scholarly journals In Vitro Comparison of 2D-Cell Culture and 3D-Cell Sheets of Scleraxis-Programmed Bone Marrow Derived Mesenchymal Stem Cells to Primary Tendon Stem/Progenitor Cells for Tendon Repair

2018 ◽  
Vol 19 (8) ◽  
pp. 2272 ◽  
Author(s):  
Chi-Fen Hsieh ◽  
Zexing Yan ◽  
Ricarda Schumann ◽  
Stefan Milz ◽  
Christian Pfeifer ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Cell Sheet ◽  
Tendon Repair ◽  
Three Dimensional ◽  
Cell Types ◽  
Tendon Tissue ◽  
Cell Sheets ◽  
Tendon Tissue Engineering ◽  
Dimensional Cell

The poor and slow healing capacity of tendons requires novel strategies to speed up the tendon repair process. Hence, new and promising developments in tendon tissue engineering have become increasingly relevant. Previously, we have established a tendon progenitor cell line via ectopic expression of the tendon-related basic helix-loop-helix (bHLH) transcription factor Scleraxis (Scx) in human bone marrow mesenchymal stem cells (hMSC-Scx). The aim of this study was to directly compare the characteristics of hMSC-Scx cells to that of primary human tendon stem/progenitors cells (hTSPCs) via assessment of self-renewal and multipotency, gene marker expression profiling, in vitro wound healing assay and three-dimensional cell sheet formation. As expected, hTSPCs were more naive than hMSC-Scx cells because of higher clonogenicity, trilineage differentiation potential, and expression of stem cell markers, as well as higher mRNA levels of several gene factors associated with early tendon development. Interestingly, with regards to wound healing, both cell types demonstrate a comparable speed of scratch closure, as well as migratory velocity and distance in various migration experiments. In the three-dimensional cell sheet model, hMSC-Scx cells and hTSPCs form compact tendinous sheets as histological staining, and transmission electron microscopy shows spindle-shaped cells and collagen type I fibrils with similar average diameter size and distribution. Taken together, hTSPCs exceed hMSC-Scx cells in several characteristics, namely clonogenicity, multipotentiality, gene expression profile and rates of tendon-like sheet formation, whilst in three-dimensional cell sheets, both cell types have comparable in vitro healing potential and collagenous composition of their three-dimensional cell sheets, making both cell types a suitable cell source for tendon tissue engineering and healing.

Models of Cell Interaction Based on Differential Adhesion

1984 ◽  
Vol 106 (1) ◽  
pp. 36-41 ◽  
Author(s):  
S. Childress
Keyword(s):  
Three Dimensional ◽  
Numerical Algorithms ◽  
Cell Types ◽  
Cell Interaction ◽  
Cell Aggregates ◽  
Biological Cells ◽  
Mechanical Interaction ◽  
Cell Sheets ◽  
Differential Adhesion ◽  
Dimensional Cell

Recent efforts to model the mechanical interaction of aggregates of biological cells are reviewed. Differential adhesion is discussed as a means of creating a field of stress equivalent to tension elements at interfaces between unlike cell types. Several numerical algorithms are described and applied to shortening and folding of cell sheets, three-dimensional monolayers, and two-dimensional cell aggregates.

Aligned Collagen Scaffolds Enhance Tenogenic Differentiation of Bone Marrow-Derived Stem Cells

2018 ◽  
Vol 773 ◽  
pp. 333-337 ◽  
Author(s):  
Ke Ke Zhang ◽  
Chang Liu ◽  
Yu Long Sun
Keyword(s):  
Stem Cells ◽  
Bone Marrow ◽  
Tendon Repair ◽  
Collagen Fibers ◽  
Collagen Scaffolds ◽  
Formidable Challenge ◽  
Tendon Tissue Engineering ◽  
Limited Function ◽  
Fiber Scaffolds

