scholarly journals The potentials and pitfalls of a human cervical organoid model including Langerhans cells

2019 ◽  
Author(s):  
Robert Jackson ◽  
Jordan D Lukacs ◽  
Ingeborg Zehbe
Keyword(s):  
Langerhans Cells ◽  
Three Dimensional ◽  
Myeloid Cell ◽  
Petri Dish ◽  
Cervical Tissue ◽  
Myeloid Cell Line ◽  
Versatile Tool ◽  
Research Questions ◽  
Dimensional Cell ◽  
Detailed Protocol

AbstractThree-dimensional cell culturing to capture a life-like experimental environment has become a versatile tool for basic and clinical research. Mucosal and skin tissues can be grown as “organoids” in a petri dish and serve a wide variety of research questions. Here, we report our experience with human cervical organoids which could also include an immune component, e.g. Langerhans cells. We employ commercially available human cervical keratinocytes and fibroblasts as well as a myeloid cell line matured and purified into langerin-positive Langerhans cells. These are then seeded on a layer of keratinocytes with underlying dermal equivalent. Using about 10-fold more than the reported number in healthy cervical tissue (1–3%), we obtain differentiated cervical epithelium after 14 days with ~ 1% being Langerhans cells. We provide a detailed protocol for interested researchers to apply the described “aseptic” organoid model for all sorts of investigations—with or without Langerhans cells.

scholarly journals The Potentials and Pitfalls of a Human Cervical Organoid Model Including Langerhans Cells

Viruses ◽  
2020 ◽  
Vol 12 (12) ◽  
pp. 1375
Author(s):  
Robert Jackson ◽  
Jordan D. Lukacs ◽  
Ingeborg Zehbe
Keyword(s):  
Langerhans Cells ◽  
Three Dimensional ◽  
Myeloid Cell ◽  
Petri Dish ◽  
Cervical Tissue ◽  
Myeloid Cell Line ◽  
Versatile Tool ◽  
Research Questions ◽  
Dimensional Cell ◽  
Detailed Protocol

Three-dimensional cell culturing to capture a life-like experimental environment has become a versatile tool for basic and clinical research. Mucosal and skin tissues can be grown as “organoids” in a petri dish and serve a wide variety of research questions. Here, we report our experience with human cervical organoids which could also include an immune component, e.g., Langerhans cells. We employ commercially available human cervical keratinocytes and fibroblasts as well as a myeloid cell line matured and purified into langerin-positive Langerhans cells. These are then seeded on a layer of keratinocytes with underlying dermal equivalent. Using about 10-fold more than the reported number in healthy cervical tissue (1–3%), we obtain differentiated cervical epithelium after 14 days with ~1% being Langerhans cells. We provide a detailed protocol for interested researchers to apply the described “aseptic” organoid model for all sorts of investigations—with or without Langerhans cells.

scholarly journals Non-contact three-dimensional cell cluster formation on demand in open dishware using focused surface acoustic waves through a couplant layer

2021 ◽  
Author(s):  
Jiyang Mei ◽  
Aditya Vasan ◽  
Uri Magaram ◽  
Kenjiro Takemura ◽  
Sreekanth Chalasani ◽  
...  
Keyword(s):  
Acoustic Waves ◽  
Drug Testing ◽  
Cluster Formation ◽  
Surface Acoustic Waves ◽  
Three Dimensional ◽  
Petri Dish ◽  
Hek293 Cells ◽  
Sound Waves ◽  
The Media ◽  
Dimensional Cell

Three-dimensional cell agglomerates are broadly useful in tissue engineering and drug testing. We report a well-free method to form large (1-mm) multicellular clusters using 100-MHz surface acoustic waves (SAW) without direct contact with the media or cells. A fluid couplant is used to transformthe SAW into acoustic streaming in the cell-laden media held in a petri dish. The couplant transmits longitudinal sound waves, forming a Lamb wave in the petri dish that, in turn, produces longitudinal sound in the media. Due to recirculation, human embryonic kidney (HEK293) cells in the dish are carried to the center of the coupling location, forming a cluster in less than 10 min. A few minutes later, these clusters may then be translated and merged to form large agglomerations, and even repeatedly folded to produce a roughly spherical shape of over 1 mm in diameter for incubation—without damaging the existing intercellular bonds. Calcium ion signaling through these clusters and confocal images of multiprotein junctional complexes suggest a continuous tissue construct: intercellular communication. They may be formed at will, and the method is feasibly useful for formation of numerous agglomerates in a single petri dish.

scholarly journals Can keratin scaffolds be used for creating three-dimensional cell cultures?

Open Medicine ◽  
2020 ◽  
Vol 15 (1) ◽  
pp. 249-253
Author(s):  
Marta Bochynska-Czyz ◽  
Patrycja Redkiewicz ◽  
Hanna Kozlowska ◽  
Joanna Matalinska ◽  
Marek Konop ◽  
...  
Keyword(s):  
Cell Cultures ◽  
Chemical Activation ◽  
Three Dimensional ◽  
Enzymatic Digestion ◽  
Pepsin Digestion ◽  
Cell Culturing ◽  
3D Cell Cultures ◽  
Associated Proteins ◽  
Positive Effect ◽  
Dimensional Cell

AbstractThree-dimensional (3D) cell cultures were created with the use of fur keratin associated proteins (F-KAPs) as scaffolds. The procedure of preparation F-KAP involves combinations of chemical activation and enzymatic digestion. The best result in porosity and heterogeneity of F-KAP surface was received during pepsin digestion. The F-KAP had a stable structure, no changes were observed after heat treatment, shaking and washing. The 0.15-0.5 mm fraction had positive effect for formation of 3D scaffolds and cell culturing. Living rat mesenchymal cells on the F-KAP with no abnormal morphology were observed by SEM during 32 days of cell culturing.

