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Biomed Res Int ; 2021: 4604856, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34527737


IFN-γ licensing to mesenchymal stem cells (MSCs) is applied to enhance the therapeutic potential of MSCs. However, although the features of MSCs are affected by several stimuli, little information is available on changes to the therapeutic potential of IFN-γ-licensed differentiated MSCs during xenogeneic applications. Therefore, the present study is aimed at clarifying the effects of adipogenic/osteogenic differentiation and IFN-γ licensing on the in vitro immunomodulatory and migratory properties of porcine bone marrow-derived MSCs in xenogeneic applications using human peripheral blood mononuclear cells (PBMCs). IFN-γ licensing in differentiated MSCs lowered lineage-specific gene expression but did not affect MSC-specific cell surface molecules. Although indoleamine 2,3 deoxygenase (IDO) activity and expression were increased after IFN-γ licensing in undifferentiated MSCs, they were reduced after differentiation. IFN-γ licensing to differentiated MSCs elevated the reduced IDO expression in differentiated MSCs; however, the increase was not sufficient to reach to the level achieved by undifferentiated MSCs. During a mixed lymphocyte reaction with quantification of TNF-α concentration, proliferation and activation of xenogeneic PBMCs were suppressed by undifferentiated MSCs but inhibited to a lesser extent by differentiated MSCs. IFN-γ licensing increasingly suppressed proliferation of PBMCs in undifferentiated MSCs but it was incapable of elevating the reduced immunosuppressive ability of differentiated MSCs. Migratory ability through a scratch assay and gene expression study was reduced in differentiated MSCs than their undifferentiated counterparts; IFN-γ licensing was unable to enhance the reduced migratory ability in differentiated MSCs. Similar results were found in a Transwell system with differentiated MSCs in the upper chamber toward xenogeneic PBMCs in the lower chamber, despite IFN-γ licensing increased the migratory ability of undifferentiated MSCs. Overall, IFN-γ licensing did not enhance the reduced immunomodulatory and migratory properties of differentiated MSCs in a xenogeneic application. This study provides a better understanding of the ways in which MSC therapy can be applied.

Interferon gama/metabolismo , Transplante de Células-Tronco Mesenquimais/métodos , Células-Tronco Mesenquimais/metabolismo , Animais , Medula Óssea/metabolismo , Células da Medula Óssea/metabolismo , Diferenciação Celular/efeitos dos fármacos , Movimento Celular/efeitos dos fármacos , Proliferação de Células/efeitos dos fármacos , Células Cultivadas , Técnicas de Cocultura , Citocinas/metabolismo , Xenoenxertos/metabolismo , Humanos , Imunomodulação/efeitos dos fármacos , Interferon gama/fisiologia , Células-Tronco Mesenquimais/fisiologia , Osteogênese/efeitos dos fármacos , Suínos , Fator de Necrose Tumoral alfa/metabolismo
Biomed Res Int ; 2021: 5540877, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34337022


Mesenchymal stem cells (MSCs) are valuable candidates in tissue engineering and stem cell-based therapy. Traditionally, MSCs derived from various tissues have been successfully expanded in vitro using adherent culture plates commonly called as monolayer two-dimensional (2D) cultures. Recently, many studies demonstrated that stemness and multilineage differentiation potential could be enhanced to greater extent when MSCs are cultured as suspended aggregates by means of three-dimensional (3D) culturing techniques. However, there are limited reports on changed mitochondrial metabolism on 3D spheroid formation of MSCs. Therefore, the present study was aimed at investigating the stemness, differentiation potential, and mitochondrial metabolism capacity of 3D dental pulp-derived MSC (DPSC) spheroids in comparison to monolayer cultured DPSCs. We isolated dental pulp-derived MSCs (DPSCs) and successfully developed a 3D culture system which facilitated the formation of MSC spheroids. The cell aggregation was observed after 2 hours, and spheroids were formed after 24 hours and remained in shape for 72 hours. After spheroid formation, the levels of pluripotent markers increased along with enhancement in adipogenic and osteogenic potential compared to 2D cultured control cells. However, decreased proliferative capacity, cell cycle arrest, and elevated apoptosis rate were observed with the time course of the 3D culture except for the initial 24-hour aggregation. Furthermore, oxygen consumption rates of living cells decreased with the time course of the aggregation except for the initial 24 hours. Overall, our study indicated that the short-term 3D culture of MSCs could be a suitable alternative to culture the cells.

