouched control cells; making CRISPR/Cas9-mediated gene editing compatible with clinical antigen-specific T cell activation and expansion assays. Conclusion: Here, we report an optimized protocol for rapid, viable and highly efficient genetic modification in ex vivo human antigen specific T cells, for subsequent functional evaluation and/or expansion. Our platform extends CRISPR/Cas9-mediated gene editing for use in gold-standard clinically-used immune-monitoring pipelines and serves as a starting point for development of analogous approaches such as those including transcriptional activators and or epigenetic modifiers.

Effects of the Tibetan herbal preparation PADMA 28 on blood lipids and lipid oxidisability in subjects with mild hypercholesterolaemia

VASA ◽  
2005 ◽  
Vol 34 (1) ◽  
pp. 11-17 ◽  
Author(s):  
Brunner-La Rocca ◽  
Schindler ◽  
Schlumpf ◽  
Saller ◽  
Suter
Keyword(s):  
Endothelial Dysfunction ◽  
Blood Lipids ◽  
Ex Vivo ◽  
Hdl Cholesterol ◽  
Blood Lipid ◽  
Tibetan Medicine ◽  
Double Blind ◽  
Intraarterial Infusion ◽  
Padma 28

Background: Previous studies showed an anti-atherosclerotic effect of PADMA 28, an herbal formula based on Tibetan medicine. As the mechanisms of action are not fully understood, we investigated whether PADMA 28 may lower blood lipids and lipid oxidisability, and affect early endothelial dysfunction. Patients and methods: Sixty otherwise healthy subjects with total cholesterol ≥5.2 mmol/l and < 8.0 mmol/l were randomly assigned to placebo or PADMA 28, 3 x 2 capsules daily, for 4 weeks (double-blind). Blood lipids (total, LDL-, and HDL-cholesterol, triglycerides, Apo-lipoprotein A1 and B) and ex vivo lipid oxidisability were measured before and after treatment. In a subset of 24 subjects, endothelial function was assessed using venous occlusion plethysmography with intraarterial infusion of acetylcholine. Isolated LDL and plasma both untreated and pre-treated with PADMA 28 extract were oxidised by the radical generator AAPH. Conjugated diene formation was measured at 245 nm. Results: Blood lipids did not change during the study in both groups. In contrast to previous reports in mild hypercholesterolaemia, no endothelial dysfunction was seen and, consequently, was not influenced by therapy. Ex vivo blood lipid oxidisability was significantly reduced with PADMA 28 (area under curve: 5.29 ± 1.62 to 4.99 ± 1.46, p = 0.01), and remained unchanged in the placebo group (5.33 ± 1.88 to 5.18 ± 1.78, p > 0.1). This effect persisted one week after cessation of medication. In vitro experiments confirmed the prevention of lipid peroxidation in the presence of PADMA 28 extracts. Persistent protection was also seen for LDL isolated from PADMA 28-pretreated blood after being subjected to rigorous purification. Conclusions: This study suggests that the inhibition of blood lipid oxidisability by PADMA 28 may play a role in its anti-atherosclerotic effect.

The Role of Polyphenols in the Modulation of Plasma Non-Enzymatic Antioxidant Capacity (NEAC)

2012 ◽  
Vol 82 (3) ◽  
pp. 228-232 ◽  
Author(s):  
Mauro Serafini ◽  
Giuseppa Morabito
Keyword(s):  
Antioxidant Capacity ◽  
Dietary Intervention ◽  
Ex Vivo ◽  
The Body ◽  
Dietary Polyphenols ◽  
Enzymatic Antioxidant ◽  
Endogenous Antioxidant

Dietary polyphenols have been shown to scavenge free radicals, modulating cellular redox transcription factors in different in vitro and ex vivo models. Dietary intervention studies have shown that consumption of plant foods modulates plasma Non-Enzymatic Antioxidant Capacity (NEAC), a biomarker of the endogenous antioxidant network, in human subjects. However, the identification of the molecules responsible for this effect are yet to be obtained and evidences of an antioxidant in vivo action of polyphenols are conflicting. There is a clear discrepancy between polyphenols (PP) concentration in body fluids and the extent of increase of plasma NEAC. The low degree of absorption and the extensive metabolism of PP within the body have raised questions about their contribution to the endogenous antioxidant network. This work will discuss the role of polyphenols from galenic preparation, food extracts, and selected dietary sources as modulators of plasma NEAC in humans.

In-vitro/ex-vivo HIPEC-Modelle zur Therapieeffizienzprüfung

2013 ◽  
Vol 51 (08) ◽  
Author(s):  
C Ulmer ◽  
L Schaaf ◽  
W Zopf ◽  
W Steurer
Keyword(s):  
Ex Vivo
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