Effects of Bisphosphonate on Long-Term Culture of Cartilage Allografts

2013 ◽  
Author(s):  
Yilu Zhou ◽  
Lauren Resutek ◽  
Liyun Wang ◽  
X. Lucas Lu
Keyword(s):  
Medial Meniscus ◽  
Biomechanical Properties ◽  
Culture Conditions ◽  
Protective Mechanisms ◽  
Post Traumatic ◽  
Biochemical Mechanisms ◽  
Altered Metabolism

Zoledronic acid (ZA), an FDA approved bisphosphonate (BP) medicine, is widely used for the treatment of osteoclast-related bone loss diseases [1]. Our previous study has found that systemic administration of ZA could dramatically suppress the development of post-traumatic osteoarthritis (PTOA) in the DMM (destabilization of the medial meniscus) mouse model, a model recapitulating the altered joint loading associated with PTOA [2]. This finding is consistent with a few similar studies using different animal models [3]. However, little is known about the cellular and biochemical mechanisms of BP mediated chondro-protection in PTOA pathogenesis. Studies have shown that PTOA often initiates from the apoptosis and altered metabolism of cartilage chondrocytes. In this study, we will investigate the direct effects of ZA on the metabolisms of chondrocytes using long-term in vitro culture of cartilage allografts. As one of the earliest responses of chondrocytes to mechanical stimulation, intracellular calcium ([Ca 2+] i) signaling is the upstream of numerous mechanotransduction pathways [4]. We hypothesize that the chondro-protective mechanisms of ZA could be represented by the characteristics of [Ca 2+] i signaling of in situ chondrocytes. Our specific aims were to: (i) compare the in situ spontaneous [Ca 2+] i responses of chondrocytes cultured in non-ZA and ZA supplemented environments, and (ii) compare the biomechanical properties of cartilage allografts under the two culture conditions.

scholarly journals Extracellular Matrix Determines Biomechanical Properties of Chondrospheres during Their Maturation In Vitro

Cartilage ◽  
2018 ◽  
Vol 11 (4) ◽  
pp. 521-531 ◽  
Author(s):  
Nikolai P. Omelyanenko ◽  
Pavel A. Karalkin ◽  
Elena A. Bulanova ◽  
Elizaveta V. Koudan ◽  
Vladislav A. Parfenov ◽  
...  
Keyword(s):  
Extracellular Matrix ◽  
Self Assembly ◽  
Biomechanical Properties ◽  
Culture Conditions ◽  
Secant Modulus ◽  
Structural Determinants ◽  
Maturation In Vitro

Objective Chondrospheres represent a variant of tissue spheroids biofabricated from chondrocytes. They are already being used in clinical trials for cartilage repair; however, their biomechanical properties have not been systematically investigated yet. The aim of our study was to characterize chondrospheres in long-term in vitro culture conditions for morphometric changes, biomechanical integrity, and their fusion and spreading kinetics. Results It has been demonstrated that the increase in chondrospheres secant modulus of elasticity is strongly associated with the synthesis and accumulation of extracellular matrix. Additionally, significant interplay has been found between biomechanical properties of tissue spheroids and their fusion kinetics in contrast to their spreading kinetics. Conclusions Extracellular matrix is one of the main structural determinants of chondrospheres biomechanical properties during chondrogenic maturation in vitro. The estimation of tissue spheroids’ physical behavior in vitro prior to operative treatment can be used to predict and potentially control fusogenic self-assembly process after implantation in vivo.

In Situ Calcium Signaling of Chondrocytes Under Non-Serum and Serum Culture

2013 ◽  
Author(s):  
Yilu Zhou ◽  
Lauren Resutek ◽  
Liyun Wang ◽  
X. Lucas Lu
Keyword(s):  
Calcium Signaling ◽  
Mechanical Stimulation ◽  
Biomechanical Properties ◽  
Cellular Level ◽  
Cartilage Explants ◽  
Serum Free ◽  
Better Than

Chemically defined serum-free medium has been shown to maintain the mechanical properties of cartilage allografts better than serum supplemented medium during long-term in vitro culture [1]. Little is known about this beneficial mechanism at a cellular level. Intracellular calcium ([Ca2+]i) signaling is one of the earliest responses in chondrocytes under mechanical stimulation [2]. It was recently found that calcium signaling is involved in the regulation of chondrocyte morphology changes and its short-term anabolic and catabolic responses under mechanical stimulation [3]. In this study we hypothesized that the beneficial mechanisms of serum-free culture could be indicated by the spatiotemporal features of [Ca2+]i signaling of chondrocytes in situ. We aimed to: (i) compare the in situ spontaneous [Ca2+]i responses of chondrocytes cultured in medium with and without serum; (ii) investigate the correlation between the [Ca2+]i responses of chondrocytes and the biomechanical properties of cartilage explants.

