Use of stem cells as alternative methods to animal experimentation in predictive toxicology

2019 ◽  
Vol 105 ◽  
pp. 15-29 ◽  
Author(s):  
Tae-Won Kim ◽  
Jeong-Hwan Che ◽  
Jun-Won Yun
Keyword(s):  
Stem Cells ◽  
Animal Experimentation ◽  
Alternative Methods ◽  
Predictive Toxicology

scholarly journals Timing of Peripheral Blood Stem Cell Yield: Comparison of Alternative Methods with the Classic Method for CD34+Cell Determination

2014 ◽  
Vol 2014 ◽  
pp. 1-13 ◽  
Author(s):  
I. Fatorova ◽  
M. Blaha ◽  
M. Lanska ◽  
D. Vokurkova ◽  
V. Rezacova ◽  
...  
Keyword(s):  
Stem Cells ◽  
Peripheral Blood ◽  
Reference Method ◽  
Roc Curves ◽  
Alternative Methods ◽  
Optimal Timing ◽  
Hematopoietic Tissue ◽  
Peripheral Blood Stem Cells ◽  
Hematopoietic Stem ◽  
Blood Stem Cells

Hematopoietic stem cells (HSCs), still represent a certain mystery in biology, have a unique property of dividing into equal cells and repopulating the hematopoietic tissue. This potential enables their use in transplantation treatments. The quality of the HSC grafts for transplantation is evaluated by flow cytometric determination of the CD34+cells, which enables optimal timing of the first apheresis and the acquisition of maximal yield of the peripheral blood stem cells (PBSCs). To identify a more efficient method for evaluating CD34+cells, we compared the following alternative methods with the reference method: hematopoietic progenitor cells (HPC) enumeration (using the Sysmex XE-2100 analyser), detection of CD133+cells, and quantification of aldehyde dehydrogenase activity in the PBSCs. 266 aphereses (84 patients) were evaluated. In the preapheretic blood, the new methods produced data that were in agreement with the reference method. The ROC curves have shown that for the first-day apheresis target, the optimal predictive cut-off value was 0.032 cells/mL for the HPC method (sensitivity 73.4%, specificity 69.3%). HPC method exhibited a definite practical superiority as compared to other methods tested. HPC enumeration could serve as a supplementary method for the optimal timing of the first apheresis; it is simple, rapid, and cheap.

Investigation of the Stem Cells Used in Modeling Human Placental Trophoblast

2021 ◽  
Author(s):  
Lulu Ji ◽  
Lin Wang
Keyword(s):  
Stem Cells ◽  
Molecular Mechanisms ◽  
Embryonic Stem ◽  
Cell Lineage ◽  
Cell Types ◽  
Model Systems ◽  
Alternative Methods ◽  
Laboratory Animals ◽  
Placental Development ◽  
Induced Pluripotent

Human placenta is vital for fetal development, and act as an interface between the fetus and the expecting mother. Abnormal placentati on underpins various pregnancy complications such as miscarriage, pre-eclampsia and intrauterine growth restriction. Despite the important role of placenta, the molecular mechanisms governing placental formation and trophoblast cell lineage specification is poorly understand. It is mostly due to the lack of appropriate model system. The great various in placental types across mammals make it limit for the use of laboratory animals in studying human placental development. However, over the past few years, alternative methods have been employed, including human embryonic stem cells, induced pluripotent stem cells, human trophoblast stem cell, and 3-dimensional organoids. Herein, we summarize the present knowledge about human development, differentiated cell types in the trophoblast epithelium and current human placental trophoblast model systems.

scholarly journals Physiological aspects of cardiac tissue engineering

2012 ◽  
Vol 303 (2) ◽  
pp. H133-H143 ◽  
Author(s):  
Thomas Eschenhagen ◽  
Alexandra Eder ◽  
Ingra Vollert ◽  
Arne Hansen
Keyword(s):  
Tissue Engineering ◽  
Stem Cells ◽  
Cardiac Myocyte ◽  
Cardiac Tissue ◽  
Cell Sheet ◽  
Three Dimensional ◽  
Embryonic Stem ◽  
Basic Research ◽  
Cardiac Tissue Engineering ◽  
Predictive Toxicology

Cardiac tissue engineering aims at repairing the diseased heart and developing cardiac tissues for basic research and predictive toxicology applications. Since the first description of engineered heart tissue 15 years ago, major development steps were directed toward these three goals. Technical innovations led to improved three-dimensional cardiac tissue structure and near physiological contractile force development. Automation and standardization allow medium throughput screening. Larger constructs composed of many small engineered heart tissues or stacked cell sheet tissues were tested for cardiac repair and were associated with functional improvements in rats. Whether these approaches can be simply transferred to larger animals or the human patients remains to be tested. The availability of an unrestricted human cardiac myocyte cell source from human embryonic stem cells or human-induced pluripotent stem cells is a major breakthrough. This review summarizes current tissue engineering techniques with their strengths and limitations and possible future applications.

