Microtechnology-based three-dimensional spheroid formation

10.2741/e594 ◽  
2013 ◽  
Vol E5 (1) ◽  
pp. 37-48 ◽  
Author(s):  
Hiroki Ota
Keyword(s):  
Three Dimensional ◽  
Spheroid Formation

scholarly journals Dexamethasone Inhibits Spheroid Formation of Thyroid Cancer Cells Exposed to Simulated Microgravity

Cells ◽  
2020 ◽  
Vol 9 (2) ◽  
pp. 367 ◽  
Author(s):  
Melnik ◽  
Sahana ◽  
Corydon ◽  
Kopp ◽  
Nassef ◽  
...  
Keyword(s):  
Thyroid Cancer ◽  
Cancer Cells ◽  
Cancer Progression ◽  
Cancer Biology ◽  
Expression Patterns ◽  
Three Dimensional ◽  
Signal Pathways ◽  
Dependent Manner ◽  
Spheroid Formation ◽  
Mesenchymal Transition

Detachment and the formation of spheroids under microgravity conditions can be observed with various types of intrinsically adherent human cells. In particular, for cancer cells this process mimics metastasis and may provide insights into cancer biology and progression that can be used to identify new drug/target combinations for future therapies. By using the synthetic glucocorticoid dexamethasone (DEX), we were able to suppress spheroid formation in a culture of follicular thyroid cancer (FTC)-133 cells that were exposed to altered gravity conditions on a random positioning machine. DEX inhibited the growth of three-dimensional cell aggregates in a dose-dependent manner. In the first approach, we analyzed the expression of several factors that are known to be involved in key processes of cancer progression such as autocrine signaling, proliferation, epithelial–mesenchymal transition, and anoikis. Wnt/β-catenin signaling and expression patterns of important genes in cancer cell growth and survival, which were further suggested to play a role in three-dimensional aggregation, such as NFKB2, VEGFA, CTGF, CAV1, BCL2(L1), or SNAI1, were clearly affected by DEX. Our data suggest the presence of a more complex regulation network of tumor spheroid formation involving additional signal pathways or individual key players that are also influenced by DEX.

Study on Pipetting Motion Optimization of Automatic Spheroid Culture System for Spheroid Formation

2021 ◽  
Vol 33 (1) ◽  
pp. 78-87
Author(s):  
Takeshi Shimoto ◽  
Chihiro Teshima ◽  
Toshiki Watanabe ◽  
Xiu-Ying Zhang ◽  
Atsushi Ishikawa ◽  
...  
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Culture System ◽  
Three Dimensional ◽  
Spheroid Formation ◽  
Spheroid Culture ◽  
Motion Optimization ◽  
Passage Number ◽  
Dimensional Cell ◽  
Number Of Cells

This research group has established a technology for producing a three-dimensional cell constructed using only the cell itself. This technology uses a property in which the spheroids fuse with each other. We developed a system that automates the spheroid production process to obtain reproducible spheroids and suppress variation factors that occur from human operation. However, it has become clear that the dispersion occurs in the diameter depending on the number of cells of the spheroid even if the cells are handled in the same manner. The purpose of this research is to examine an appropriate pipetting motion in accordance with the number of cells of the spheroid to be produced. Rabbit mesenchymal stem cells (rMSCs) are used as the objects. The number of cells was set to 2×104, 3×104, and 4×104 cells/well, and the passage number as 7. The appearance of spheroids cultured using the motion programmed in accordance with each number of cells was observed every 24 hours for 5 days after seeding. The results of the analysis indicate that the optimum motion in each number of cells has been successfully specified, and reproducible spheroids have been successfully produced.

scholarly journals Mesenchymal stem cell spheroids: potential cell materials for cell therapy

STEMedicine ◽  
2020 ◽  
Vol 2 (5) ◽  
pp. e67
Author(s):  
Zhongjuan Xu ◽  
Xingzhi Liu ◽  
Yu Wei ◽  
Zhe Zhao ◽  
Junjun Cao ◽  
...  
Keyword(s):  
Regenerative Medicine ◽  
Tissue Injury ◽  
Three Dimensional ◽  
3D Culture ◽  
Research Field ◽  
Three Dimensions ◽  
Spheroid Formation ◽  
Culture Methods ◽  
Differentiation Potential

