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Cells
Special Issues
Primary and Continued Cell Cultures
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Primary and Continued Cell Cultures

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

Deadline for manuscript submissions: 8 July 2025 | Viewed by 14990

Special Issue Editor

Prof. Dr. Xuefeng Liu

E-Mail Website
Guest Editor
1. Center for Cell Reprograming, Departments of Pathology and Oncology, Georgetown University Medical Center, Washington, DC 20057, USA
2. Departments of Pathology and Urology, Comprehensive Cancer Center, The Ohio State University, Columbus, OH 43210, USA
Interests: cell reprogramming; telomerase and telomeres; patient-derived models; cell therapies; living biobanks; viruses; cancer
Special Issues, Collections and Topics in MDPI journals

Special Issue Information

Dear colleagues,

Traditional cancer cell lines and transformed cell lines are widely used in medical research. These cells usually have abnormalities resulting from the original tumor itself, but may also develop abnormalities due to genetic manipulation, or due to genetic and epigenetic changes during long-term passage. Primary cultures may maintain the lineage functions of their original tissue types, yet their life span and population doubling time are limited due to the nature of cellular senescence. Primary cultures usually have low yields and high variability compared to original tissue specimens, limiting their applications in research. Conditional cell reprogramming (CR) makes it possible to rapidly expand and continually culture primary cells from surgical specimens, core or needle biopsies, and other minimally invasive or noninvasive specimens—for example, nasal cavity brushing from a variety of species, including human, mouse, rat, horse, and ferret. These conditionally reprogrammed cells (CRCs) preserve their lineage functions and provide biologically relevant and physiological conditions suitable for studies of toxicity, virology, and tissue repair and the discovery of antiviral or anticancer drugs. This Special Issue will focus on primary cell cultures (protocols, reviews, and original articles) using conditionally reprogrammed cells, organoids, and other recent approaches.

Prof. Dr. Xuefeng Liu
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 100 words) can be sent to the Editorial Office for announcement on this website.

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.

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

  • primary normal cells
  • primary tumor cells
  • patient-derived cells
  • CRC (conditionally reprogrammed cells)
  • organoids
  • circulating tumor cells
  • human diseases
  • toxicity
  • cancer models
  • drug discovery

Published Papers (4 papers)

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Research

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17 pages, 3985 KiB  
Article
StemRegenin-1 Attenuates Endothelial Progenitor Cell Senescence by Regulating the AhR Pathway-Mediated CYP1A1 and ROS Generation
by Hye Ji Lim, Woong Bi Jang, Vinoth Kumar Rethineswaran, Jaewoo Choi, Eun Ji Lee, Sangmi Park, Yeoreum Jeong, Jong Seong Ha, Jisoo Yun, Young Jin Choi, Young Joon Hong and Sang-Mo Kwon
Cells 2023, 12(15), 2005; https://doi.org/10.3390/cells12152005 - 5 Aug 2023
Cited by 3 | Viewed by 1561
Abstract
Endothelial progenitor cell (EPC)-based stem cell therapy is a promising therapeutic strategy for vascular diseases. However, continuous in vitro expansion for clinical studies induces the loss of EPC functionality due to aging. In this study, we investigated the effects of StemRegenin-1 (SR-1), an [...] Read more.
Endothelial progenitor cell (EPC)-based stem cell therapy is a promising therapeutic strategy for vascular diseases. However, continuous in vitro expansion for clinical studies induces the loss of EPC functionality due to aging. In this study, we investigated the effects of StemRegenin-1 (SR-1), an antagonist of aryl hydrocarbon receptor (AhR), on replicative senescence in EPCs. We found that SR-1 maintained the expression of EPC surface markers, including stem cell markers, such as CD34, c-Kit, and CXCR4. Moreover, SR-1 long-term-treated EPCs preserved their characteristics. Subsequently, we demonstrated that SR-1 showed that aging phenotypes were reduced through senescence-associated phenotypes, such as β-galactosidase activity, SMP30, p21, p53, and senescence-associated secretory phenotype (SASP). SR-1 treatment also increased the proliferation, migration, and tube-forming capacity of senescent EPCs. SR-1 inhibited the AhR-mediated cytochrome P450 (CYP)1A1 expression, reactive-oxygen species (ROS) production, and DNA damage under oxidative stress conditions in EPCs. Furthermore, as a result of CYP1A1-induced ROS inhibition, it was found that accumulated intracellular ROS were decreased in senescent EPCs. Finally, an in vivo Matrigel plug assay demonstrated drastically enhanced blood vessel formation via SR-1-treated EPCs. In summary, our results suggest that SR-1 contributes to the protection of EPCs against cellular senescence. Full article
(This article belongs to the Special Issue Primary and Continued Cell Cultures)
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Review

