-
Experimental Animals 2012The common marmoset (Callithrix jacchus) is a species of New World monkeys. Because of its ease of maintenance and breeding in laboratories, use of the marmoset is... (Review)
Review
The common marmoset (Callithrix jacchus) is a species of New World monkeys. Because of its ease of maintenance and breeding in laboratories, use of the marmoset is growing rapidly in biomedical research. In neuroscience, the marmosets are attracting more attention, since they have the developed cerebral cortex which plays a key role in higher brain functions. In this review on neuroscience research using the marmoset, experimental techniques developed in our laboratory are summarized. We introduce artificial rearing of neonates, stereotaxic surgery, neuroanatomy including virtual microscopy based on the Internet technology, behavioral study using a large number of marmosets, and primary neuron culture study.
Topics: Animal Husbandry; Animals; Brain; Callithrix; Neurons; Neurosciences; Organ Culture Techniques; Stereotaxic Techniques
PubMed: 22850638
DOI: 10.1538/expanim.61.389 -
Integrative Biology : Quantitative... Jun 2016Human development and disease are challenging to study because of lack of experimental accessibility to in vivo systems and the complex nature of biological processes.... (Review)
Review
Human development and disease are challenging to study because of lack of experimental accessibility to in vivo systems and the complex nature of biological processes. For these reasons researchers turn to the use of model systems, ranging in complexity and scale from single cells to model organisms. While the use of model organisms is valuable for studying physiology and pathophysiology in an in vivo context and for aiding pre-clinical development of therapeutics, animal models are costly, difficult to interrogate, and not always equivalent to human biology. For these reasons, three-dimensional (3D) cell cultures have emerged as an attractive model system that contains key aspects of in vivo tissue and organ complexity while being more experimentally tractable than model organisms. In particular, organ-on-a-chip and organoid models represent orthogonal approaches that have been able to recapitulate characteristics of physiology and disease. Here, we review advances in these two categories of 3D cultures and applications in studying development and disease. Additionally, we discuss development of key technologies that facilitate the generation of 3D cultures, including microfluidics, biomaterials, genome editing, and imaging technologies.
Topics: Animals; Batch Cell Culture Techniques; Bioartificial Organs; Humans; Lab-On-A-Chip Devices; Organ Culture Techniques; Organoids; Printing, Three-Dimensional; Technology Assessment, Biomedical
PubMed: 27156572
DOI: 10.1039/c6ib00039h -
Neuron Oct 2018Disorders of the nervous system are challenging to study and treat due to the relative inaccessibility of functional human brain tissue for research. Stem cell-derived... (Review)
Review
Disorders of the nervous system are challenging to study and treat due to the relative inaccessibility of functional human brain tissue for research. Stem cell-derived 3D human brain organoids have the potential to recapitulate features of the human brain with greater complexity than 2D models and are increasingly being applied to model diseases affecting the central nervous system. Here, we review the use of human brain organoids to investigate neurological and psychiatric (neuropsychiatric) disorders and how this technology may ultimately advance our biological understanding of these conditions.
Topics: Brain Diseases; Humans; Organ Culture Techniques; Organoids
PubMed: 30359604
DOI: 10.1016/j.neuron.2018.10.007 -
Atherosclerosis Jan 2016Vascular calcification is a characteristic feature of aging, atherosclerosis, diabetes mellitus, and end-stage renal disease. The use of organ culture provides...
Vascular calcification is a characteristic feature of aging, atherosclerosis, diabetes mellitus, and end-stage renal disease. The use of organ culture provides complementary information that may bridge the gap between traditional cell culture and animal models, and establishes easily controlled experimental conditions. Therefore, we investigated whether organ culture of the aorta could be used as a model of vascular calcification, applying it to animal models of other conditions. Thoracic aortas were dissected from C57BL/6 mice and cultured. To induce vascular calcification, stimulation with inorganic phosphate (Pi) was performed. Morphometric assessment of medial calcium deposition was quantitatively performed, and the amount of dissolved calcium was measured. Pi-stimulation induced calcium deposition in medial layers in a time- and dose-dependent manner. To investigate the phenotypic change of vascular smooth muscle cells (VSMC), the expression of Runx2, osterix, osteocalcin, and ALP activity were determined. Finally, to investigate the influence of Pi-stimulation on the cultured aorta in other models, aortas from streptozotocin (STZ)-induced diabetic mice, aged mice, and Sirt1 knockout (+/-) mice were dissected. These cultures showed a greater tendency for aortic calcification by Pi-stimulation than did control cultures. These results indicate that organ culture of the aorta from mice reflects the state of calcification and suggests that this model will be useful to explore the molecular mechanisms of vascular calcification and the pathology of senescence.