Tendon repair remains a formidable challenge in clinic, due to its limited function in long-term recovery. Decellularized native tendon extracellular matrix (ECM), which contains specific bioactive factors and a large number of collagen fibers, seeded with bone marrow-derived stem cells (BMSCs) has been found to promote tendon neogenesis. However, the effect of arrangements of natural collagen fibers on the differentiation of BMSCs remains unclear. Here, we prepared aligned collagen fiber scaffolds from rat tail tendon ECM to investigate the effect of collagen arrangements on the morphology, proliferation and differentiation of rat BMSCs in vitro. It was found that BMSCs were elongated and aligned along the collagen fibers on the aligned collagen fibers. Furthermore, the aligned collagen scaffolds significantly enhanced the expression of tenocyte markers TNMD and THBS4. The results indicated that aligned collagen scaffolds could induce the tenogenesis of BMSCs, which would provide an alternative approach for tendon tissue engineering.

scholarly journals Role of FBXW2 in explant cultures of bovine periosteum-derived cells

2021 ◽  
Vol 14 (1) ◽  
Author(s):  
Mari Akiyama
Keyword(s):  
Potential Role ◽  
Three Dimensional ◽  
Explant Culture ◽  
Cell Sheets ◽  
Explant Cultures ◽  
Cell Pellet ◽  
Cambium Layer ◽  
Dimensional Cell

Abstract Objective Bone regeneration is a potential technique for treating osteoporosis. A previous study reported that F-box and WD-40 domain-containing protein 2 (FBXW2) localized with osteocalcin in bovine periosteum after 5 weeks of explant culture. However, the osteoblastic functions of FBXW2 remain unclear. In this study, double-fluorescent immunostaining was used to investigate the potential role of FBXW2 and its relationship with osteocalcin. Results At day 0, FBXW2 was expressed in the cambium layer between the bone and periosteum, while osteocalcin was expressed in bone. After explant culture, changes in the periosteum were observed from weeks 1 to 7. At week 1, partial FBXW2 expression was seen with a small amount of osteocalcin. At week 2, a layer of FBXW2 was observed. From weeks 3 to 7, tube-like structures of FBXW and osteocalcin were observed, and periosteum-derived cells were released from the periosteum in areas where no FBXW2 was observed. Bovine periosteum-derived cells can form a three-dimensional cell pellet, because multilayered cell sheets are formed inside of the periosteum in vitro. It is shown that in results FBXW2 is produced in periosteal explants near sites where initial osteogenic activity is observed, suggesting that it may be involved in periosteal osteogenesis.

Multi-layered in vitro 3D-bone model via combination of osteogenic cell sheets with electrospun membrane interlayer

2021 ◽  
pp. 088532822110278
Author(s):  
Atakan Tevlek ◽  
Halil Murat Aydin
Keyword(s):  
Cell Sheet ◽  
Three Dimensional ◽  
Osteogenic Cell ◽  
Electrospun Membrane ◽  
Cell Sheets ◽  
Cell Sheet Engineering ◽  
Sulfated Glycosaminoglycan ◽  
Sgag Content ◽  
In Vitro Performance

In this study, it was aimed to present an approach for the development of multi-layered tissue engineering constructs by using cell sheet engineering. Briefly, MC3T3-E1 mouse pre-osteoblast cells were cultured in temperature-responsive plates (Nunc Upcell®) in the presence of osteogenic medium and the resulting cell sheets were laminated with electrospun poly(L-lactic acid) (PLLA) membranes to obtain viable three-dimensional, thick constructs. The constructs prepared without PLLA membranes were used as control. The cell viability and death in the resulting structures were investigated by microscopic and colorimetric methods. The in vitro performance of the structures was discussed comparatively. Alkaline phosphatase (ALP) activity, collagen and sulfated glycosaminoglycan (sGAG) content values were calculated. The presented approach shows potential for engineering applications of complex tissues with at least two or more microenvironments such as osteochondral, corneal or vascular tissues.

