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Biology
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In Vitro 2.0—Improving the Cell Culture Environment for Biology
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In Vitro 2.0—Improving the Cell Culture Environment for Biology

A special issue of Biology (ISSN 2079-7737). This special issue belongs to the section "Cell Biology".

Deadline for manuscript submissions: 31 March 2026 | Viewed by 1781

Special Issue Editor

Dr. Marcus Krüger

E-Mail Website
Guest Editor
Environmental Cell Biology Group, Department of Microgravity and Translational Regenerative Medicine, Otto von Guericke University, 39106 Magdeburg, Germany
Interests: cell biology; cancer biology; environmental influences; microgravity; cellular communication; photodynamic therapy; cancer treatment; antimicrobial resistance; tumor microbiome
Special Issues, Collections and Topics in MDPI journals

Special Issue Information

Dear Colleagues,

In vitro studies are still the basis for biological research. Although scientists have considered static culture in plastic bottles as the gold standard of in vitro studies for decades, a better understanding of molecular biological processes and the presence of new technologies (microfluidics, tissue engineering, advanced bioreactors, bioprinting, etc.) are leading to an optimization of in vitro cell cultures that allow for more reliable data required for the prediction of biological effects in organisms. The ability to grow and change in a defined manner in response to various stimuli is one of the most remarkable properties of cells. It plays an important role in morphogenesis during development and in homeostasis and pathogenesis in adult tissues, which often adapt to changes in their mechanochemical environment due to aging, disease, or injury.

We have now reached a point where we can offer customized cell culture conditions for specific research purposes. With this Special Issue, we would like to review the status quo of cell culture science, the latest findings, and improved approaches from which we can all benefit. Review articles, original research papers, short communications, and comments are all welcome for submission. The focus of these submissions should be on the practical application of techniques and is not limited to human research only.

We look forward to your submissions.

Dr. Marcus Krüger
Guest Editor

Manuscript Submission Information

Manuscripts should be submitted online at www.mdpi.com by registering and logging in to this website. Once you are registered, click here to go to the submission form. Manuscripts can be submitted until the deadline. All submissions that pass pre-check are peer-reviewed. Accepted papers will be published continuously in the journal (as soon as accepted) and will be listed together on the special issue website. Research articles, review articles as well as short communications are invited. For planned papers, a title and short abstract (about 250 words) can be sent to the Editorial Office for assessment.

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. Biology is an international peer-reviewed open access monthly journal published by MDPI.

Please visit the Instructions for Authors page before submitting a manuscript. The Article Processing Charge (APC) for publication in this open access journal is 2700 CHF (Swiss Francs). Submitted papers should be well formatted and use good English. Authors may use MDPI's English editing service prior to publication or during author revisions.

Keywords

  • in vitro technologies
  • microfluids
  • bioreactors
  • organoids
  • tissue engineering
  • lab-on-a-chip
  • life-support systems
  • symbioses

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Published Papers (2 papers)

