Comparison of 3D culture methods on human HepG2 cells

Background: The effects of lipopolysaccharide (LPS) on cell proliferation and osteogenic potential (OP) of MSCs have been frequently studied. Objective: to compare the effects of LPS on periodontal-ligament-derived mesenchymal stem cells (PDLSCs) in monolayer and 3D culture. Methods: The PDLSCs were colorimetrically assessed for proliferation and osteogenic potential (OP) after LPS treatment. The 3D cells were manually prepared by scratching and allowing them to clump up. The clumps (C-MSCs) were treated with LPS and assessed for Adenosine triphosphate (ATP) and OP. Raman spectroscopy was used to analyze calcium salts, DNA, and proline/hydroxyproline. Multiplexed ELISA was performed to assess LPS induced local inflammation. Results: The proliferation of PDLSCs decreased with LPS. On Day 28, LPS-treated cells showed a reduction in their OP. C-MSCs with LPS did not show a decrease in ATP production. Principal bands identified in Raman analysis were the P–O bond at 960 cm−1 of the mineral component, 785 cm−1, and 855 cm−1 showing qualitative changes in OP, proliferation, and proline/hydroxyproline content, respectively. ELISA confirmed increased levels of IL-6 and IL-8 but with the absence of TNF-α and IL-1β secretion. Conclusions: These observations demonstrate that C-MSCs are more resistant to the effects of LPS than cells in monolayer cell culture. Though LPS stimulation of C-MSCs creates an early pro-inflammatory milieu by secreting IL-6 and IL-8, PDLSCs possess inactivated TNF promoter and an ineffective caspase-1 activating process.

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Human Adipose-Derived Stromal/Stem Cell Culture and Analysis Methods for Adipose Tissue Modeling In Vitro: A Systematic Review

Cells ◽

10.3390/cells10061378 ◽

2021 ◽

Vol 10 (6) ◽

pp. 1378

Author(s):

Peyton Gibler ◽

Jeffrey Gimble ◽

Katie Hamel ◽

Emma Rogers ◽

Michael Henderson ◽

...

Keyword(s):

Systematic Review ◽

Adipose Tissue ◽

Drug Discovery ◽

3D Culture ◽

Human Adipose Tissue ◽

Culture Methods ◽

Adipose Tissue Engineering ◽

The Impact

Human adipose-derived stromal/stem cells (hASC) are widely used for in vitro modeling of physiologically relevant human adipose tissue. These models are useful for the development of tissue constructs for soft tissue regeneration and 3-dimensional (3D) microphysiological systems (MPS) for drug discovery. In this systematic review, we report on the current state of hASC culture and assessment methods for adipose tissue engineering using 3D MPS. Our search efforts resulted in the identification of 184 independent records, of which 27 were determined to be most relevant to the goals of the present review. Our results demonstrate a lack of consensus on methods for hASC culture and assessment for the production of physiologically relevant in vitro models of human adipose tissue. Few studies have assessed the impact of different 3D culture conditions on hASC adipogenesis. Additionally, there has been a limited use of assays for characterizing the functionality of adipose tissue in vitro. Results from this study suggest the need for more standardized culture methods and further analysis on in vitro tissue functionality. These will be necessary to validate the utility of 3D MPS as an in vitro model to reduce, refine, and replace in vivo experiments in the drug discovery regulatory process.

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Evaluation of Spheroid 3D Culture Methods to Study a Pancreatic Neuroendocrine Neoplasm Cell Line

Frontiers in Endocrinology ◽

10.3389/fendo.2019.00682 ◽

2019 ◽

Vol 10 ◽

Cited By ~ 5

Author(s):

Giulia Bresciani ◽

Leo J. Hofland ◽

Fadime Dogan ◽

Georgios Giamas ◽

Teresa Gagliano ◽

...

Keyword(s):

Cell Line ◽

3D Culture ◽

Neuroendocrine Neoplasm ◽

Culture Methods ◽

Pancreatic Neuroendocrine Neoplasm

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Quercetin potentializes the respective cytotoxic activity of gemcitabine or doxorubicin on 3D culture of AsPC-1 or HepG2 cells, through the inhibition of HIF-1α and MDR1

PLoS ONE ◽

10.1371/journal.pone.0240676 ◽

2020 ◽

Vol 15 (10) ◽

pp. e0240676

Author(s):

Sarah Hassan ◽

Jean Peluso ◽

Sandra Chalhoub ◽

Ysia Idoux Gillet ◽

Nadia Benkirane-Jessel ◽

...

Keyword(s):

Cytotoxic Activity ◽

Hepg2 Cells ◽

3D Culture

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Long-term 3D culture of the SCC4 cell line using three different culture methods and initial seeding densities

Journal of Cellular Biotechnology ◽

10.3233/jcb-179005 ◽

2017 ◽

Vol 3 (1) ◽

pp. 41-50 ◽

Cited By ~ 5

Author(s):

Valeh Rustamov ◽

Rüdiger Rudolf ◽

Vugar Yagublu ◽

Hella-Monika Kuhn ◽

Mario Vitacolonna ◽

...

Keyword(s):

Cell Line ◽

3D Culture ◽

Culture Methods ◽

Scc4 Cell ◽

Initial Seeding

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Neurosphere Based Differentiation of Human iPSC Improves Astrocyte Differentiation

Stem Cells International ◽

10.1155/2016/4937689 ◽

2016 ◽

Vol 2016 ◽

pp. 1-15 ◽

Cited By ~ 23

Author(s):

Shuling Zhou ◽

Karolina Szczesna ◽

Anna Ochalek ◽

Julianna Kobolák ◽

Eszter Varga ◽

...

