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Cells
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Importance of Primary Cell Culture in Biomedical Research
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Importance of Primary Cell Culture in Biomedical Research

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

Deadline for manuscript submissions: closed (15 August 2023) | Viewed by 16631

Special Issue Editor

Prof. Dr. Yunsong Liu

E-Mail Website
Guest Editor
School of Stomatology, Peking University, Beijing, China
Interests: stem cell differentiation; tissue engineering; bone regeneration; translational research

Special Issue Information

Dear Colleagues,

Primary cell culture plays an important role in biomedical research. Changes in culturing conditions can regulate the biological behavior and performance of the cells, which can induce the cells into a more suitable state for use. Common methods include adding many kinds of regulating factors to the culture medium, such as cytokines, drugs, and biological cell products (e.g., extracellular vesicle). In this regard, modifying cells by changing the culture conditions is a promising therapeutic approach to improve the therapeutic effect of stem cells. This Special Issue aims at providing novel insights into the importance of primary cell culture in the context of translational research and biomedical application. We welcome original research and review articles in which the topics may include (but are not limited to) the following:

  • Methods in in vitro cell culture that can improve cell performance and their related mechanisms;
  • Therapeutic strategies aiming to enhance cell function by changing the cell culturing condition in the context of translational medicine, such as cytotherapy, tissue engineering, etc.;
  • Reviews that discuss recent advances and future perspectives in understanding the importance and role of primary cell culture in biomedical research.

We look forward to receiving your contributions.

Prof. Dr. Yunsong 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

  • stem cells
  • cell culture
  • regulating factors
  • cell products
  • biomedical application
  • translational research

Published Papers (6 papers)

