Improving Quality of In vitro Methods: A GIVIMP Certification Program

SummaryThe activity of several synthetic compounds, rated from good to poor (or inactive) fibrinolytic activators, has been assessed by two different commonly-used in vitro methods. Compounds shown to be active over a narrow concentration range in the hanging clot test were shown to be inhibitors of plasmin and trypsin in the casein-olytic test. The inhibitory activity of these compounds was shown to increase with increasing substrate concentration and apparent activity in the hanging clot test. Possible explanations and relevance of these observations are discussed.

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No differences in development/quality of embryos cultured in sequential and single-step medium regarding of female age and method of in vitro fertilization: prospective study

10.26226/morressier.573c1511d462b80296c9842e ◽

2016 ◽

Author(s):

Liljana Bacer Kermavner

Keyword(s):

In Vitro Fertilization ◽

Prospective Study ◽

Single Step ◽

Female Age ◽

Vitro Fertilization

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THE EFFECT OF EPIDERMAL GROWTH FACTOR ON QUALITY OF BOVINE OOCYTES AGED IN VITRO

Genetika i razvedenie zhivotnyh ◽

10.31043/2410-2733-2018-4-29-33 ◽

2018 ◽

pp. 29-33

Author(s):

G. SINGINA ◽

◽

E. SHEDOVA ◽

I. LEBEDEVA ◽

◽

...

Keyword(s):

Growth Factor ◽

Epidermal Growth Factor ◽

Epidermal Growth ◽

Bovine Oocytes

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Bioactive Peptides Sensitize Cells to Anticancer Effects of Oxaliplatin in Human Colorectal Cancer Xenografts in Nude Mice

Protein and Peptide Letters ◽

10.2174/0929866526666190405124955 ◽

2019 ◽

Vol 26 (7) ◽

pp. 512-522

Author(s):

Xian Li ◽

Long Xia ◽

Xiaohui Ouyang ◽

Qimuge Suyila ◽

Liya Su ◽

...

Keyword(s):

Quality Of Life ◽

Colorectal Cancer ◽

Nude Mice ◽

Low Dose ◽

Bioactive Peptides ◽

Human Colorectal Cancer ◽

Short Term ◽

Hct 116

Background: Despite new agent development and short-term benefits in patients with Colorectal Cancer (CRC), metastatic CRC cure rates have not improved due to high rates of oxaliplatin resistance and toxicity. There is an urgent need for effective tools to prevent and treat CRC and reduce morbidity and mortality of CRC patients. Exploring the effects of bioactive peptides on the antitumor to CRC was of vital importance to the clinical application. Objective: This study aimed to investigate the therapeutic impact of Anticancer Bioactive Peptides (ACBP) on anticancer effect of oxaliplatin (LOHP) in human colorectal cancer xenografts models in nude mice. Methods: HCT-116 cells were cultured in vitro via CCK-8 assays and the absorbance was measured at 450 nm. Apoptosis and cell cycle were assessed by Flow Cytometry (FCM) in vitro. HCT-116 human colorectal cancer cells inoculated subcutaneously in nude mice of treatment with PBS (GG), ACBP, LOHP, ACBP+LOHP (A+L) in vivo. The quality of life was assessed by dietary amount of nude mice, the weight of nude mice, inhibition rates, tumor weight and tumor volume. Immunohistochemistry and RT-qPCR method was conducted to determine the levels of apoptosisregulating proteins/genes in transplanted tumors. Results: ACBP induced substantial reductions in viable cell numbers and apoptosis of HCT116 cells in combined with LOHP in vitro. Compared with the control GG group, ACBP combined low dose oxaliplatin (U) group demonstrated significantly different tumor volume, the rate of apoptosis, the expression levels of Cyt-C, caspase-3,8,9 proteins and corresponding RNAs (P<0.05). The expression of pro-apoptotic proteins in the cytoplasm around the nucleus was significantly enhanced by ACBP. Short term intermittent use of ACBP alone indicted a certain inhibitory effect on tumor growth, and improve the quality of life of tumor bearing nude mice. Conclusion: ACBP significantly increased the anti-cancer responses of low dose oxaliplatin (L-LOHP), thus, significantly improving the quality of life of tumor-bearing nude mice.

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A Call for a European Prohibition of Monoclonal Antibody Production by the Ascites Procedure in Laboratory Animals

Alternatives to Laboratory Animals ◽

10.1177/026119299802600414 ◽

1998 ◽

Vol 26 (4) ◽

pp. 523-536

Author(s):

Coenraad F.M. Hendriksen

Keyword(s):

Monoclonal Antibody ◽

Laboratory Animals ◽

Induction Method ◽

Therapeutic Application ◽

Monoclonal Antibody Production ◽

In Vitro Methods ◽

Appeals Process ◽

Core Facilities ◽

Mab Production

Monoclonal antibodies (mAbs) are particularly valuable in therapeutics and research. Unfortunately, one of the most familiar methods of producing mAbs, the ascites induction method, causes pain and distress to the animals used. In most cases, non-animal or in vitro alternatives can be employed to reduce or eliminate the use of animals for mAb production. Prohibition of the use of animals in the production of mAbs is recommended, except when the replacement in vitro methods prove to be insufficient, and in a limited number of other well-documented cases, such as an exceptional need for an emergency therapeutic application. A total ban on the use of animals for mAb production is impractical and it is imperative that an appeals process should accompany the prohibition. The need for the establishment of core facilities for in vitro mAb production is emphasised.

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Cryopreservation of Whole Tumor Biopsies from Rectal Cancer Patients Enable Phenotypic and In Vitro Functional Evaluation of Tumor-Infiltrating T Cells

Cancers ◽

10.3390/cancers13102428 ◽

2021 ◽

Vol 13 (10) ◽

pp. 2428

Author(s):

Frank Liang ◽

Azar Rezapour ◽

Peter Falk ◽

Eva Angenete ◽

Ulf Yrlid

Keyword(s):

Rectal Cancer ◽

T Cells ◽

Γδ T Cells ◽

T Cell Subsets ◽

Functional Evaluation ◽

Mait Cells ◽

Ifn Γ ◽

Tumor Biopsies

TILs comprise functionally distinct conventional and unconventional T cell subsets and their role in responses to CRC treatments is poorly understood. We explored recovery of viable TILs from cryopreserved tumor biopsies of (chemo)-radiated patients with rectal cancer to establish a platform for retrospective TIL analyses of frozen tumors from pre-selected study cohorts. Frequencies of TIL subsets and their capacity to mount IFN-γ responses in cell suspensions of fresh vs. cryopreserved portions of the same tumor biopsies were determined for platform validation. The percentages and proportions of CD4+ TILs and CD8+ cytotoxic T lymphocytes (CTLs) among total TILs were not affected by cryopreservation. While recovery of unconventional γδ T cells and mucosal-associated invariant T cells (MAIT cells) was stable after cryopreservation, the regulatory T cells (Tregs) were reduced, but in sufficient yields for quantification. IFN-γ production by in vitro-stimulated CD4+ TILs, CTLs, γδ T cells, and MAIT cells were proportionally similar in fresh and cryopreserved tumor portions, albeit the latter displayed lower levels. Thus, the proposed platform intended for TIL analyses on cryopreserved tumor biobank biopsies holds promises for studies linking the quantity and quality of TIL subsets with specific clinical outcome after CRC treatment.

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