• Combined modality therapy
• Diabetes complications
• Immunomodulation
• Mesenchymal stem cells
• Type 1 diabetes

• Diabetes Mellitus, Type 1/therapy
• Diabetes Mellitus, Type 1/immunology
• Diabetes Mellitus, Type 1/complications
• Humans
• Mesenchymal Stem Cells/immunology
• Mesenchymal Stem Cells/cytology
• Mesenchymal Stem Cell Transplantation
• Animals
• Immunomodulation

• 82172230 Innovative Research Group Project of the National Natural Science Foundation of China
• 20190902007TC Jilin Province Science and Technology Development Plan Project
• 20210472. the Life Spring AKY Pharmmaceuticals Corporation

• Epithelial-to-mesenchymal transition
• Exosome modification
• Ferroptosis
• MSC-EXOs
• Mesenchymal stem cell

• Ferroptosis/physiology
• Ferroptosis/genetics
• Humans
• Exosomes/metabolism
• MicroRNAs/genetics
• MicroRNAs/metabolism
• MicroRNAs/administration & dosage
• Mesenchymal Stem Cells/metabolism
• Animals
• Neoplasms/therapy
• Neoplasms/genetics
• Neoplasms/pathology
• Tumor Microenvironment

• circRNA
• cord blood
• exosomes
• extracellular vesicles
• human-induced pluripotent stem cells
• miRNA
• nanoparticles

• Humans
• Induced Pluripotent Stem Cells/metabolism
• Induced Pluripotent Stem Cells/cytology
• Extracellular Vesicles/metabolism
• MicroRNAs/metabolism
• MicroRNAs/genetics
• RNA, Circular/metabolism
• RNA, Circular/genetics
• Mesenchymal Stem Cells/metabolism
• Mesenchymal Stem Cells/cytology
• Fetal Blood/cytology
• Cell Differentiation

• Mesenchymal stem cells
• Rejuvenation
• Senescence

• Humans
• Mesenchymal Stem Cells/metabolism
• Mesenchymal Stem Cells/cytology
• Cellular Senescence
• Cell Differentiation
• Animals
• Aging
• Cell Proliferation/genetics

• Autoimmune diseases
• Chronic inflammatory illnesses
• Clinical use of mesenchymal stem cell
• Mesenchymal stem cells
• Metastasis
• Regenerative medicine
• Therapeutic potential
• Transplantation controversies
• Tumor growth

• differentiation
• stem cells
• developmental biology
• cell biology

• Animals
• Mesenchymal Stem Cells/metabolism
• Mesenchymal Stem Cells/cytology
• Mice
• Cell Differentiation
• Cells, Cultured
• Transcriptome
• Bone Marrow Cells/cytology
• Bone Marrow Cells/metabolism
• Cell Culture Techniques/methods
• Gene Expression Profiling
• Mice, Inbred C57BL
• Cell Proliferation
• Cell Cycle

• RSP-312-19/20 Tour de Cure
• Australian Government Research Training Program
• Child Cancer Research Foundation
• Priority-driven Collaborative Cancer Research Scheme, and co-funded by Cancer Australia, Cure Cancer and the Leukaemia Foundation of Australia.

• Mesenchymal stem cell
• exosomes
• microRNA
• radiation injury

• Exosomes/metabolism
• Mesenchymal Stem Cells/radiation effects
• Mesenchymal Stem Cells/metabolism
• MicroRNAs/metabolism
• Humans
• Radiation Injuries/therapy
• Animals

• 3D culture
• Cellhesion® chitin nanoscaffolds
• Mesenchymal stem cells
• Stemness

• Differentiation
• Stem cells
• Stemness
• UC-MSCs
• Umbilical cord

• CTDSPL
• mesenchymal stem cells
• miR-18a-5p
• senescence

• MicroRNAs/genetics
• MicroRNAs/metabolism
• Cell Differentiation/genetics
• Cellular Senescence/genetics
• Up-Regulation
• Mesenchymal Stem Cells/metabolism

• CRISPR/Cas
• FVIII
• cell therapy
• gene therapy
• hemophilia A
• lentiviral (LV) vector
• placental-derived mesenchymal stromal cells

• Female
• Humans
• Pregnancy
• Hemophilia A/therapy
• Factor VIII
• Endothelial Cells/metabolism
• Placenta/metabolism
• Mesenchymal Stem Cells/metabolism