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25 pages, 5252 KiB  
Review
Unlocking Translational Potential: Conditionally Reprogrammed Cells in Advancing Breast Cancer Research
by Danyal Daneshdoust, Mingjue Luo, Zaibo Li, Xiaokui Mo, Sahar Alothman, Bhaskar Kallakury, Richard Schlegel, Junran Zhang, Deliang Guo, Priscilla A. Furth, Xuefeng Liu and Jenny Li
Cells 2023, 12(19), 2388; https://doi.org/10.3390/cells12192388 - 30 Sep 2023
Cited by 1 | Viewed by 1555
Abstract
Preclinical in vitro models play an important role in studying cancer cell biology and facilitating translational research, especially in the identification of drug targets and drug discovery studies. This is particularly relevant in breast cancer, where the global burden of disease is quite [...] Read more.
Preclinical in vitro models play an important role in studying cancer cell biology and facilitating translational research, especially in the identification of drug targets and drug discovery studies. This is particularly relevant in breast cancer, where the global burden of disease is quite high based on prevalence and a relatively high rate of lethality. Predictive tools to select patients who will be responsive to invasive or morbid therapies (radiotherapy, chemotherapy, immunotherapy, and/or surgery) are relatively lacking. To be clinically relevant, a model must accurately replicate the biology and cellular heterogeneity of the primary tumor. Addressing these requirements and overcoming the limitations of most existing cancer cell lines, which are typically derived from a single clone, we have recently developed conditional reprogramming (CR) technology. The CR technology refers to a co-culture system of primary human normal or tumor cells with irradiated murine fibroblasts in the presence of a Rho-associated kinase inhibitor to allow the primary cells to acquire stem cell properties and the ability to proliferate indefinitely in vitro without any exogenous gene or viral transfection. This innovative approach fulfills many of these needs and offers an alternative that surpasses the deficiencies associated with traditional cancer cell lines. These CR cells (CRCs) can be reprogrammed to maintain a highly proliferative state and reproduce the genomic and histological characteristics of the parental tissue. Therefore, CR technology may be a clinically relevant model to test and predict drug sensitivity, conduct gene profile analysis and xenograft research, and undertake personalized medicine. This review discusses studies that have applied CR technology to conduct breast cancer research. Full article
(This article belongs to the Special Issue Primary and Continued Cell Cultures)
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23 pages, 5283 KiB  
Review
A Beginner’s Guide to Cell Culture: Practical Advice for Preventing Needless Problems
by Sabine Weiskirchen, Sarah K. Schröder, Eva Miriam Buhl and Ralf Weiskirchen
Cells 2023, 12(5), 682; https://doi.org/10.3390/cells12050682 - 21 Feb 2023
Cited by 8 | Viewed by 10043
Abstract
The cultivation of cells in a favorable artificial environment has become a versatile tool in cellular and molecular biology. Cultured primary cells and continuous cell lines are indispensable in investigations of basic, biomedical, and translation research. However, despite their important role, cell lines [...] Read more.
The cultivation of cells in a favorable artificial environment has become a versatile tool in cellular and molecular biology. Cultured primary cells and continuous cell lines are indispensable in investigations of basic, biomedical, and translation research. However, despite their important role, cell lines are frequently misidentified or contaminated by other cells, bacteria, fungi, yeast, viruses, or chemicals. In addition, handling and manipulating of cells is associated with specific biological and chemical hazards requiring special safeguards such as biosafety cabinets, enclosed containers, and other specialized protective equipment to minimize the risk of exposure to hazardous materials and to guarantee aseptic work conditions. This review provides a brief introduction about the most common problems encountered in cell culture laboratories and some guidelines on preventing or tackling respective problems. Full article
(This article belongs to the Special Issue Primary and Continued Cell Cultures)
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Other

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14 pages, 1703 KiB  
Commentary
Nasopharyngeal Carcinoma Cell Lines: Reliable Alternatives to Primary Nasopharyngeal Cells?
by Anna Makowska and Ralf Weiskirchen
Cells 2024, 13(7), 559; https://doi.org/10.3390/cells13070559 - 22 Mar 2024
Cited by 1 | Viewed by 972
Abstract
Nasopharyngeal carcinoma (NPC) is a type of cancer that originates from the mucosal lining of the nasopharynx and can invade and spread. Although contemporary chemoradiotherapy effectively manages the disease locally, there are still challenges with locoregional recurrence and distant failure. Therefore, it is [...] Read more.
Nasopharyngeal carcinoma (NPC) is a type of cancer that originates from the mucosal lining of the nasopharynx and can invade and spread. Although contemporary chemoradiotherapy effectively manages the disease locally, there are still challenges with locoregional recurrence and distant failure. Therefore, it is crucial to have a deeper understanding of the molecular basis of NPC cell movement in order to develop a more effective treatment and to improve patient survival rates. Cancer cell line models are invaluable in studying health and disease and it is not surprising that they play a critical role in NPC research. Consequently, scientists have established around 80 immortalized human NPC lines that are commonly used as in vitro models. However, over the years, it has been observed that many cell lines are misidentified or contaminated by other cells. This cross-contamination leads to the creation of false cell lines that no longer match the original donor. In this commentary, we discuss the impact of misidentified NPC cell lines on the scientific literature. We found 1159 articles from 2000 to 2023 that used NPC cell lines contaminated with HeLa cells. Alarmingly, the number of publications and citations using these contaminated cell lines continued to increase, even after information about the contamination was officially published. These articles were most commonly published in the fields of oncology, pharmacology, and experimental medicine research. These findings highlight the importance of science policy and support the need for journals to require authentication testing before publication. Full article
(This article belongs to the Special Issue Primary and Continued Cell Cultures)
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