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Research

21 pages, 3276 KB  
Article
Construction and Performance Evaluation of an Astaxanthin–Chitosan/Chitooligosaccharide Hydrogel System for Ex Vivo Culture of Murine Spermatogonial Stem Cells
by Jiang Wu, Siqi Liu, Xiaowen Zeng, Yang Li, Yinlin Yao, Jing Wang, Guangdong Hu and Kai Kang
Biology 2025, 14(12), 1664; https://doi.org/10.3390/biology14121664 - 24 Nov 2025
Viewed by 328
Abstract
Spermatogonial stem cells (SSCs) are the only adult male germline stem cells capable of lifelong self-renewal and differentiation into spermatozoa. Scalable ex vivo survival is essential for endangered species germplasm banking, genetic resource conservation, and male infertility therapy. Here, chitosan (CO) or chitosan [...] Read more.
Spermatogonial stem cells (SSCs) are the only adult male germline stem cells capable of lifelong self-renewal and differentiation into spermatozoa. Scalable ex vivo survival is essential for endangered species germplasm banking, genetic resource conservation, and male infertility therapy. Here, chitosan (CO) or chitosan oligosaccharide (COS) was cross-linked into injectable, biodegradable 3D hydrogels loaded with the natural astaxanthin (AST). CCK-8 optimization identified 0.3% CO + 0.2% AST (CHAG) and 0.2% COS + 0.2% AST (COAG) as superior formulations. After 7 or 14 d of 3D culture, CHAG yielded significantly more colonies than controls (p < 0.01), with elevated EdU incorporation, alkaline phosphatase activity, and positive OCT4 and PLZF staining, confirming preserved stemness. Caspase-3 expression was markedly reduced, indicating the AST-mediated suppression of oxidative apoptosis. RNA-seq showed distinct transcriptome pathways (p < 0.01): CHAG up-regulated adhesion and ECM–receptor and cell cycle pathways, whereas COAG enriched immune-modulatory and signaling modules, enabling context-specific use. AST-loaded CO/COS hydrogels are inexpensive, cytocompatible, and scalable, doubling as a biomimetic niche that accelerates SSC proliferation while delaying senescence. The platform provides a robust, controllable 3D system for SSC expansion and establishes a pre-clinical basis for translating CO/COS/AST composites to reproductive stem cell biotechnology. Full article
(This article belongs to the Special Issue In Vitro 2.0—Improving the Cell Culture Environment for Biology)
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15 pages, 2260 KB  
Article
Efficient Serum-Free Rabies Virus Propagation Using BSR and Vero Cell Lines: A Comparative Evaluation of BioNOC II® Macrocarriers in the BelloStage™-3000 Bioreactor Versus Conventional Microcarriers
by Zhanat Amanova, Zhanna Sametova, Sholpan Turyskeldy, Alina Kurmasheva, Ruslan Abitayev, Abdurakhman Ussembay, Zhanat Kondibaeva, Dariya Toktyrova, Dana Mazbayeva, Sergazy Nurabayev, Aslan Kerimbayev and Yerbol Bulatov
Biology 2025, 14(10), 1455; https://doi.org/10.3390/biology14101455 - 21 Oct 2025
Viewed by 1129
Abstract
The rabies virus remains a significant public health threat, particularly in regions with limited access to vaccination. This study shows that the BelloStage™-3000 bioreactor, operating on the “Tide Motion” principle, in combination with BioNOC® II macrocarriers, ensures highly efficient rabies virus cultivation [...] Read more.
The rabies virus remains a significant public health threat, particularly in regions with limited access to vaccination. This study shows that the BelloStage™-3000 bioreactor, operating on the “Tide Motion” principle, in combination with BioNOC® II macrocarriers, ensures highly efficient rabies virus cultivation in BSR and Vero cells grown in serum-free OptiPRO™ SFM medium. This system supports effective cell attachment, formation of a dense and metabolically active cell layer, and reduces microbial contamination risks associated with serum-containing media. For comparison, rabies virus cultivation was also performed on Cytodex 1 and Cytodex 3 microcarriers in spinner flasks. The use of the BelloStage™-3000 bioreactor system with BelloCell™ 500A disposable vials and BioNOC II® macrocarriers resulted in significantly higher virus titers compared to traditional Cytodex 1 and Cytodex 3 microcarrier culture systems. Thus, in the BSR cell culture, the maximum virus titer reached 5.6 × 108 FFU/mL by day 4 of cultivation, which exceeded the titers obtained on Cytodex 1 and Cytodex 3 microcarriers by about 19.3-fold and 15.3-fold, respectively. A similar trend was observed for the Vero cell line: the peak titer was 2.0 × 108 FFU/mL by day 5 of culturing, which was higher than the values obtained on Cytodex 1 and Cytodex 3 by about 14.0-fold and 9.6-fold, respectively. These findings demonstrate that the integrated use of BioNOC® II macrocarriers, the BelloStage™-3000 bioreactor, and a serum-free medium provides a scalable, reproducible, and biosafe platform for rabies virus production, offering substantial advantages over traditional microcarrier-based systems. Full article
(This article belongs to the Special Issue In Vitro 2.0—Improving the Cell Culture Environment for Biology)
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