scholarly journals Activation of the receptor tyrosine kinase RET improves long-term hematopoietic stem cell outgrowth and potency

Blood ◽  
2020 ◽  
Vol 136 (22) ◽  
pp. 2535-2547 ◽  
Author(s):  
W. Grey ◽  
R. Chauhan ◽  
M. Piganeau ◽  
H. Huerga Encabo ◽  
M. Garcia-Albornoz ◽  
...  
Keyword(s):  
Intracellular Signaling ◽  
Culture Conditions ◽  
Cellular Growth ◽  
Great Promise ◽  
Bone Marrow Niche ◽  
Hematopoietic Stem ◽  
Intracellular Signaling Pathway ◽  
Wide Range

Abstract Expansion of human hematopoietic stem cells (HSCs) is a rapidly advancing field showing great promise for clinical applications. Recent evidence has implicated the nervous system and glial family ligands (GFLs) as potential drivers of hematopoietic survival and self-renewal in the bone marrow niche; how to apply this process to HSC maintenance and expansion has yet to be explored. We show a role for the GFL receptor, RET, at the cell surface of HSCs in mediating sustained cellular growth, resistance to stress, and improved cell survival throughout in vitro expansion. HSCs treated with the key RET ligand/coreceptor complex, glial-derived neurotrophic factor and its coreceptor, exhibit improved progenitor function at primary transplantation and improved long-term HSC function at secondary transplantation. Finally, we show that RET drives a multifaceted intracellular signaling pathway, including key signaling intermediates protein kinase B, extracellular signal-regulated kinase 1/2, NF-κB, and p53, responsible for a wide range of cellular and genetic responses that improve cell growth and survival under culture conditions.

Expression of the SMP antigen by oligodendrocytes in the developing avian central nervous system

Development ◽  
1989 ◽  
Vol 107 (4) ◽  
pp. 825-833
Author(s):  
P. Cameron-Curry ◽  
C. Dulac ◽  
N.M. Le Douarin
Keyword(s):  
Monoclonal Antibody ◽  
Schwann Cell ◽  
Basic Protein ◽  
Culture Conditions ◽  
Cell Marker ◽  
Cellular Expression ◽  
Early Expression ◽  
Different Parts

Expression of the avian antigen SMP (Schwann cell Myelin Protein, Mr 75-80000), first characterized in the PNS with a monoclonal antibody as an early and strictly specific Schwann cell marker, was further studied in the CNS. Comparing SMP immunoreactive areas in the different parts of the CNS with those expressing the Myelin Basic Protein (MBP), we showed a strict colocalisation of both phenotypes. In vitro, MBP+ oligodendrocytes express the surface antigen SMP as well. SMP cellular expression was followed in situ and in culture using nervous tissues from embryos at different stages. We were thus able to detect an early expression of this marker by oligodendroblasts before the first appearance of MBP immunoreactivity. We have also identified a subpopulation of SMP+/MBP- and SMP+/GC- cells, which persists under our culture conditions as precursors remaining in an immature state.

Structure of vegetative organs in essential oil rose under standard culture conditions and long-term conservation in vitro

2021 ◽  
pp. 185-190
Author(s):  
I.V. Mitrofanova ◽  
V.A. Brailko ◽  
N.P. Lesnikova-Sedoshenko ◽  
N.N. Ivanova ◽  
O.V. Mitrofanova
Keyword(s):  
Essential Oil ◽  
Culture Conditions ◽  
Vegetative Organs ◽  
Standard Culture

scholarly journals Cycle initiation and colony formation in culture by murine marrow cells with long-term reconstituting potential in vivo

Blood ◽  
1996 ◽  
Vol 88 (11) ◽  
pp. 4149-4158 ◽  
Author(s):  
M Trevisan ◽  
XQ Yan ◽  
NN Iscove
Keyword(s):  
Colony Formation ◽  
Interleukin 1 ◽  
Methyl Cellulose ◽  
Culture Conditions ◽  
Liquid Suspension ◽  
Almost All ◽  
Marrow Cells