• R01 HL135853 NHLBI NIH HHS
• U01 HL148681 NHLBI NIH HHS
• Howard Hughes Medical Institute
• National Heart, Lung, and Blood Institute

Rapidly expanding clones (RECs) are one of the single-cell-derived mesenchymal stem cell clones sorted from human bone marrow mononuclear cells (BMMCs), which possess advantageous features. The RECs exhibit long-lasting proliferation potency that allows more than 10 repeated serial passages in vitro, considerably benefiting the manufacturing process of allogenic MSC-based therapeutic products. Although RECs aid the preparation of large-variation clone libraries for a greedy selection of better-quality clones, such a selection is only possible by establishing multiple-candidate cell banks for quality comparisons. Thus, there is a high demand for a novel method that can predict “low-risk and high-potency clones” early and in a feasible manner given the excessive cost and effort required to maintain such an establishment.

LNGFR and Thy-1 co-positive cells from BMMCs were single-cell-sorted into 96-well plates, and only fast-growing clones that reached confluency in 2 weeks were picked up and passaged as RECs. Fifteen RECs were prepared as passage 3 (P3) cryostock as the primary cell bank. From this cryostock, RECs were passaged until their proliferation limitation; their serial-passage limitation numbers were labeled as serial-passage potencies. At the P1 stage, phase-contrast microscopic images were obtained over 6–90 h to identify time-course changes of 24 morphological descriptors describing cell population information. Machine learning models were constructed using the morphological descriptors for predicting serial-passage potencies. The time window and field-of-view-number effects were evaluated to identify the most efficient image data usage condition for realizing high-performance serial-passage potency models.

Serial-passage test results indicated variations of 7–13-repeated serial-passage potencies within RECs. Such potency values were predicted quantitatively with high performance (RMSE < 1.0) from P1 morphological profiles using a LASSO model. The earliest and minimum effort predictions require 6–30 h with 40 FOVs and 6–90 h with 15 FOVs, respectively.

We successfully developed a noninvasive morphology-based machine learning model to enhance the efficiency of establishing cell banks with single-cell-derived RECs for quantitatively predicting the future serial-passage potencies of clones. Conventional methods that can make noninvasive and quantitative predictions without wasting precious cells in the early stage are lacking; the proposed method will provide a more efficient and robust cell bank establishment process for allogenic therapeutic product manufacturing.

The online version contains supplementary material available at 10.1186/s41232-022-00214-w.

• Cell bank establishment
• LNGFR
• Mesenchymal stem cells
• Morphological analysis
• Prediction model
• Rapidly expanding clone (REC)
• Serial-passage potency
• THY-1

• 3D tissue networks
• Cultured meat
• Porcine muscle satellite cells
• Reactive oxygen species
• l-ascorbic acid 2-phosphate

• Ministero della Salute

• cell engineering
• cell therapy
• drug delivery
• induced pluripotent stem cells
• mesenchymal stem cells
• preconditioning
• regenerative medicine
• scaffolds
• secretome

• CP19/00046 Instituto de Salud Carlos III
• PI20/00341 Instituto de Salud Carlos III
• IDEAS20051CAPI Asociación Española Contra el Cáncer
• PI-0272-2017 Regional Government of Andalusia
• PY20_00481 Regional Government of Andalusia
• FPU19/04703 Government of Spain
• +VIDA project Asociación Pablo Ugarte

Bone marrow aging is associated with multiple cellular dysfunctions, including perturbed haematopoiesis, the propensity to haematological transformation, and the maintenance of leukaemia. It has been shown that instructive signals from different leukemic cells are delivered to stromal cells to remodel the bone marrow into a supportive leukemic niche. In particular, cellular senescence, a physiological program with both beneficial and deleterious effects on the health of the organisms, may be responsible for the increased incidence of haematological malignancies in the elderly and for the survival of diverse leukemic cells. Here, we will review the connection between BM aging and cellular senescence and the role that these processes play in leukaemia progression. Specifically, we discuss the role of mesenchymal stem cells as a central component of the supportive niche. Due to the specificity of the genetic defects present in leukaemia, one would think that bone marrow alterations would also have particular changes, making it difficult to envisage a shared therapeutic use. We have tried to summarize the coincident features present in BM stromal cells during aging and senescence and in two different leukaemias, acute myeloid leukaemia, with high frequency in the elderly, and B-acute lymphoblastic leukaemia, mainly a childhood disease. We propose that mesenchymal stem cells are similarly affected in these different leukaemias, and that the changes that we observed in terms of cellular function, redox balance, genetics and epigenetics, soluble factor repertoire and stemness are equivalent to those occurring during BM aging and cellular senescence. These coincident features may be used to explore strategies useful to treat various haematological malignancies.