Red Marrow Dosimetry and Stem Cell Reinfusion in High Dose 90Y - Ibritumomab Tiuxetan.

Blood ◽  
2008 ◽  
Vol 112 (11) ◽  
pp. 2187-2187 ◽  
Author(s):  
Anna Vanazzi ◽  
Daniele Laszlo ◽  
Marta Cremonesi ◽  
Chiara Maria Grana ◽  
Stefano Papi ◽  
...  
Keyword(s):  
Stem Cells ◽  
Bone Marrow ◽  
Image Analysis ◽  
Activity Concentration ◽  
Activity Level ◽  
Alternative Methods ◽  
Ibritumomab Tiuxetan ◽  
Time Activity ◽  
Bone Marrow Biopsies ◽  
Red Marrow

Abstract BACKGROUND: Red marrow (RM) is the critical organ in Radioimmunotherapy (RIT) with 90Y-ibritumomab tiuxetan, especially at myeloablative activities. Different methods for the evaluation of the adsorbed dose in RM have been investigated. The blood method relates the blood time-activity curve to RM by a factor that accounts for the activity concentration ratio between RM and blood. Alternative dosimetric approaches including image analysis and marrow aspirations (MA) were introduced for RM dose comparison in order to collect information about the dose delivered and to verify the correct time of Autologous Stem Cells Transplantation (ASCT). PATIENTS AND METHODS: Twenty-six patients affected by resistant/refractory B-cell Non Hodgkin Lymphoma (NHL) were enrolled in a phase I/II study to determine the feasibility and safety of High Activity (HA) of 90Y-ibritumomab tiuxetan followed by ASC support. Three 90Y-ibritumomab tiuxetan activity levels were planned: 30 MBq/kg (4 patients), 45 MBq/kg (4 patients), 56 MBq/kg (18 patients). One week before treatment all patients underwent dosimetry with 111Inibritumomab tiuxetan. The activity concentration was measured in 9 serial blood samples up to 7 days post injection (pi) and planar images in all patients, in MA in 8 out of 18 patients planned at 56 MBq/kg (4 patients at 160 h pi; 4 patients on day planned for ASCT, i.e. day 0). ASC were reinfused 13 days after 90Y-ibritumomab tiuxetan. ASCT was considered at low risk when the adsorbed dose to reinfused stem cells (rSC) (i.e. dose to RM at time of ASCT) was < 50 mGy. Engraftment was considered delayed if on day 28 from reinfusion ANC did not reach 1.0x109/L or PLT were not ≥ 20.0x109/L. RESULTS: A median of 4,25x106/kg (1,45–20x106/kg) CD34+ cells were reinfused for each patient. A delay of engraftment occurred in only 1 patient who received 56 MBq/kg with a baseline ANC count of 0.20x109/L. Bone marrow biopsies were negative for disease localization in all patients at baseline, but 10 out of 26 had shown prior lymphoma infiltration. Considering the blood method the adsorbed dose to rSC resulted < 50 mGy in all patients with a median adsorbed dose of 11 mGy (4 – 28 mGy). ASC would be reinfused earlier in all patients except one. In particular, the 1st day with a dose to rSC < 50 mGy ranged from day -9 to day -5 at 30 MBq/kg; from day -8 to day -4 at 45 MBq/kg; from day -9 to day 0 at 56 MBq/kg, with a median value on day -3. We observed a possible correlation between total activity administered, dose delivered to RM and dose detectable on day of reinfusion, with higher doses to RM and rSC for those patients planned to received the highest activity. We also evaluated the dose to rSC if reinfusion would be performed 7 days after RIT: only 6 out of 26 patients showed a dose to SC < 50 mGy (2 at the 1st activity level, 3 at the 2nd and 1 at the 3rd activity level). Considering the MA method, the median adsorbed dose to the rSC resulted 25 mGy (9 - 69 mGy): in 2 out of 8 patients evaluated the adsorbed dose to the rSC was > 50 mGy on day 0 (69 and 51 mGy respectively). Even if ASC would be reinfused earlier in 5 out of 8 patients, the first day with a dose to rSC < 50 mGy ranged from day -3 to day +1. Despite bone marrow biopsy being negative in all patients at baseline, scintigraphic images (L2–L4 uptake) showed spine marrow uptake in almost all patients. CONCLUSIONS: Although alternative methods investigated - MA and imaging - are affected by uncertain factors (blood contamination of MA, difficult L2-L4 uptake image quantification), both indicate comparable results, about 2.5-folds higher, suggesting that the blood method provides an underestimate of RM dose. ASCT performed 13 days after HA 90Y-ibritumomab tiuxetan is generally safe. We suggest caution in anticipating the timing of ASCT with respect to 90Y-ibritumomab tiuxetan in those patients planned to receive an activity > 45 MBq/kg: alternative dosimetric approaches including image analysis and MA should be considered.