Mesenchymal stromal/stem cells (MSCs) have been applied in clinical trials with an increasing number in recent years. MSCs showed their great potentials in regenerative medicine for their extensive sources, multilineage differentiation potential, low immunogenicity and self-renewal ability. However, the clinical application of MSCs still confronts many challenges including the requirement of large quantity of cells, low survival ability in vivo and the loss of main original characteristics due to two-dimensional (2D) culture although it is beneficial to cells fast expansion. Three-dimensional (3D) culture artificially creates an environment that permits cells to grow or interact with their surroundings in all three dimensions. Therefore, 3D culture was widely regarded as a more preferable and closer physiological microenvironment for cells growth. Recently, many different 3D spheroid culture methods have been developed to optimize MSCs biological characteristics to meet the demand of regenerative medicine. In this review, we comprehensively discussed the merits and demerits of different spheroid formation methods, expounded the mechanisms of spheroid formation and its microenvironment, and illustrated their optimized biological functions and the pre-clinical applications in various tissue injury and regeneration. In the end, we prospected the trends of this research field and proposed the key problems needed to be solved in the future.

scholarly journals Serum-Free Culture System for Spontaneous Human Mesenchymal Stem Cell Spheroid Formation

2019 ◽  
Vol 2019 ◽  
pp. 1-12 ◽  
Author(s):  
Guoyi Dong ◽  
Shengpeng Wang ◽  
Yuping Ge ◽  
Qiuting Deng ◽  
Qi Cao ◽  
...  
Keyword(s):  
Clinical Research ◽  
Expression Profiles ◽  
Three Dimensional ◽  
3D Culture ◽  
Human Mesenchymal Stem Cells ◽  
Human Mesenchymal Stem Cell ◽  
Spheroid Formation ◽  
Culture Methods ◽  
Serum Replacement ◽  
Knockout Serum Replacement

Human mesenchymal stem cells (hMSCs) are widely used in clinical research because of their multipotential, immunomodulatory, and reparative properties. Previous studies determined that hMSC spheroids from a three-dimensional (3D) culture possess higher therapeutic efficacy than conventional hMSCs from a monolayer (2D) culture. To date, various 3D culture methods have been developed to form hMSC spheroids but most of them used culture medium containing fetal bovine serum (FBS), which is not suitable for further clinical use. Here, we demonstrate that dissociated single MSCs seeded in induced pluripotent stem medium (MiPS) adhere loosely to the dish and spontaneously migrate to form spheroids during day 3 to day 6. Through component deletion screening and complementation experiments, the knockout serum replacement (KSR) was identified as necessary and sufficient for hMSC spheroid formation. Transcriptome analysis showed that the overall expression profiles were highly similar between 2D culture with FBS and KSR-derived spheroids. Interestingly, genes related to inflammatory response, immune response, and angiogenesis were upregulated in spheroids at day 6 and qPCR results further validated the increased expression level of related genes, including STC1, CCL7, HGF, IL24, and TGFB3. When spheroids were replated in normal FBS medium, cells formed a typical spindle-shaped morphology and FACS results showed that the recovered cells retained MSC-specific surface markers, such as CD73, CD90, and CD105. In summary, we developed a practical and convenient method to generate hMSC spheroids for clinical research and therapy.

scholarly journals Spheroid Formation and Evaluation of Hepatic Cells in a Three-Dimensional Culture Device

Cell Medicine ◽  
2015 ◽  
Vol 8 (1-2) ◽  
pp. 47-56 ◽  
Author(s):  
Yoshitaka Miyamoto ◽  
Masashi Ikeuchi ◽  
Hirofumi Noguchi ◽  
Tohru Yagi ◽  
Shuji Hayashi
Keyword(s):  
Three Dimensional ◽  
Spheroid Formation ◽  
Hepatic Cells ◽  
Three Dimensional Culture
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