Keyword(s):

Neural Progenitor Cells ◽

Three Dimensional ◽

3D Culture ◽

Neural Progenitor Cell ◽

Neural Progenitor ◽

Culture Methods ◽

Differentiation Potential ◽

Differentiated Cells ◽

Astrocyte Differentiation ◽

Induced Pluripotent

Neural progenitor cells (NPCs) derived from human induced pluripotent stem cells (iPSCs) are traditionally maintained and proliferated utilizing two-dimensional (2D) adherent monolayer culture systems. However, NPCs cultured using this system hardly reflect the intrinsic spatial development of brain tissue. In this study, we determined that culturing iPSC-derived NPCs as three-dimensional (3D) floating neurospheres resulted in increased expression of the neural progenitor cell (NPC) markers,PAX6andNESTIN. Expansion of NPCs in 3D culture methods also resulted in a more homogenous PAX6 expression when compared to 2D culture methods. Furthermore, the 3D propagation method for NPCs resulted in a significant higher expression of the astrocyte markers GFAPandaquaporin 4(AQP4) in the differentiated cells. Thus, our 3D propagation method could constitute a useful tool to promote NPC homogeneity and also to increase the differentiation potential of iPSC towards astrocytes.

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An extracellular matrix-based culture system for generation of human pluripotent stem cell derived-hepatocytes

Current Stem Cell Research & Therapy ◽

10.2174/1574888x16666201228144834 ◽

2020 ◽

Vol 16 ◽

Author(s):

Mehdi Forouzesh ◽

Mojgan Hosseini ◽

Mehran Ataei ◽

Maryam Farzaneh ◽

Seyed Esmaeil Khoshnam

Keyword(s):

Stem Cells ◽

Extracellular Matrix ◽

Pluripotent Stem Cells ◽

Liver Diseases ◽

Therapeutic Potential ◽

Embryonic Stem ◽

3D Culture ◽

Hepatic Differentiation ◽

Human Escs ◽

Culture Methods

: Liver disease (hepatic disease) adversely affects the normal function of the liver and causes liver problems. Druginduced liver injury (DILI) can be predicted by primary human hepatocytes. However, the sources of hepatocytes for largescale drug toxicity screening are limited. To solve this problem, pluripotent stem cells (PSCs), mesenchymal stem cells (MSCs), and hepatic stem cells (HSCs) have emerged as attractive cell sources for cell-based therapies. Human PSCs including embryonic stem cells (ESCs) and induced pluripotent stem cells (iPSCs) have the ability to undergo self-renewal and to differentiate into lineages of ectoderm, mesoderm, and endoderm. Human PSC can be used for generation of hepatocytes to facilitate the development of novel drugs for treatment of severe liver diseases. The therapeutic potential of PSC-derived hepatocytes for liver failure have been identified to enhance the development of chemically defined and xenogenic-free 3D culture methods. To date, several hepatic differentiation strategies and various extracellular matrix (ECM) components have been employed to produce hepatocytes or hepatic-like cells (HLCs) in vitro. In this review, we focused on the potential of Matrigel, collagen type 1, RoGel, and laminin as ECM on the differentiation and function of hESC- and hiPSC-derived hepatocytes. The hepatic differentiation of human ESCs and iPSCs would offer an ideal tool for cell therapy and liver diseases.

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A novel chemotherapeutic sensitivity-testing system based on collagen gel droplet embedded 3D–culture methods for hepatocellular carcinoma

BMC Cancer ◽

10.1186/s12885-017-3706-6 ◽

2017 ◽

Vol 17 (1) ◽

Cited By ~ 16

Author(s):

Jun Hou ◽

Zhixian Hong ◽

Fan Feng ◽

Yantao Chai ◽

Yunkai Zhang ◽

...

Keyword(s):

Hepatocellular Carcinoma ◽

3D Culture ◽

Collagen Gel ◽

Testing System ◽

Culture Methods ◽

Sensitivity Testing ◽

Chemotherapeutic Sensitivity

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3D Stem Cell Culture

Cells ◽

10.3390/cells9102178 ◽

2020 ◽

Vol 9 (10) ◽

pp. 2178

Author(s):

Joni H. Ylostalo

Keyword(s):

Stem Cells ◽

Stem Cell ◽

Cell Biology ◽

3D Culture ◽

Basic Research ◽

The Body ◽

Culture Methods ◽

Culture Techniques ◽

3D Cultures

Much interest has been directed towards stem cells, both in basic and translational research, to understand basic stem cell biology and to develop new therapies for many disorders. In general, stem cells can be cultured with relative ease, however, most common culture methods for stem cells employ 2D techniques using plastic. These cultures do not well represent the stem cell niches in the body, which are delicate microenvironments composed of not only stem cells, but also supporting stromal cells, extracellular matrix, and growth factors. Therefore, researchers and clinicians have been seeking optimal stem cell preparations for basic research and clinical applications, and these might be attainable through 3D culture of stem cells. The 3D cultures recapitulate the in vivo cell-to-cell and cell-to-matrix interactions more effectively, and the cells in 3D cultures exhibit many unique and desirable characteristics. The culture of stem cells in 3D may employ various matrices or scaffolds, in addition to the cells, to support the complex structures. The goal of this Special Issue is to bring together recent research on 3D cultures of various stem cells to increase the basic understanding of stem cells and culture techniques, and also highlight stem cell preparations for possible novel therapeutic applications.

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