Topics: Animals; Aorta, Thoracic; Apoptosis; Calcium; Cells, Cultured; Disease Models, Animal; Immunoblotting; Male; Mice; Mice, Inbred C57BL; Muscle, Smooth, Vascular; Organ Culture Techniques; Vascular Calcification
PubMed: 26584139
DOI: 10.1016/j.atherosclerosis.2015.11.005 -
Poultry Science Apr 2019The objective of this research was to examine the effects of prolactin (PRL) on the lipid synthesis of organ-cultured pigeon crops in vitro. In experiment 1, the...
The objective of this research was to examine the effects of prolactin (PRL) on the lipid synthesis of organ-cultured pigeon crops in vitro. In experiment 1, the histology, activities of enzymes, and expression of genes involved in metabolism and apoptosis of organ-cultured pigeon crops were analyzed over a 7-d culture period. The results showed that cultured crops maintained their structural integrity for up to 3 d in vitro. Beyond 3 d, caspase-3 activity and Bak1 gene expression increased with day of culture, whereas the activities of succinate dehydrogenase, Na+-K+-ATPase, Ca2+-Mg2+-ATPase, total ATPase, and gene expression of Bcl-2 and CK-19 diminished (P < 0.05). In experiment 2, the crops were cultured for 24, 36, and 48 h in medium containing 0, 25, or 50 ng/mL PRL, respectively, and the accumulation of lipid droplets, lipid content, and expression of fatty acid transportation- and lipogenesis-related genes were analyzed. The results showed that the crops with PRL supplements showed higher amounts of lipid droplets than those of the controls, and the droplets were mainly located in the basal nutritive layer in response to PRL. The efficacy of inducing lipid accumulation increased as the concentration of PRL increased. Crops with 50 ng/mL PRL incubated for 36 h displayed the maximal lipid content. Increasing the concentration of PRL from 0 to 50 ng/mL resulted in a dose-dependent increase in the expression of acetyl-CoA carboxylase, fatty acid synthase, fatty acid translocase, fatty acid binding protein 5, acyl-CoA binding protein, and peroxisome proliferator-activated receptor γ genes after incubation for 36 h (P < 0.05). Therefore, our results indicated that the organ-cultured pigeon crops maintained good viability for up to 3 d in vitro. Furthermore, PRL induced the lipid synthesis of organ-cultured pigeon crops in a dose- and time-dependent manner, which was related to the increased expression of genes involved in fatty acid transportation and lipogenesis.
Topics: Animals; Avian Proteins; Columbidae; Crop, Avian; Lipogenesis; Organ Culture Techniques; Prolactin
PubMed: 30590797
DOI: 10.3382/ps/pey540 -
Trends in Immunology Aug 2017Studying the interactions between commensal microbes and host intestinal tissue networks is challenging due to the complexity and inaccessibility of the system. A recent...
Studying the interactions between commensal microbes and host intestinal tissue networks is challenging due to the complexity and inaccessibility of the system. A recent study reports a novel organ culture system that will enhance our ability to dissect these interactions.
Topics: Bioengineering; Intestines; Mucous Membrane; Nervous System; Organ Culture Techniques
PubMed: 28684208
DOI: 10.1016/j.it.2017.06.007 -
Developmental Biology Dec 2016The ability to model human brain development in vitro represents an important step in our study of developmental processes and neurological disorders. Protocols that... (Review)
Review
The ability to model human brain development in vitro represents an important step in our study of developmental processes and neurological disorders. Protocols that utilize human embryonic and induced pluripotent stem cells can now generate organoids which faithfully recapitulate, on a cell-biological and gene expression level, the early period of human embryonic and fetal brain development. In combination with novel gene editing tools, such as CRISPR, these methods represent an unprecedented model system in the field of mammalian neural development. In this review, we focus on the similarities of current organoid methods to in vivo brain development, discuss their limitations and potential improvements, and explore the future venues of brain organoid research.
Topics: Brain; Humans; Models, Neurological; Neurodevelopmental Disorders; Organ Culture Techniques; Organogenesis; Organoids
PubMed: 27402594
DOI: 10.1016/j.ydbio.2016.06.037 -
International Journal of Molecular... Nov 2022Corneal transplantation is one of the most common forms of tissue transplantation worldwide. Donor corneal tissue used in transplantation is provided by eye banks, which...
Corneal transplantation is one of the most common forms of tissue transplantation worldwide. Donor corneal tissue used in transplantation is provided by eye banks, which store the tissue in culture medium after procurement. To date, the effects of cell culture on human corneal tissue have not been fully elucidated. Using the 3' RNA sequencing method for massive analysis of cDNA ends (MACE), we show that cultivation of corneal tissue leads to significant changes in a variety of molecular processes in human corneal tissue that go well beyond aspects of previously known culture effects. Functionally grouped network analysis revealed nine major groups of biological processes that were affected by corneal organ culture, among them keratinization, hypoxia, and angiogenesis, with genes from each group being affected by culture time. A cell type deconvolution analysis revealed significant modulations of the corneal immune cell profile in a time dependent manner. The results suggest that current culture conditions should be further refined and that prolonged cultivation may be detrimental. Recently, we showed that MACE enables transcriptional profiling of formalin-fixed and paraffin-embedded (FFPE) conjunctival tissue with high accuracy even after more than 10 years of storage. Here we demonstrate that MACE provides comparable results for native and FFPE corneal tissue, confirming that the technology is suitable for transcriptome analysis of a wide range of archived diseased corneal samples stored in histological archives. Finally, our data underscore the feasibility of bioinformatics cell-type enrichment analysis in bulk RNA-seq data to profile immune cell composition in fixed and archived corneal tissue samples, for which RNA-seq analysis of individual cells is often not possible.