• angiogenesis
• cell therapy
• endothelial dysfunction
• stem cells
• vascularization

• Animals
• Endothelium, Vascular/metabolism
• Microvessels
• Oxidative Stress
• Regeneration
• Vascular Diseases/metabolism

• P30 ES030283 NIEHS NIH HHS
• R01 AG049821 NIA NIH HHS
• R01 AG053585 NIA NIH HHS
• Gheen's Foundation
• Canadian Network for Research and Innovation in Machining Technology, Natural Sciences and Engineering Research Council of Canada (CRSNG)
• Gouvernement du Canada | Canadian Institutes of Health Research (CIHR)
• HHS | NIH | National Institute on Aging (NIA)
• Ministère du Développement économique, de la Création d'emplois et du Commerce | Ontario Ministry of Research and Innovation (MRI)
• U.S. Department of Defense (DOD)

• 3D cell culture
• expansion
• hypoxia
• isolation
• mesenchymal stem/stromal cells

• German Research Foundation

• adipose-derived mesenchymal stromal cell
• bone-marrow derived mesenchymal stromal cell
• cartilage regeneration
• culture
• knee osteoarthritis
• mesenchymal stromal cell
• meta-analysis

• NRF-2019R1I1A1A01061296 National Research Foundation of Korea
• NRF-2021R1I1A1A01040732 National Research Foundation of Korea

• Nox1
• Rag2
• antioxidants
• diabetes
• mesenchymal stem cells
• oxidative stress

• Acetylcysteine/pharmacology
• Animals
• Antioxidants/metabolism
• Antioxidants/pharmacology
• Ascorbic Acid/pharmacology
• Diabetes Mellitus, Experimental/metabolism
• Diabetes Mellitus, Type 2/pathology
• Dietary Supplements
• Humans
• Male
• Mesenchymal Stem Cells/metabolism
• Mice
• Mice, Inbred C57BL
• Oxidative Stress
• Phosphates
• Quality of Life

Systemic lupus erythematosus (SLE) is a chronic autoimmune disease. Although previous studies have demonstrated that SLE is related to the imbalance of cells in the immune system, including B cells, T cells, and dendritic cells, etc., the mechanisms underlying SLE pathogenesis remain unclear. Therefore, effective and low side-effect therapies for SLE are lacking. Recently, mesenchymal stem cell (MSC) therapy for autoimmune diseases, particularly SLE, has gained increasing attention. This therapy can improve the signs and symptoms of refractory SLE by promoting the proliferation of Th2 and Treg cells and inhibiting the activity of Th1, Th17, and B cells, etc. However, MSC therapy is also reported ineffective in some patients with SLE, which may be related to MSC- or patient-derived factors. Therefore, the therapeutic effects of MSCs should be further confirmed. This review summarizes the status of MSC therapy in refractory SLE treatment and potential reasons for the ineffectiveness of MSC therapy from three perspectives. We propose various MSC modification methods that may be beneficial in enhancing the immunosuppression of MSCs in SLE. However, their safety and protective effects in patients with SLE still need to be confirmed by further experimental and clinical evidence.

• immunomodulation
• inefficacy
• mesenchymal stem cells
• modification
• systemic lupus erythematosus
• transplantation

• Cell therapy
• Fat depots
• H2AX
• MSC
• Mesenchymal stem cells
• Neural crest

• Adipose derived mesenchymal stromal cells
• Glycolysis
• Mitochondrial energy metabolism
• Proliferation
• Senescence