Abstract This investigation was directed at separating long-term reconstituting (LTR) stem cells in normal murine marrow from hematopoietic precursors detectable in short-term assays in vitro and in vivo, and then at determining whether purified LTR cells could themselves form colonies in culture. To do so, it was first necessary to identify culture conditions that would induce their growth while preserving their long- term reconstituting capacity. Marrow was cultured with various cytokines in liquid suspension for 4 days, after which the surviving LTR activity was quantitated in a competitive in vivo assay. Activity was preserved near input levels with combined murine c-kit ligand (KL), interleukin-1 (IL-1), IL-6, and IL-11. When the cultures also included tritiated or unlabeled thymidine, LTR potential was eliminated, indicating that essentially all LTR cells were induced into cell cycle with these cytokines. To purify them, marrow was sorted on the basis of Ly6A expression and Rhodamine 123 retention. The Ly6AhiRh123ls fraction contained 85% of total recovered LTR activity but only 1% of the recovered cells measured by multilineage colony formation in spleens or in vitro. This fraction was cultured in methyl cellulose with KL, IL-1, IL-6, and IL-11 for 4 to 6 days, after which colonies were isolated and injected into mice. High levels of permanent reconstitution were achievable in sublethally irradiated W41/W41 mice after the injection of a single reconstituting unit, and limiting dilution analysis estimated the frequency of multilineage LTR at 1 in 11,200 unpurified adult marrow cells. In either lethally irradiated normal or sublethally irradiated W41/W41 mice, 1-year lymphomyeloid reconstitutions were obtained from 1 in 65 to 84 colonies of 2 to 16 dispersed cells, but not from larger colonies or those with clumped cells. The results establish that resting marrow LTR cells can be separated from almost all of the more advanced clonogenic cells that are still pluripotential, can be induced to cycle in culture by defined cytokines with preservation of their reconstituting potential, and can be manipulated and assayed efficiently at single-cell and colony levels.

scholarly journals Human macrophage maturation and heterogeneity: analysis with a newly generated set of monoclonal antibodies to differentiation antigens

Blood ◽  
1986 ◽  
Vol 67 (5) ◽  
pp. 1257-1264 ◽  
Author(s):  
R Andreesen ◽  
KJ Bross ◽  
J Osterholz ◽  
F Emmrich
Keyword(s):  
Monoclonal Antibodies ◽  
Cell Surface ◽  
Cell Lineage ◽  
Culture Conditions ◽  
Human Macrophage ◽  
Blood Monocytes ◽  
Differentiation Antigens

We have analyzed the expression of late differentiation antigens during terminal in vitro maturation of human macrophages (M phi) from blood monocytes (MO) in comparison to their distribution among mature M phi residing in various tissue sites. By immunizing mice with M phi derived from blood MO by culture on hydrophobic Teflon foils, monoclonal antibodies (mAbs) were developed (MAX.1, MAX.2, MAX.3, MAX.11) that reacted with lineage-restricted differentiation antigens. These antigens were expressed exclusively on M phi or were markedly increased after in vitro differentiation. The only overlap to another hemopoietic cell lineage was observed with MAX.3, which is shared by platelets and megakaryocytes. In the course of M phi maturation in vitro, the MAX.1 and MAX.3 antigens are detected within the cytoplasm two days before they appear on the cell surface. In contrast, the MAX.11 antigen is expressed simultaneously in the cytoplasm and at the cell surface, is found in varying degrees on a minor portion of blood MO and U937 cells, and is expressed rapidly at high density during early M phi differentiation in vitro. Among conventional mAbs that do not react with MO we found those against the transferrin (TF)-receptor, the BA-2, and the PCA1 antigen to label M phi. M phi matured in vivo and isolated from body fluids were positive with some but not all MAX mAbs. Distinctive patterns were observed with pulmonary M phi, exudate M phi from pleural and peritoneal effusions, synovial fluids, and early lactation milk. M phi from the alveolar space, for example, constantly expressed the MAX.2 antigen but not the MAX.3 antigen. Pleural effusion M phi, however, did not react with the MAX.1 mAb, but in most cases, it did react with the MAX.3 mAb. The detection of novel differentiation antigens, all expressed on monocyte-derived M phi but differently expressed on site-specific M phi in situ, underlines the remarkable heterogeneity among human M phi. The expression of these antigens is flexible because those MAX antigens that were not expressed in situ could be induced if cells from distinct tissue sites were cultured in vitro for several days. MAX mAbs may be of potential value to study both the sequential stages of maturation within the M phi lineage as well as differential developments induced by various culture conditions in parallel to environmental factors in vivo.

scholarly journals Stromal cells in myeloid and lymphoid long-term bone marrow cultures can support multiple hemopoietic lineages and modulate their production of hemopoietic growth factors