Correlated Studies of Cellular Interactions Using TEM, Freeze Etching, and SEM

1974 ◽  
Vol 32 ◽  
pp. 320-321
Author(s):  
J. P. Revel
Keyword(s):  
Developmental Stages ◽  
Three Dimensional ◽  
Cell Movement ◽  
Cellular Interactions ◽  
Serial Sections ◽  
Cell Sheets ◽  
Freeze Etching ◽  
Early Developmental

Movement of individual cells or of cell sheets and complex patterns of folding play a prominent role in the early developmental stages of the embryo. Our understanding of these processes is based on three- dimensional reconstructions laboriously prepared from serial sections, and from autoradiographic and other studies. Many concepts have also evolved from extrapolation of investigations of cell movement carried out in vitro. The scanning electron microscope now allows us to examine some of these events in situ. It is possible to prepare dissections of embryos and even of tissues of adult animals which reveal existing relationships between various structures more readily than used to be possible vithout an SEM.

Epithelial Organotypic Cultures: A Viable Model to Address Mechanisms of Carcinogenesis by Epitheliotropic Viruses

2018 ◽  
Vol 18 (4) ◽  
pp. 246-255 ◽  
Author(s):  
Lara Termini ◽  
Enrique Boccardo
Keyword(s):  
Three Dimensional ◽  
Cell Types ◽  
Experimental Models ◽  
Biological Research ◽  
Specific Gene ◽  
Specific Cell ◽  
Organotypic Cultures ◽  
Skin Physiology

In vitro culture of primary or established cell lines is one of the leading techniques in many areas of basic biological research. The use of pure or highly enriched cultures of specific cell types obtained from different tissues and genetics backgrounds has greatly contributed to our current understanding of normal and pathological cellular processes. Cells in culture are easily propagated generating an almost endless source of material for experimentation. Besides, they can be manipulated to achieve gene silencing, gene overexpression and genome editing turning possible the dissection of specific gene functions and signaling pathways. However, monolayer and suspension cultures of cells do not reproduce the cell type diversity, cell-cell contacts, cell-matrix interactions and differentiation pathways typical of the three-dimensional environment of tissues and organs from where they were originated. Therefore, different experimental animal models have been developed and applied to address these and other complex issues in vivo. However, these systems are costly and time consuming. Most importantly the use of animals in scientific research poses moral and ethical concerns facing a steadily increasing opposition from different sectors of the society. Therefore, there is an urgent need for the development of alternative in vitro experimental models that accurately reproduce the events observed in vivo to reduce the use of animals. Organotypic cultures combine the flexibility of traditional culture systems with the possibility of culturing different cell types in a 3D environment that reproduces both the structure and the physiology of the parental organ. Here we present a summarized description of the use of epithelial organotypic for the study of skin physiology, human papillomavirus biology and associated tumorigenesis.

scholarly journals Bone marrow stromal cells from MDS and AML patients show increased adipogenic potential with reduced Delta-like-1 expression

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Marie-Theresa Weickert ◽  
Judith S. Hecker ◽  
Michèle C. Buck ◽  
Christina Schreck ◽  
Jennifer Rivière ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Cell Fate ◽  
Stromal Cells ◽  
Adipogenic Differentiation ◽  
Cell Types ◽  
Bone Marrow Niche ◽  
Differentiation Potential ◽  
Hematopoietic Stem ◽  
Bm Niche