Topics: Humans; Organ Culture Techniques; Organ Preservation; Eye Banks; Tissue Donors; Cornea; DNA, Complementary
PubMed: 36498835
DOI: 10.3390/ijms232314507 -
European Cells & Materials Jul 2014Annulus fibrosus (AF) defects from annular tears, herniation, and discectomy procedures are associated with painful conditions and accelerated intervertebral disc (IVD)...
Annulus fibrosus (AF) defects from annular tears, herniation, and discectomy procedures are associated with painful conditions and accelerated intervertebral disc (IVD) degeneration. Currently, no effective treatments exist to repair AF damage, restore IVD biomechanics and promote tissue regeneration. An injectable fibrin-genipin adhesive hydrogel (Fib-Gen) was evaluated for its performance repairing large AF defects in a bovine caudal IVD model using ex vivo organ culture and biomechanical testing of motion segments, and for its in vivo longevity and biocompatibility in a rat model by subcutaneous implantation. Fib-Gen sealed AF defects, prevented IVD height loss, and remained well-integrated with native AF tissue following approximately 14,000 cycles of compression in 6-day organ culture experiments. Fib-Gen repair also retained high viability of native AF cells near the repair site, reduced nitric oxide released to the media, and showed evidence of AF cell migration into the gel. Biomechanically, Fib-Gen fully restored compressive stiffness to intact levels validating organ culture findings. However, only partial restoration of tensile and torsional stiffness was obtained, suggesting opportunities to enhance this formulation. Subcutaneous implantation results, when compared with the literature, suggested Fib-Gen exhibited similar biocompatibility behaviour to fibrin alone but degraded much more slowly. We conclude that injectable Fib-Gen successfully sealed large AF defects, promoted functional restoration with improved motion segment biomechanics, and served as a biocompatible adhesive biomaterial that had greatly enhanced in vivo longevity compared to fibrin. Fib-Gen offers promise for AF repairs that may prevent painful conditions and accelerated degeneration of the IVD, and warrants further material development and evaluation.
Topics: Animals; Bioreactors; Cattle; Chondrogenesis; Compressive Strength; Fibrin Tissue Adhesive; Hydrogels; Intervertebral Disc; Intervertebral Disc Degeneration; Iridoids; Nitric Oxide; Organ Culture Techniques; Rats; Rats, Sprague-Dawley; Regeneration; Stress, Mechanical; Tensile Strength; Torque
PubMed: 25036053
DOI: 10.22203/ecm.v028a03 -
Journal of Ocular Pharmacology and... Nov 2020Corneal endothelial dystrophies are characterized by endothelial cell loss and dysfunction. Recent evidence suggests that corneal endothelial cells (CECs) can...
Corneal endothelial dystrophies are characterized by endothelial cell loss and dysfunction. Recent evidence suggests that corneal endothelial cells (CECs) can regenerate although they do not do so under normal conditions. This work sought to test whether CECs can be stimulated to proliferate in organ culture by wounding and/or by treatment with the engineered human fibroblast growth factor 1 (FGF1) derivative TTHX1114. Human donor corneas obtained from eye banks were maintained in organ culture in the presence or absence of TTHX1114. Wounds in the corneas were created by quartering the corneas. The CEC monolayer was identified as a regular layer by Hoechst staining of the nuclear DNA with cell outlines delineated by immunohistochemical identification of ZO-1. Nuclei and nuclei incorporating 5-ethynyl-2'-deoxyuridine (EdU) were counted using ImageJ. CECs in normal corneas in undisturbed monolayers had low, but measurable, rates of proliferation. CECs at the edge of a wound had higher rates of proliferation, probably due to the release of contact inhibition. TTHX1114 increased proliferation at wound edges. After 7 days of culture, proliferating CECs formed contiguous groups of labeled cells that did not migrate away from one another. TTHX1114-treated cells, including the EdU labeled proliferating cells, retained normal morphology, including cell/cell junction ZO-1 staining. Proliferation of CECs in organ-cultured corneas is low, but can be stimulated by wounding or by the administration of TTHX1114 with the effects of each being additive. The CEC monolayer appears to have a population of progenitor cells that are susceptible to stimulation.
Topics: Adult; Aged; Cell Proliferation; Endothelium, Corneal; Female; Fibroblast Growth Factor 1; Humans; Male; Middle Aged; Organ Culture Techniques; Protein Engineering; Wound Healing
PubMed: 32735473
DOI: 10.1089/jop.2019.0119