3D Stem Cell Culture—Series 2

A special issue of Cells (ISSN 2073-4409). This special issue belongs to the section "Stem Cells".

Deadline for manuscript submissions: closed (31 January 2024) | Viewed by 2744

Special Issue Editor


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Guest Editor
Department of Biology, University of Mary Hardin-Baylor, 900 College Street, Box 8432, Belton, TX 76513, USA
Interests: mesenchymal stem cell (MSC) biology; 3D-cell culture; microbiology; immunology; regenerative medicine
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Special Issue Information

Dear Colleagues,

Stem cells have received extensive interest recently because of their unique characteristics and applications in basic and preclinical research. The extent of stem cell use in clinical trials has also increased tremendously owing to their promising therapeutic effects in animal models and in patients. Numerous different stem cells have been isolated from various adult and fetal tissues, and these cells are able to self-renew and differentiate into various end-stage cell types. In order to generate enough stem cells for research and therapies, cells must often be culture-expanded. These cultures often employ traditional 2D techniques that do not necessarily mimic the stem cell environment in vivo well. Therefore, there is great interest in developing 3D culture techniques and characterizing stem cells grown under these conditions to understand basic stem cell biology and also develop effective cells for therapies. These 3D cultures might hold the answers for optimal stem cell preparations for clinical applications.

We invite investigators to contribute reviews and original papers that describe recent findings in the field of 3D stem cell cultures. We especially encourage the submission of research regarding various 3D culture techniques and regenerative medicine approaches using in vitro and in vivo models.

You may choose our Joint Special Issue in Organoids.

Dr. Joni H. Ylostalo
Guest Editor

Manuscript Submission Information

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Submitted manuscripts should not have been published previously, nor be under consideration for publication elsewhere (except conference proceedings papers). All manuscripts are thoroughly refereed through a single-blind peer-review process. A guide for authors and other relevant information for submission of manuscripts is available on the Instructions for Authors page. Cells is an international peer-reviewed open access semimonthly journal published by MDPI.

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Keywords

  • stem cell
  • 3D
  • culture condition
  • expansion
  • growth
  • characteristic
  • niche
  • regenerative medicine
  • animal model

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Published Papers (3 papers)