AbstractMyelodysplastic syndromes (MDS) and acute myeloid leukemia (AML) are clonal hematopoietic stem cell disorders with a poor prognosis, especially for elderly patients. Increasing evidence suggests that alterations in the non-hematopoietic microenvironment (bone marrow niche) can contribute to or initiate malignant transformation and promote disease progression. One of the key components of the bone marrow (BM) niche are BM stromal cells (BMSC) that give rise to osteoblasts and adipocytes. It has been shown that the balance between these two cell types plays an important role in the regulation of hematopoiesis. However, data on the number of BMSC and the regulation of their differentiation balance in the context of hematopoietic malignancies is scarce. We established a stringent flow cytometric protocol for the prospective isolation of a CD73+ CD105+ CD271+ BMSC subpopulation from uncultivated cryopreserved BM of MDS and AML patients as well as age-matched healthy donors. BMSC from MDS and AML patients showed a strongly reduced frequency of CFU-F (colony forming unit-fibroblast). Moreover, we found an altered phenotype and reduced replating efficiency upon passaging of BMSC from MDS and AML samples. Expression analysis of genes involved in adipo- and osteogenic differentiation as well as Wnt- and Notch-signalling pathways showed significantly reduced levels of DLK1, an early adipogenic cell fate inhibitor in MDS and AML BMSC. Matching this observation, functional analysis showed significantly increased in vitro adipogenic differentiation potential in BMSC from MDS and AML patients. Overall, our data show BMSC with a reduced CFU-F capacity, and an altered molecular and functional profile from MDS and AML patients in culture, indicating an increased adipogenic lineage potential that is likely to provide a disease-promoting microenvironment.

scholarly journals Human iPSC-Based Modeling of Central Nerve System Disorders for Drug Discovery

2021 ◽  
Vol 22 (3) ◽  
pp. 1203
Author(s):  
Lu Qian ◽  
Julia TCW
Keyword(s):  
Drug Discovery ◽  
Disease Modeling ◽  
Three Dimensional ◽  
Structural Complexity ◽  
Induced Pluripotent Stem Cell ◽  
Cell Types ◽  
Central Nerve System ◽  
Human Ipscs ◽  
Nerve System

A high-throughput drug screen identifies potentially promising therapeutics for clinical trials. However, limitations that persist in current disease modeling with limited physiological relevancy of human patients skew drug responses, hamper translation of clinical efficacy, and contribute to high clinical attritions. The emergence of induced pluripotent stem cell (iPSC) technology revolutionizes the paradigm of drug discovery. In particular, iPSC-based three-dimensional (3D) tissue engineering that appears as a promising vehicle of in vitro disease modeling provides more sophisticated tissue architectures and micro-environmental cues than a traditional two-dimensional (2D) culture. Here we discuss 3D based organoids/spheroids that construct the advanced modeling with evolved structural complexity, which propels drug discovery by exhibiting more human specific and diverse pathologies that are not perceived in 2D or animal models. We will then focus on various central nerve system (CNS) disease modeling using human iPSCs, leading to uncovering disease pathogenesis that guides the development of therapeutic strategies. Finally, we will address new opportunities of iPSC-assisted drug discovery with multi-disciplinary approaches from bioengineering to Omics technology. Despite technological challenges, iPSC-derived cytoarchitectures through interactions of diverse cell types mimic patients’ CNS and serve as a platform for therapeutic development and personalized precision medicine.

Advanced in vitro management of three-dimensional cell cultures and explanted tissue

Cytotherapy ◽  
2021 ◽  
Vol 23 (5) ◽  
pp. S145
Author(s):  
S. Kress ◽  
D. Egger ◽  
C. Kasper
Keyword(s):  
Cell Cultures ◽  
Three Dimensional ◽  
Dimensional Cell

A three-dimensional cell biology model of human hepatocellular carcinoma in vitro

Tumor Biology ◽  
2010 ◽  
Vol 32 (3) ◽  
pp. 469-479 ◽  
Author(s):  
Jianhua Tang ◽  
Jiefeng Cui ◽  
Rongxin Chen ◽  
Kun Guo ◽  
Xiaonan Kang ◽  
...  
Keyword(s):  
Hepatocellular Carcinoma ◽  
Cell Biology ◽  
Three Dimensional ◽  
Human Hepatocellular Carcinoma ◽  
Dimensional Cell
Sign in / Sign up

Export Citation Format

Share Document