scholarly journals Human Organoids for Predictive Toxicology Research and Drug Development

2021 ◽  
Vol 12 ◽  
Author(s):  
Toshikatsu Matsui ◽  
Tadahiro Shinozawa
Keyword(s):  
Stem Cells ◽  
Drug Development ◽  
Disease Modeling ◽  
Cell Types ◽  
Underlying Disease ◽  
Future Research ◽  
Specific Cell ◽  
Predictive Toxicology

Organoids are three-dimensional structures fabricated in vitro from pluripotent stem cells or adult tissue stem cells via a process of self-organization that results in the formation of organ-specific cell types. Human organoids are expected to mimic complex microenvironments and many of the in vivo physiological functions of relevant tissues, thus filling the translational gap between animals and humans and increasing our understanding of the mechanisms underlying disease and developmental processes. In the last decade, organoid research has attracted increasing attention in areas such as disease modeling, drug development, regenerative medicine, toxicology research, and personalized medicine. In particular, in the field of toxicology, where there are various traditional models, human organoids are expected to blaze a new path in future research by overcoming the current limitations, such as those related to differences in drug responses among species. Here, we discuss the potential usefulness, limitations, and future prospects of human liver, heart, kidney, gut, and brain organoids from the viewpoints of predictive toxicology research and drug development, providing cutting edge information on their fabrication methods and functional characteristics.

scholarly journals COMPARISON OF THERAPEUTIC EFFECTS OF DIFFERENT METHODS OF ADMINISTRATION OF MEZENCHIMAL STEM CELLS TO MICE WITH PREMATURE OVARIAN INSUFFICIENCY

2021 ◽  
Vol 76 (2) ◽  
pp. 35-40
Author(s):  
M. I. Kozub ◽  
K. P. Skybina ◽  
I. B. Musatova ◽  
O. V. Prokopiuk ◽  
S. M. Gramatiuk ◽  
...  
Keyword(s):  
Stem Cells ◽  
Sexual Function ◽  
Ovarian Function ◽  
Hormone Replacement ◽  
Morphological Structure ◽  
Premature Ovarian Insufficiency ◽  
Alternative Methods ◽  
Therapeutic Effects ◽  
Ovarian Insufficiency ◽  
Anticancer Chemotherapy

Premature ovarian insufficiency (POI) is condition of female hypogonadism. One of the reasons for the development of POI is anticancer chemotherapy. Currently, hormone replacement therapy is considered the most effective method of treatment POI, which, however, had a number of side effects, requires constant administration of pharmaceuticals, did not restore fertility and correct pathological changes in target organs. Therefore, the search for alternative methods of treatment based on the use of stem cells is promising. The aim of work was to compare the therapeutic effects of different methods of administration of mesenchymal stem cells isolated from adipose tissue (AT MSCs) in restoring sexual function, as well as the behavior of mice with chemotherapy-induced premature ovarian failure. Materials and methods. The investigation was carried out on BALB/c mice with a POI model induced by Cyclophosphamide and Busulfan. A histological examination of preparations of the ovaries and uterus was carried out. We studied the changes in the weight of mice, ovarian function, sexual activity, as well as the behavior of animals in anxiolytic and zoosocial tests after treatment with AT MSCs with intraperitoneal and intraovarian routes of administration. Results and discussion. It was shown that the effectiveness of the use of AT MSCs depends on the method of their administration at the treatment of POI induced by chemotherapy. More rapid and complete restoration of the morphological structure, sexual function, and behavioral responses were observed in mice with POI treated by intraovarial administration of AT MSCs.  