Overall, the human organism requires the production of ∼1 trillion new blood cells per day. Such goal is achieved via hematopoiesis occurring within the bone marrow (BM) under the tight regulation of hematopoietic stem and progenitor cell (HSPC) homeostasis made by the BM microenvironment. The BM niche is defined by the close interactions of HSPCs and non-hematopoietic cells of different origin, which control the maintenance of HSPCs and orchestrate hematopoiesis in response to the body’s requirements. The activity of the BM niche is regulated by specific signaling pathways in physiological conditions and in case of stress, including the one induced by the HSPC transplantation (HSCT) procedures. HSCT is the curative option for several hematological and non-hematological diseases, despite being associated with early and late complications, mainly due to a low level of HSPC engraftment, impaired hematopoietic recovery, immune-mediated graft rejection, and graft-versus-host disease (GvHD) in case of allogenic transplant. Mesenchymal stromal cells (MSCs) are key elements of the BM niche, regulating HSPC homeostasis by direct contact and secreting several paracrine factors. In this review, we will explore the several mechanisms through which MSCs impact on the supportive activity of the BM niche and regulate HSPC homeostasis. We will further discuss how the growing understanding of such mechanisms have impacted, under a clinical point of view, on the transplantation field. In more recent years, these results have instructed the design of clinical trials to ameliorate the outcome of HSCT, especially in the allogenic setting, and when low doses of HSPCs were available for transplantation.

• bone marrow niche
• hematopoietic (stem) cell transplantation
• hematopoietic stem and progenitor cell
• immunoregulation
• mesenchymal stromal cells

Mesenchymal stromal cells (MSCs) comprise a heterogeneous population of multipotent stromal cells that have gained attention for the treatment of irradiation-induced normal tissue toxicities due to their regenerative abilities. As the vast majority of studies focused on the effects of MSCs for photon irradiation-induced toxicities, little is known about the regenerative abilities of MSCs for particle irradiation-induced tissue damage or the effects of particle irradiation on the stem cell characteristics of MSCs themselves. MSC-based therapies may help treat particle irradiation-related tissue lesions in the context of cancer radiotherapy. As the number of clinical proton therapy centers is increasing, there is a need to decidedly investigate MSC-based treatments for particle irradiation-induced sequelae. Furthermore, therapies with MSCs or MSC-derived exosomes may also become a useful tool for manned space exploration or after radiation accidents and nuclear terrorism. However, such treatments require an in-depth knowledge about the effects of particle radiation on MSCs and the effects of MSCs on particle radiation-injured tissues. Here, the existing body of evidence regarding the particle radiobiology of MSCs as well as regarding MSC-based treatments for some typical particle irradiation-induced toxicities is presented and critically discussed.

• mesenchymal stem cells
• mesenchymal stromal cells
• normal tissue toxicities
• particle irradiation
• radiation accidents
• radiotherapy
• space irradiation
• stem cell therapy

• Conditioned medium
• Hypoxic preconditioning
• Immune regulation
• Ischemia–reperfusion injury
• Mesenchymal stem cells
• Microglia

• Animals
• Anti-Inflammatory Agents/metabolism
• Cell Hypoxia/drug effects
• Cell Line
• Cell Polarity/drug effects
• Cell Survival/drug effects
• Culture Media, Conditioned/pharmacology
• Exosomes/drug effects
• Exosomes/metabolism
• Exosomes/ultrastructure
• Glucose/deficiency
• Male
• Mesenchymal Stem Cells/drug effects
• Mesenchymal Stem Cells/pathology
• Mice
• Microglia/drug effects
• Microglia/pathology
• Mitochondria/drug effects
• Mitochondria/pathology
• Models, Biological
• Neuroprotective Agents/metabolism
• Oxygen
• Rats, Sprague-Dawley
• Reactive Oxygen Species/metabolism
• Reperfusion Injury/pathology
• Rats

• 81771707 National Natural Science Foundation of China
• 2042018kf0235 The Fundamental Research Funds for the Central Universities of China