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Research

Jump to: Review

14 pages, 6601 KiB  
Article
An Example of Neuro-Glial Commitment and Differentiation of Muse Stem Cells Obtained from Patients with IQSEC2-Related Neural Disorder: A Possible New Cell-Based Disease Model
by Sura Hilal Ahmed Al Sammarraie, Domenico Aprile, Ilaria Meloni, Nicola Alessio, Francesca Mari, Marianna Manata, Caterina Lo Rizzo, Giovanni Di Bernardo, Gianfranco Peluso, Alessandra Renieri and Umberto Galderisi
Cells 2023, 12(7), 977; https://doi.org/10.3390/cells12070977 - 23 Mar 2023
Cited by 1 | Viewed by 1332
Abstract
Although adult stem cells may be useful for studying tissue-specific diseases, they cannot be used as a general model for investigating human illnesses given their limited differentiation potential. Multilineage-differentiating stress-enduring (Muse) stem cells, a SSEA3(+) cell population isolated from mesenchymal stromal cells, fat, [...] Read more.
Although adult stem cells may be useful for studying tissue-specific diseases, they cannot be used as a general model for investigating human illnesses given their limited differentiation potential. Multilineage-differentiating stress-enduring (Muse) stem cells, a SSEA3(+) cell population isolated from mesenchymal stromal cells, fat, and skin fibroblasts, may be able to overcome that restriction. The Muse cells present in fibroblast cultures obtained from biopsies of patients’ skin may be differentiated into cells of interest for analyzing diseases. We isolated Muse stem cells from patients with an intellectual disability (ID) and mutations in the IQSEC2 gene (i.e., BRAG1 gene) and induced in vitro neuroglial differentiation to study cell commitment and the differentiation of neural lineages. The neuroglial differentiation of Muse cells revealed that IQSEC2 mutations may alter the self-renewal and lineage specification of stem cells. We observed a decrease in the percentage of SOX2 (+) neural stem cells and neural progenitors (i.e., SOX2+ and NESTIN+) in cultures obtained from Muse cells with the mutated IQSEC2 gene. The alteration in the number of stem cells and progenitors produced a bias toward the astrocytes’ differentiation. Our research demonstrates that Muse stem cells may represent a new cell-based disease model. Full article
(This article belongs to the Special Issue Importance of Primary Cell Culture in Biomedical Research)
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12 pages, 2956 KiB  
Article
Topical Application of Butyl Flufenamate Ointment Promotes Cranial Defect Healing in Mice by Inducing BMP2 Secretion in Skin Mesenchymal Stem Cells
by Fan Yang, Xuenan Liu, Donghao Wei, Yuan Zhu, Feilong Wang, Xuejiao Liu, Fanyu Yan, Xiao Zhang and Yunsong Liu
Cells 2022, 11(22), 3620; https://doi.org/10.3390/cells11223620 - 15 Nov 2022
Cited by 2 | Viewed by 1368
Abstract
Bone defects and fractures heal slowly compared with injuries to other tissues, creating a heavy burden for patients, their families, and society. Alongside conventional treatment methods for fractures and bone defects, adjuvant therapies play an important but underappreciated role. In a previous study, [...] Read more.
Bone defects and fractures heal slowly compared with injuries to other tissues, creating a heavy burden for patients, their families, and society. Alongside conventional treatment methods for fractures and bone defects, adjuvant therapies play an important but underappreciated role. In a previous study, we found that systemic administration of flufenamic acid promoted osteogenesis in vivo, but its side effects limited the application of our findings. In the present study, we assess the effects of external butyl flufenamate ointment on the healing of cranial defects in mice. We found that application of butyl flufenamate ointment on the surface of the skin accelerated the healing of cranial defects in mice by promoting BMP2 secretion from mouse-skin mesenchymal stem-cells. These findings indicate that butyl flufenamate ointment has potential therapeutic value for treating superficial fractures or bone defects while avoiding the toxicity and side effects of systemic medication, representing a safe and convenient adjuvant therapy to promote healing of superficial bone defects and fractures. Full article
(This article belongs to the Special Issue Importance of Primary Cell Culture in Biomedical Research)
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15 pages, 3227 KiB  
Article
Midpalatal Suture: Single-Cell RNA-Seq Reveals Intramembrane Ossification and Piezo2 Chondrogenic Mesenchymal Cell Involvement
by Lu Gao, Tiansong Xu, Liqi Zhang, Yuchen Li, Tianxing Yan, Guoxia Yu and Feng Chen
Cells 2022, 11(22), 3585; https://doi.org/10.3390/cells11223585 - 12 Nov 2022
Cited by 3 | Viewed by 2168
Abstract
The midpalatal suture is mainly responsible for the growth and development of the maxillary and resistance to rapid maxillary expansion (RME). It is essential for clinical researchers to explore the intramembrane ossification and to elucidate the underlying mechanism of the maturation and ossification [...] Read more.
The midpalatal suture is mainly responsible for the growth and development of the maxillary and resistance to rapid maxillary expansion (RME). It is essential for clinical researchers to explore the intramembrane ossification and to elucidate the underlying mechanism of the maturation and ossification process of the midpalatal suture to help identify the optimum time and force of RME. However, mechanistic studies associated with the midpalatal suture are rare. The aim of this present study is to create an intramembrane osteogenesis model for the midpalatal suture region of mice. Interestingly, we discovered a type of chondrogenic mesenchymal cell expressing Piezo2, which might be related to the detection of mechanical and external stimuli. This result provides a potential molecular and cellular mechanism that explains why the midpalatal suture is not closed until adulthood. We depict a landscape of mesenchymal cells that might play an important role in the intramembrane osteogenesis of the midpalatal suture and provide new perspectives on midpalate suture maturation and ossification, which might lead to further possibilities for clinical operations. Full article
(This article belongs to the Special Issue Importance of Primary Cell Culture in Biomedical Research)
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16 pages, 4940 KiB  
Article
In Vitro and In Vivo Studies of Hydrogenated Titanium Dioxide Nanotubes with Superhydrophilic Surfaces during Early Osseointegration
by Caiyun Wang, Shang Gao, Ran Lu, Xin Wang and Su Chen
Cells 2022, 11(21), 3417; https://doi.org/10.3390/cells11213417 - 28 Oct 2022
Cited by 5 | Viewed by 1569
Abstract
Titanium-based implants are often utilized in oral implantology and craniofacial reconstructions. However, the biological inertness of machined titanium commonly results in unsatisfactory osseointegration. To improve the osseointegration properties, we modified the titanium implants with nanotubular/superhydrophilic surfaces through anodic oxidation and thermal hydrogenation and [...] Read more.
Titanium-based implants are often utilized in oral implantology and craniofacial reconstructions. However, the biological inertness of machined titanium commonly results in unsatisfactory osseointegration. To improve the osseointegration properties, we modified the titanium implants with nanotubular/superhydrophilic surfaces through anodic oxidation and thermal hydrogenation and evaluated the effects of the machined surfaces (M), nanotubular surfaces (Nano), and hydrogenated nanotubes (H-Nano) on osteogenesis and osseointegration in vitro and in vivo. After incubation of mouse bone marrow mesenchymal stem cells on the samples, we observed improved cell adhesion, alkaline phosphatase activity, osteogenesis-related gene expression, and extracellular matrix mineralization in the H-Nano group compared to the other groups. Subsequent in vivo studies indicated that H-Nano implants promoted rapid new bone regeneration and osseointegration at 4 weeks, which may be attributed to the active osteoblasts adhering to the nanotubular/superhydrophilic surfaces. Additionally, the Nano group displayed enhanced osteogenesis in vitro and in vivo at later stages, especially at 8 weeks. Therefore, we report that hydrogenated superhydrophilic nanotubes can significantly accelerate osteogenesis and osseointegration at an early stage, revealing the considerable potential of this implant modification for clinical applications. Full article
(This article belongs to the Special Issue Importance of Primary Cell Culture in Biomedical Research)
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13 pages, 2258 KiB  
Article
Isolation of Human Osteoblast Cells Capable for Mineralization and Synthetizing Bone-Related Proteins In Vitro from Adult Bone
by Daria Kostina, Arseniy Lobov, Polina Klausen, Vitaly Karelkin, Rashid Tikhilov, Svetlana Bozhkova, Andrey Sereda, Nadezhda Ryumina, Natella Enukashvily and Anna Malashicheva
Cells 2022, 11(21), 3356; https://doi.org/10.3390/cells11213356 - 24 Oct 2022
Cited by 3 | Viewed by 4527
Abstract
The culture of osteoblasts (OB) of human origin is a useful experimental model in studying bone biology, osteogenic differentiation, functions of bone proteins, oncological processes in bone tissue, testing drugs against bone desires, and many other fields. The purpose of the present study [...] Read more.
The culture of osteoblasts (OB) of human origin is a useful experimental model in studying bone biology, osteogenic differentiation, functions of bone proteins, oncological processes in bone tissue, testing drugs against bone desires, and many other fields. The purpose of the present study is to share a workflow that has established the conditions to efficiently isolate and grow OB cells obtained from surgically removed bones from human donors. The protocol described here also shows how to determine cell phenotype. Here we provide characteristics of cells isolated by this protocol that might help researchers to decide if such OB are suitable for the purposes of their study. Osteoblasts isolated from collagenase-treated explants of adult bones are able to proliferate and keep their phenotype in culture. OB cells have high synthetic properties. They express osteomarkers, such as RUNX2, osteocalcin, BMP2, and osteopontin both in control conditions and in an osteogenic medium that could be estimated by qPCR and immunocytochemical staining and by Western blotting. Induction of osteogenic differentiation does not dramatically influence the synthetic properties of OB cells, while the cells gain the ability to extracellular mineralization only in an osteogenic medium. Full article
(This article belongs to the Special Issue Importance of Primary Cell Culture in Biomedical Research)
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Review