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20 pages, 2904 KiB  
Article
Unlocking the Potential of Human-Induced Pluripotent Stem Cells: Cellular Responses and Secretome Profiles in Peptide Hydrogel 3D Culture
by Muyun Cui, Wei Wu, Quan Li, Guangyan Qi, Xuming Liu, Jianfa Bai, Mingshun Chen, Ping Li and Xiuzhi (Susan) Sun
Cells 2024, 13(2), 143; https://doi.org/10.3390/cells13020143 - 12 Jan 2024
Cited by 1 | Viewed by 1057
Abstract
Human-induced pluripotent stem cells (hiPSCs) have shown great potential for human health, but their growth and properties have been significantly limited by the traditional monolayer (2D) cell culture method for more than 15 years. Three-dimensional (3D) culture technology has demonstrated tremendous advantages over [...] Read more.
Human-induced pluripotent stem cells (hiPSCs) have shown great potential for human health, but their growth and properties have been significantly limited by the traditional monolayer (2D) cell culture method for more than 15 years. Three-dimensional (3D) culture technology has demonstrated tremendous advantages over 2D. In particular, the 3D PGmatrix hiPSC derived from a peptide hydrogel offers a breakthrough pathway for the maintenance and expansion of physiologically relevant hiPSC 3D colonies (spheroids). In this study, the impact of 3D culture conditions in PGmatrix hiPSC on cell performance, integrity, and secretome profiles was determined across two commonly used hiPSC cell lines derived from fibroblast cells (hiPSC-F) and peripheral blood mononuclear cells (hiPSC-P) in the two most popular hiPSC culture media (mTeSR1 and essential eight (E8)). The 3D culture conditions varied in hydrogel strength, 3D embedded matrix, and 3D suspension matrix. The results showed that hiPSCs cultured in 3D PGmatrix hiPSC demonstrated the ability to maintain a consistently high cell viability that was above 95% across all the 3D conditions with cell expansion rates of 10–20-fold, depending on the 3D conditions and cell lines. The RT-qPCR analysis suggested that pluripotent gene markers are stable and not significantly affected by the cell lines or 3D PGmatrix conditions tested in this study. Mass spectrometry-based analysis of secretome from hiPSCs cultured in 3D PGmatrix hiPSC revealed a significantly higher quantity of unique proteins, including extracellular vesicle (EV)-related proteins and growth factors, compared to those in the 2D culture. Moreover, this is the first evidence to identify that hiPSCs in a medium with a rich supplement (i.e., mTeSR1) released more growth-regulating factors, while in a medium with fewer supplements (i.e., E8) hiPSCs secreted more survival growth factors and extracellular proteins. These findings offer insights into how these differences may impact hiPSC behavior, and they deepen our understanding of how hiPSCs respond to 3D culture conditions, aiding the optimization of hiPSC properties in translational biomedical research toward clinical applications. Full article
(This article belongs to the Special Issue 3D Stem Cell Culture—Series 2)
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17 pages, 8375 KiB  
Article
Resveratrol’s Impact on the Chondrogenic Reagents’ Effects in Cell Sheet Cultures of Wharton’s Jelly-Derived MSCs
by Anastasiia D. Kurenkova, Viktoria S. Presniakova, Zlata A. Mosina, Pavel D. Kibirskiy, Irina A. Romanova, Gilyana K. Tugaeva, Nastasia V. Kosheleva, Kirill S. Vinogradov, Sergei V. Kostjuk, Svetlana L. Kotova, Yury A. Rochev, Ekaterina V. Medvedeva and Peter S. Timashev
Cells 2023, 12(24), 2845; https://doi.org/10.3390/cells12242845 - 15 Dec 2023
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Abstract
Human Wharton’s jelly mesenchymal stem cells (hWJ-MSCs) are of great interest in tissue engineering. We obtained hWJ-MSCs from four patients, and then we stimulated their chondrogenic phenotype formation in vitro by adding resveratrol (during cell expansion) and a canonical Wnt pathway activator, LiCl, [...] Read more.
Human Wharton’s jelly mesenchymal stem cells (hWJ-MSCs) are of great interest in tissue engineering. We obtained hWJ-MSCs from four patients, and then we stimulated their chondrogenic phenotype formation in vitro by adding resveratrol (during cell expansion) and a canonical Wnt pathway activator, LiCl, as well as a Rho-associated protein kinase inhibitor, Y27632 (during differentiation). The effects of the added reagents on the formation of hWJ-MSC sheets destined to repair osteochondral injuries were investigated. Three-dimensional hWJ-MSC sheets grown on P(NIPAM-co-NtBA)-based matrices were characterized in vitro and in vivo. The combination of resveratrol and LiCl showed effects on hWJ-MSC sheets similar to those of the basal chondrogenic medium. Adding Y27632 decreased both the proportion of hypertrophied cells and the expression of the hyaline cartilage markers. In vitro, DMSO was observed to impede the effects of the chondrogenic factors. The mouse knee defect model experiment revealed that hWJ-MSC sheets grown with the addition of resveratrol and Y27632 were well integrated with the surrounding tissues; however, after 3 months, the restored tissue was identical to that of the naturally healed cartilage injury. Thus, the combination of chondrogenic supplements may not always have additive effects on the progress of cell culture and could be neutralized by the microenvironment after transplantation. Full article
(This article belongs to the Special Issue 3D Stem Cell Culture—Series 2)
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11 pages, 12740 KiB  
Brief Report
Generation of Highly Functional Hepatocyte-like Organoids from Human Adipose-Derived Mesenchymal Stem Cells Cultured with Endothelial Cells
by Shuhai Chen, Yu Saito, Yuhei Waki, Tetsuya Ikemoto, Hiroki Teraoku, Shinichiro Yamada, Yuji Morine and Mitsuo Shimada
Cells 2024, 13(6), 547; https://doi.org/10.3390/cells13060547 - 20 Mar 2024
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Abstract
Previously, we successfully established a highly functional, three-dimensional hepatocyte-like cell (3D-HLC) model from adipose-derived mesenchymal stem cells (ADSCs) via a three-step differentiation protocol. The aim of the present study was to investigate whether generating hepatocyte-like organoids (H-organoids) by adding endothelial cells further improved [...] Read more.
Previously, we successfully established a highly functional, three-dimensional hepatocyte-like cell (3D-HLC) model from adipose-derived mesenchymal stem cells (ADSCs) via a three-step differentiation protocol. The aim of the present study was to investigate whether generating hepatocyte-like organoids (H-organoids) by adding endothelial cells further improved the liver-like functionality of 3D-HLCs and to assess H-organoids’ immunogenicity properties. Genes representing liver maturation and function were detected by quantitative reverse transcription–PCR analysis. The expression of hepatic maturation proteins was measured using immunofluorescence staining. Cytochrome P (CYP)450 metabolism activity and ammonia metabolism tests were used to assess liver function. H-organoids were successfully established by adding human umbilical vein endothelial cells at the beginning of the definitive endoderm stage in our 3D differentiation protocol. The gene expression of alpha-1 antitrypsin, carbamoyl–phosphate synthase 1, and apolipoprotein E, which represent liver maturation state and function, was higher in H-organoids than non-organoid 3D-HLCs. H-organoids possessed higher CYP3A4 metabolism activity and comparable ammonia metabolism capacity than 3D-HLCs. Moreover, although H-organoids expressed human leukocyte antigen class I, they expressed little human leukocyte antigen class II, cluster of differentiation (CD)40, CD80, CD86, and programmed cell death ligand 1, suggesting their immunogenicity properties were not significantly upregulated during differentiation from ADSCs. In conclusion, we successfully established an H-organoid model with higher liver-like functionality than previously established 3D-HLCs and comparable immunogenicity to ADSCs. Full article
(This article belongs to the Special Issue 3D Stem Cell Culture—Series 2)
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