Human mesenchymal stromal cells (hMSCs) have demonstrated, in various preclinical settings, consistent ability in promoting tissue healing and improving outcomes in animal disease models. However, translation from the preclinical model into clinical practice has proven to be considerably more difficult. One key challenge being the inability to perform in situ assessment of the hMSCs in continuous culture, where the accumulation of the senescent cells impairs the culture’s viability, differentiation potential and ultimately leads to reduced therapeutic efficacies. Histochemical \documentclass[12pt]{minimal} \usepackage{amsmath} \usepackage{wasysym} \usepackage{amsfonts} \usepackage{amssymb} \usepackage{amsbsy} \usepackage{mathrsfs} \usepackage{upgreek} \setlength{\oddsidemargin}{-69pt} \begin{document}$$\upbeta $$\end{document}β-galactosidase staining is the current standard for measuring hMSC senescence, but this method is destructive and not label-free. In this study, we have investigated alternatives in quantification of hMSCs senescence, which included flow cytometry methods that are based on a combination of cell size measurements and fluorescence detection of SA-\documentclass[12pt]{minimal} \usepackage{amsmath} \usepackage{wasysym} \usepackage{amsfonts} \usepackage{amssymb} \usepackage{amsbsy} \usepackage{mathrsfs} \usepackage{upgreek} \setlength{\oddsidemargin}{-69pt} \begin{document}$$\upbeta $$\end{document}β-galactosidase activity using the fluorogenic substrate, C\documentclass[12pt]{minimal} \usepackage{amsmath} \usepackage{wasysym} \usepackage{amsfonts} \usepackage{amssymb} \usepackage{amsbsy} \usepackage{mathrsfs} \usepackage{upgreek} \setlength{\oddsidemargin}{-69pt} \begin{document}$${_{12}}$$\end{document}12FDG; and autofluorescence methods that measure fluorescence output from endogenous fluorophores including lipopigments. For identification of senescent cells in the hMSC batches produced, the non-destructive and label-free methods could be a better way forward as they involve minimum manipulations of the cells of interest, increasing the final output of the therapeutic-grade hMSC cultures. In this work, we have grown hMSC cultures over a period of 7 months and compared early and senescent hMSC passages using the advanced flow cytometry and autofluorescence methods, which were benchmarked with the current standard in \documentclass[12pt]{minimal} \usepackage{amsmath} \usepackage{wasysym} \usepackage{amsfonts} \usepackage{amssymb} \usepackage{amsbsy} \usepackage{mathrsfs} \usepackage{upgreek} \setlength{\oddsidemargin}{-69pt} \begin{document}$$\upbeta $$\end{document}β-galactosidase staining. Both the advanced methods demonstrated statistically significant values, (r = 0.76, p \documentclass[12pt]{minimal} \usepackage{amsmath} \usepackage{wasysym} \usepackage{amsfonts} \usepackage{amssymb} \usepackage{amsbsy} \usepackage{mathrsfs} \usepackage{upgreek} \setlength{\oddsidemargin}{-69pt} \begin{document}$$\le $$\end{document}≤ 0.001 for the fluorogenic C\documentclass[12pt]{minimal} \usepackage{amsmath} \usepackage{wasysym} \usepackage{amsfonts} \usepackage{amssymb} \usepackage{amsbsy} \usepackage{mathrsfs} \usepackage{upgreek} \setlength{\oddsidemargin}{-69pt} \begin{document}$${_{12}}$$\end{document}12FDG method, and r = 0.72, p \documentclass[12pt]{minimal} \usepackage{amsmath} \usepackage{wasysym} \usepackage{amsfonts} \usepackage{amssymb} \usepackage{amsbsy} \usepackage{mathrsfs} \usepackage{upgreek} \setlength{\oddsidemargin}{-69pt} \begin{document}$$\le $$\end{document}≤ 0.05 for the forward scatter method), and good fold difference ranges (1.120–4.436 for total autofluorescence mean and 1.082–6.362 for lipopigment autofluorescence mean) between early and senescent passage hMSCs. Our autofluroescence imaging and spectra decomposition platform offers additional benefit in label-free characterisation of senescent hMSC cells and could be further developed for adoption for future in situ cellular senescence evaluation by the cell manufacturers.

Mesenchymal stromal cell (MSC)-based therapies are being actively investigated in various inflammatory disorders. However, functional variability among MSCs cultured in vitro will lead to distinct therapeutic efficacies. Until now, the mechanisms behind immunomodulatory functional variability in MSCs are still unclear.

We systemically investigated transcriptomic variations among MSC samples derived from multiple tissues to reveal their effects on immunomodulatory functions of MSCs. We then analyzed transcriptomic changes of MSCs licensed with INFγ to identify potential molecular mechanisms that result in distinct MSC samples with different immunomodulatory potency.

MSCs were clustered into distinct groups showing different functional enrichment according to transcriptomic patterns. Differential expression analysis indicated that different groups of MSCs deploy common regulation networks in response to inflammatory stimulation, while expression variation of genes in the networks could lead to different immunosuppressive capability. These different responsive genes also showed high expression variability among unlicensed MSC samples. Finally, a gene panel was derived from these different responsive genes and was able to regroup unlicensed MSCs with different immunosuppressive potencies.