Three-dimensional cell-adhesive matrix of silk cocoon derived carbon fiber assembled with iron-porphyrin for monitoring cell released signal molecules

2021 ◽  
Vol 334 ◽  
pp. 129594
Author(s):  
Fang Xin Hu ◽  
Xiaoli Xie ◽  
Dongping Wang ◽  
Hong Bin Yang ◽  
Yu Gu ◽  
...  
Keyword(s):  
Carbon Fiber ◽  
Three Dimensional ◽  
Iron Porphyrin ◽  
Signal Molecules ◽  
Silk Cocoon ◽  
Adhesive Matrix ◽  
Cell Adhesive ◽  
Dimensional Cell

scholarly journals Propagation‐based Phase Contrast Computed Tomography as a Suitable Tool for the Characterization of Spatial Three‐Dimensional Cell Distribution in Biomaterials

2021 ◽  
Author(s):  
D.C. Florian Wieland ◽  
Simone Krueger ◽  
Julian Moosmann ◽  
Thomas Distler ◽  
Alina Weizel ◽  
...  
Keyword(s):  
Computed Tomography ◽  
Phase Contrast ◽  
Three Dimensional ◽  
Cell Distribution ◽  
Suitable Tool ◽  
Dimensional Cell

scholarly journals Factors to consider when interrogating 3D culture models with plate readers or automated microscopes

2021 ◽  
Author(s):  
Terry Riss ◽  
O. Joseph Trask
Keyword(s):  
Cell Culture ◽  
Three Dimensional ◽  
3D Culture ◽  
Fluorescent Imaging ◽  
Culture Models ◽  
Assay Methods ◽  
Diffusion Rates ◽  
Cell Culture Models ◽  
Dimensional Cell ◽  
Cells Cultured

AbstractAlong with the increased use of more physiologically relevant three-dimensional cell culture models comes the responsibility of researchers to validate new assay methods that measure events in structures that are physically larger and more complex compared to monolayers of cells. It should not be assumed that assays designed using monolayers of cells will work for cells cultured as larger three-dimensional masses. The size and barriers for penetration of molecules through the layers of cells result in a different microenvironment for the cells in the outer layer compared to the center of three-dimensional structures. Diffusion rates for nutrients and oxygen may limit metabolic activity which is often measured as a marker for cell viability. For assays that lyse cells, the penetration of reagents to achieve uniform cell lysis must be considered. For live cell fluorescent imaging assays, the diffusion of fluorescent probes and penetration of photons of light for probe excitation and fluorescent emission must be considered. This review will provide an overview of factors to consider when implementing assays to interrogate three dimensional cell culture models.

scholarly journals HiChIP and Hi-C Protocol Optimized for Primary Murine T Cells

2021 ◽  
Vol 4 (3) ◽  
pp. 49
Author(s):  
Tomas Zelenka ◽  
Charalampos Spilianakis
Keyword(s):  
T Cells ◽  
Long Range ◽  
Three Dimensional ◽  
Cell Types ◽  
Range Interaction ◽  
Chromatin Interactions ◽  
Optimized Protocol ◽  
Long Range Interaction ◽  
Chromatin Compactness ◽  
Detailed Protocol

The functional implications of the three-dimensional genome organization are becoming increasingly recognized. The Hi-C and HiChIP research approaches belong among the most popular choices for probing long-range chromatin interactions. A few methodical protocols have been published so far, yet their reproducibility and efficiency may vary. Most importantly, the high frequency of the dangling ends may dramatically affect the number of usable reads mapped to valid interaction pairs. Additionally, more obstacles arise from the chromatin compactness of certain investigated cell types, such as primary T cells, which due to their small and compact nuclei, impede limitations for their use in various genomic approaches. Here we systematically optimized all the major steps of the HiChIP protocol in T cells. As a result, we reduced the number of dangling ends to nearly zero and increased the proportion of long-range interaction pairs. Moreover, using three different mouse genotypes and multiple biological replicates, we demonstrated the high reproducibility of the optimized protocol. Although our primary goal was to optimize HiChIP, we also successfully applied the optimized steps to Hi-C, given their significant protocol overlap. Overall, we describe the rationale behind every optimization step, followed by a detailed protocol for both HiChIP and Hi-C experiments.

TRANSFORMATION BY V-ERB-B - INDUCTION OF APOPTOSIS IS ABROGATED IN A MYELOID CELL-LINE

1995 ◽  
Author(s):  
A DOLNIKOV ◽  
Y SHOUNAN ◽  
NJ HAWKINS ◽  
RL WARD ◽  
G SYMONDS
Keyword(s):  
Cell Line ◽  
Myeloid Cell ◽  
Myeloid Cell Line ◽  
Induction Of Apoptosis
Sign in / Sign up

Export Citation Format

Share Document