Blood ◽  
1986 ◽  
Vol 68 (6) ◽  
pp. 1348-1354 ◽  
Author(s):  
A Johnson ◽  
K Dorshkind
Keyword(s):  
Bone Marrow ◽  
Stromal Cells ◽  
Stromal Cell ◽  
Culture Conditions ◽  
B Lymphopoiesis ◽  
Bone Marrow Cultures ◽  
Factor Secretion ◽  
Marrow Cultures

Abstract Hemopoiesis in long-term bone marrow cultures (LTBMC) is dependent on adherent stromal cells that form an in vitro hemopoietic microenvironment. Myeloid bone marrow cultures (MBMC) are optimal for myelopoiesis, while lymphoid bone marrow cultures (LBMC) only support B lymphopoiesis. The experiments reported here have made a comparative analysis of the two cultures to determine whether the stromal cells that establish in vitro are restricted to the support of myelopoiesis or lymphopoiesis, respectively, and to examine how the different culture conditions affect stromal cell physiology. In order to facilitate this analysis, purified populations of MBMC and LBMC stroma were prepared by treating the LTBMC with the antibiotic mycophenolic acid; this results in the elimination of hemopoietic cells while retaining purified populations of functional stroma. Stromal cell cultures prepared and maintained under MBMC conditions secreted myeloid growth factors that stimulated the growth of granulocyte-macrophage colonies, while no such activity was detected from purified LBMC stromal cultures. However, this was not due to the inability of LBMC stroma to mediate this function. Transfer of LBMC stromal cultures to MBMC conditions resulted in an induction of myeloid growth factor secretion. When seeded under these conditions with stromal cell- depleted populations of hemopoietic cells, obtained by passing marrow through nylon wool columns, the LBMC stromal cells could support long- term myelopoiesis. Conversely, transfer of MBMC stroma to LBMC conditions resulted in a cessation of myeloid growth factor secretion; on seeding these cultures with nylon wool-passed marrow, B lymphopoiesis, but not myelopoiesis, initiated. These findings indicate that the stroma in the different LTBMC are not restricted in their hemopoietic support capacity but are sensitive to culture conditions in a manner that may affect the type of microenvironment formed.

In situ generation of human brain organoids on a micropillar array

Lab on a Chip ◽  
2017 ◽  
Vol 17 (17) ◽  
pp. 2941-2950 ◽  
Author(s):  
Yujuan Zhu ◽  
Li Wang ◽  
Hao Yu ◽  
Fangchao Yin ◽  
Yaqing Wang ◽  
...  
Keyword(s):  
Stem Cells ◽  
Human Brain ◽  
Pluripotent Stem Cells ◽  
3D Culture ◽  
In Situ Generation ◽  
Induced Pluripotent ◽  
High Throughput Manner

We present a simple and high throughput manner to generate brain organoids in situ from human induced pluripotent stem cells on micropillar arrays and to investigate long-term brain organogenesis in 3D culture in vitro.

Adaptation to calcium deprivation in the rat: effects of parathyroidectomy.

1977 ◽  
Vol 232 (3) ◽  
pp. E336
Author(s):  
J T Pento ◽  
L C Waite ◽  
P J Tracy ◽  
A D Kenny
Keyword(s):  
Adaptive Response ◽  
Calcium Transport ◽  
Parathyroid Tissue ◽  
Adaptation Period ◽  
Short Term ◽  
High Calcium

The role of parathyroid hormone (PTH) in the adaptive response in gut calcium transport to calcium deprivation has been studied in the rat using both the in vitro everted duodenal sac and the in situ ligated duodenal segment technique. Intact or parathyroidectomized (PTX) young rats were placed on a low calcium (0.01%) diet for 7-, 14-, or 21-day adaptation periods and compared with control rats maintained on a high calcium (1.5%) diet. Prior PTX (3 days before the start of the adaptation period) abolished the adaptive response (enhanced calcium transport) induced by calcium deprivation for a 7-day adaptation period, but did not abolish a response after a 21-day period. A 14-day adaptation period gave equivocal results. It is concluded that PTH appears to be necessary for short-term (7-day) adaptation, but not for long-term (21-day) adaptation to calcium deprivation. However, if accessory parathyroid tissue is present, the data could be interpreted differently: the essentiality of PTH for the adaptive response might be independent of the length of the adaptation period. The data also contribute to a possible resolution of the controversy concerning the involvement of PTH in the regulation of intestinal calcium transport in the rat.

Sign in / Sign up

Export Citation Format

Share Document