This study revealed genes with expression variation that contribute to immunomodulatory functional variability of MSCs and provided us a strategy to identify candidate markers for functional variability assessment of MSCs.

The online version contains supplementary material available at 10.1186/s13287-020-02121-8.

• Expression variability
• Immune licensing
• Immunomodulatory potency
• MSC
• RNA-seq

• exosome
• mesenchymal stem cell
• radiation damage

• Animals
• Exosomes/physiology
• Humans
• Immunomodulation
• Mesenchymal Stem Cell Transplantation
• Mesenchymal Stem Cells/cytology
• Mice
• Neoplasms/radiotherapy
• Radiation Injuries/therapy
• Radiotherapy/adverse effects
• Rats
• Wound Healing

Adipose-derived stem cells (ADSCs) can maintain self-renewal and enhanced multidifferentiation potential through the release of a variety of paracrine factors and extracellular vesicles, allowing them to repair damaged organs and tissues. Consequently, considerable attention has increasingly been paid to their application in tissue engineering and organ regeneration. Here, we provide a comprehensive overview of the current status of ADSC preparation, including harvesting, isolation, and identification. The advances in preclinical and clinical evidence-based ADSC therapy for bone, cartilage, myocardium, liver, and nervous system regeneration as well as skin wound healing are also summarized. Notably, the perspectives, potential challenges, and future directions for ADSC-related researches are discussed. We hope that this review can provide comprehensive and standardized guidelines for the safe and effective application of ADSCs to achieve predictable and desired therapeutic effects.

• Age-associated diseases
• Cell senescence
• Mesenchymal stem cells
• Osteoarthritis
• Replication stress
• Senolytics

• P30 GM122732 NIGMS NIH HHS
• R33 AR076807 NIAMS NIH HHS
• R61 AR076807 NIAMS NIH HHS

Rare perivascular mesenchymal stromal cells (MSCs) with therapeutic properties have been identified in many tissues. Their rarity necessitates extensive in vitro expansion, resulting in spontaneous differentiation, cellular senescence and apoptosis, producing therapeutic products with variable quality and decreased potency. We previously demonstrated that A83-01, a transforming growth factor beta (TGF-β) receptor inhibitor, maintained clonogenicity and promoted the potency of culture-expanded premenopausal endometrial MSCs using functional assays and whole-transcriptome sequencing. Here, we compared the effects of A83-01 on MSCs derived from postmenopausal endometrium, menstrual blood, placenta decidua-basalis, bone marrow and adipose tissue. Sushi-domain-containing-2 (SUSD2⁺) and CD34⁺CD31⁻CD45⁻ MSCs were isolated. Expanded MSCs were cultured with or without A83-01 for 7 days and assessed for MSC properties. SUSD2 identified perivascular cells in the placental decidua-basalis, and their maternal origin was validated. A83-01 promoted MSC proliferation from all sources except bone marrow and only increased SUSD2 expression and prevented apoptosis in MSCs from endometrial-derived tissues. A83-01 only improved the cloning efficiency of postmenopausal endometrial MSCs (eMSCs), and expanded adipose tissue MSCs (adMSCs) underwent significant senescence, which was mitigated by A83-01. MSCs derived from bone marrow (bmMSCs) were highly apoptotic, but A83-01 was without effect. A83-01 maintained the function and phenotype in MSCs cultured from endometrial, but not other, tissues. Our results also demonstrated that cellular SUSD2 expression directly correlates with the functional phenotype.

• SUSD2
• adipose tissue
• apoptosis
• bone marrow
• clonogenicity
• endometrium
• menstrual fluid
• perivascular mesenchymal stromal cells
• placenta
• senescence

• Features
• Function
• Mesenchymal stem cells
• Rejuvenation strategy
• Senescence

• CD34+ HSCs
• EPO
• TGFβ1
• conditioned medium
• mesenchymal stromal cells

• mesenchymal stem cells
• inflammation
• immunotherapy
• stem-cell biotechnology
• tissue engineering

• Bioreactors
• Bone Marrow Cells
• Cell Culture Techniques/methods
• Cells, Cultured
• Cellular Reprogramming
• Cellular Reprogramming Techniques/methods
• Humans
• Immunomodulation/immunology
• Leukocytes, Mononuclear/immunology
• Lymphocytes/immunology
• Mesenchymal Stem Cells/immunology