388 OPTIMIZATION OF INDUCED PLURIPOTENT STEM CELL GENERATION FROM BOVINE FIBROBLASTS

2010 ◽  
Vol 22 (1) ◽  
pp. 350
Author(s):  
M. L. Lim ◽  
H. Sumer ◽  
J. Liu ◽  
P. J. Verma
Keyword(s):  
Stem Cells ◽  
Cell Density ◽  
Pluripotent Stem Cells ◽  
Ips Cells ◽  
Alternative Methods ◽  
Packaging Cell Line ◽  
A Cell ◽  
Reprogramming Factors ◽  
Induced Pluripotent ◽  
Adult Fibroblasts

Difficulties associated with isolation and culturing bovine embryonic stem (ES) cells has led to the exploration of alternative methods for generating pluripotent stem cells. The viral delivery of reprogramming factors Oct4, Sox2, cMyc, and Klf4 has resulted in generation of induced pluripotent stem cells (iPS) in rodent, human, rhesus monkey, and pig somatic cells. In the current study, we improved the efficiency of retroviral transduction of bovine adult fibroblasts (BAF) for the generation of bovine iPS cells. Bovine adult fibroblasts were transduced with 4 human factors: Oct 4, Sox 2, cMyc, and Klf4. To determine transfection efficiency, pMXs-GFP was used as a reporter. The effect of change in cell density of the Platinum A retroviral packaging cell line (Plat A), cell densities of the target BAF, and infection regimes on the transfection rates was examined. A reduction in Plat A cell density from 8 × 106 to 2 × 106 did not alter transfection rates. Reduced target cell density from 4 × 105 to 4 × 104 (10-fold) improved the transfection rates from 0.31 to 7.06%, P < 0.001 (n = 3). Subjecting the BAF to 2 sequential rounds of viral transduction further improved the transfection rates to 13.88%, P < 0.001 (n = 3). These preliminary results suggest that optimizing the density of target cells can greatly improve transduction outcomes. Following viral induction with the 4 reprogramming factors, putative bovine iPS colonies were observed when the transfection rate was >1%. The putative bovine iPS cells were cultured in alpha-minimal essential medium supplemented with 20% FCS and 10 ng mL-1 human leukemia inhibitory factor. These putative bovine iPS colonies had mouse ES-like morphology, were multilayered, and had high nucleus-to-cytoplasmic ratio. They stained positive for alkaline phosphatase activity. The colonies were manually passaged onto mitomycin C-inactivated mouse embryonic fibroblasts every 5 to 7 days but could only be expanded for a limited number of passages. Other strategies are currently being explored to improve stable reprogramming of BAF such as epigenetic modification of cells, lentivirus-mediated transduction, and investigation of media suitable to maintain putative bovine iPS colonies for further characterization including RT-PCR or immunohistochemical detection for pluripotent markers and in vivo differentiation ability. Acknowledgments are given to Dairy Australia.

Saviour-Sibling and the Psychological, Ethical and Judicial Issues that It Creates: Should English and French Legislators Close the Pandora’s Box?

2011 ◽  
Vol 18 (3) ◽  
pp. 293-303 ◽  
Author(s):  
Allane Madanamoothoo
Keyword(s):  
Stem Cells ◽  
Bone Marrow ◽  
Umbilical Cord ◽  
Genetic Disease ◽  
Ethical Issues ◽  
Test Tube ◽  
Alternative Methods ◽  
Official Journal ◽  
Human Fertilisation ◽  
Biomedical Field

AbstractSince the advent of test-tube babies, advances in the biomedical field have risen steadily. In parallel, the scientific body has never since ceased to debate the ethical issues that they arise. This is particularly the case regarding saviour-sibling. Saviour-sibling refers to a child who is conceived to cure an older brother or sister suffering from a serious family genetic disease. Therefore, it is meant to give birth to a child who will provide stem-cells taken from the umbilical cord or bone marrow afterwards, to treat an elder sick sibling. In England, this practice has been explicitly allowed by the new Human Fertilisation and Embryology Act 2008 under some strict conditions. In France, this practice, authorized by the Bioethics Law of August 2004 and confirmed by its decree of implementation published in the Official Journal on 23 December 2006, is also strictly regulated. This technique opens up new perspectives and enormous hope. Its legalisation is certainly justified by the suffering of the parents and to avoid that they travel to other States where it is permitted. However, it raises serious psychological ethical and judicial issues. Following an analysis of the English and French laws on saviour siblings, its controversial side will be highlighted, before concluding whether or not this new Pandora’s box which is saviour-sibling, should be closed and other alternative methods encouraged.

Sign in / Sign up

Export Citation Format

Share Document