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21 pages, 855 KiB  
Review
Rejuvenation of Mesenchymal Stem Cells to Ameliorate Skeletal Aging
by Mingjia Cheng, Weihao Yuan, Alireza Moshaverinia and Bo Yu
Cells 2023, 12(7), 998; https://doi.org/10.3390/cells12070998 - 24 Mar 2023
Cited by 4 | Viewed by 3749
Abstract
Advanced age is a shared risk factor for many chronic and debilitating skeletal diseases including osteoporosis and periodontitis. Mesenchymal stem cells develop various aging phenotypes including the onset of senescence, intrinsic loss of regenerative potential and exacerbation of inflammatory microenvironment via secretory factors. [...] Read more.
Advanced age is a shared risk factor for many chronic and debilitating skeletal diseases including osteoporosis and periodontitis. Mesenchymal stem cells develop various aging phenotypes including the onset of senescence, intrinsic loss of regenerative potential and exacerbation of inflammatory microenvironment via secretory factors. This review elaborates on the emerging concepts on the molecular and epigenetic mechanisms of MSC senescence, such as the accumulation of oxidative stress, DNA damage and mitochondrial dysfunction. Senescent MSCs aggravate local inflammation, disrupt bone remodeling and bone-fat balance, thereby contributing to the progression of age-related bone diseases. Various rejuvenation strategies to target senescent MSCs could present a promising paradigm to restore skeletal aging. Full article
(This article belongs to the Special Issue Importance of Primary Cell Culture in Biomedical Research)
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