• R01 EB012521 NIBIB NIH HHS
• R21 AI134116 NIAID NIH HHS
• R44 DK085766 NIDDK NIH HHS
• R44 HL128659 NHLBI NIH HHS

Mesenchymal stem cells (MSCs) are multilineage adult stem cells with considerable potential for cell‐based regenerative therapies. In vitro expansion changes their epigenetic and cellular properties, with a poorly understood impact on DNA damage response (DDR) and genome stability. We report here results of a transcriptome‐based pathway analysis of in vitro‐expanded human bone marrow‐derived mesenchymal stem cell (hBM‐MSCs), supplemented with cellular assays focusing on DNA double‐strand break (DSB) repair. Gene pathways affected by in vitro aging were mapped using gene ontology, KEGG, and GSEA, and were found to involve DNA repair, homologous recombination (HR), cell cycle control, and chromosomal replication. Assays for the recognition (γ‐H2AX + 53BP1 foci) and repair (pBRCA1 + γ‐H2AX foci) of X‐ray‐induced DNA DSBs in hBM‐MSCs show that over a period of 8 weeks of in vitro aging (i.e., about 10 doubling times), cells exhibit a reduced DDR and a higher fraction of residual DNA damage. Furthermore, a distinct subpopulation of cells with impaired DNA DSB recognition was observed. Several genes that participate in DNA repair by HR (e.g., Rad51, Rad54, BRCA1) show a 2.3‐ to fourfold reduction of their mRNA expression by qRT‐PCR. We conclude that the in vitro expansion of hMSCs can lead to aging‐related impairment of the recognition and repair of DNA breaks.

• BRCA1
• DNA repair
• cellular aging
• homologous recombination
• mesenchymal stem cells

• autophagy
• mesenchymal stem cells
• mitochondrial
• rejuvenation
• senescence
• telomere

Despite almost 50 years of research and over 20 years of preclinical and clinical studies, the question of curative potential of mesenchymal stem/stromal cells (MSCs) is still widely discussed in the scientific community. Non-reproducible treatment outcomes or even absence of treatment effects in comparison to control groups challenges the potential of these cells for routine application both in tissue engineering and in regenerative medicine. One of the reasons of such outcomes is non-standardized and often disadvantageous ex vivo manipulation of MSCs prior therapy. In most cases, clinically relevant cell numbers for MSC-based therapies can be only obtained by in vitro expansion of isolated cells. In this mini review, we will discuss point by point possible pitfalls in the production of human MSCs for cell therapies, without consideration of material-based applications. Starting with cell source, choice of donor and recipient, as well as isolation methods, we will then discuss existing expansion protocols (two-/three-dimensional cultivation, basal medium, medium supplements, static/dynamic conditions, and hypoxic/normoxic conditions) and influence of these strategies on the cell functionality after implantation. The role of potency assays will also be addressed. The final aim of this mini review is to illustrate the heterogeneity of current strategies for gaining MSCs for clinical applications with their strengths and weaknesses. Only a careful consideration and standardization of all pretreatment processes/methods for the different applications of MSCs will ensure robust and reproducible performance of these cell populations in the different experimental and clinical settings.

• bioreactor
• donor variability
• expansion protocols
• mesenchymal stem/stromal cell
• potency

• MSC senescence
• in vitro aging
• in vivo aging
• mesenchymal stem/stromal cells (MSC)
• rejuvenating strategies

• Aging/physiology
• Cell Proliferation
• Cells, Cultured
• Cellular Senescence/physiology
• Humans
• Mesenchymal Stem Cells/metabolism
• Mesenchymal Stem Cells/physiology

• 5x1000 Italian Health Ministry
• RFO Bologna University
• Ricerca Corrente IRCCS Istituto Ortopedico Rizzoli

• autologous stem cell ovarian transplant
• follicular rescue
• mobilization
• ovarian rejuvenation
• premature ovarian insufficiency
• stem cells

• Animals
• Clinical Trials as Topic/methods
• Disease Models, Animal
• Female
• Humans
• Paracrine Communication/immunology
• Primary Ovarian Insufficiency/immunology
• Primary Ovarian Insufficiency/therapy
• Stem Cell Transplantation/methods
• Stem Cells/immunology
• Treatment Outcome