Diferenciação Celular , Polpa Dentária/citologia , Células-Tronco Mesenquimais/citologia , Mitocôndrias/metabolismo , Células-Tronco Pluripotentes/citologia , Esferoides Celulares/citologia , Adipogenia , Apoptose , Biomarcadores/metabolismo , Técnicas de Cultura de Células , Ciclo Celular , Proliferação de Células , Células Cultivadas , Humanos , Células-Tronco Mesenquimais/metabolismo , Osteogênese , Consumo de Oxigênio , Células-Tronco Pluripotentes/metabolismo , Esferoides Celulares/metabolismo
Int J Mol Sci ; 22(4)2021 Feb 17.
Artigo em Inglês | MEDLINE | ID: mdl-33671303


In the present era, infertility is one of the major issues which restricts many couples to have their own children. Infertility is the inability to achieve a clinical pregnancy after regular unprotected sexual intercourse for the period of one year or more. Various factors including defective male or female germ cell development, unhealthy and improper lifestyles, diseases like cancer and associated chemo-or-radiation therapies, congenital disorders, etc., may be responsible for infertility. Therefore, it is highly important to understand the basic concepts of germ cell development including primordial germ cell (PGC) formation, specification, migration, entry to genital ridges and their molecular mechanisms, activated pathways, paracrine and autocrine signaling, along with possible alteration which can hamper germ cell development and can cause adversities like cancer progression and infertility. Knowing all these aspects in a proper way can be very much helpful in improving our understanding about gametogenesis and finding possible ways to cure related disorders. Here in this review, various aspects of gametogenesis especially female gametes and relevant factors causing functional impairment have been thoroughly discussed.

Células Germinativas/patologia , Animais , Carcinogênese/patologia , Epigênese Genética , Feminino , Humanos , Neoplasias Embrionárias de Células Germinativas/complicações , Neoplasias Embrionárias de Células Germinativas/diagnóstico , Neoplasias Embrionárias de Células Germinativas/genética , Neoplasias Embrionárias de Células Germinativas/patologia , Oócitos/citologia
Int J Med Sci ; 18(5): 1259-1268, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-33526987


Background: Multipotent and immune privileged properties of mesenchymal stem cells (MSCs) were investigated for the treatment of various clinical diseases. For the years, many researches into the animal studies evaluated human stem cell therapeutic capacity related to the regenerative medicine. However, there were limited reports on immune privileged properties of human MSCs in animal studies. The present study investigated hematological and biochemical parameter and lymphocyte subset in mini-pigs following human MSCs transplantation as a means of validation of reliability that influence the animal test results. Methods: The miniature pigs were transplanted with human MSCs seeded with scaffold. After transplantation, all animals were evaluated by CBC, biochemistry and lymphocyte subset test. After 9 weeks, all pigs were sacrificed and organs were histologically analyzed. Results: CBC test showed that levels of RBC were decreased and reticulocyte, WBC and neutrophil were increased in transient state initially after transplantation, but returned to normal value. The proportion of B lymphocyte and cytotoxic T cell were also initially enhanced within the normal range temporarily. The female and male miniature pigs showed normal ranges for blood chemistry assessments. During the 9 weeks post-operative period, the animals showed a continuous increase in body weight and length. Furthermore, no abnormal findings were observed from the histological analysis of sacrificed pigs. Conclusions: Overall, miniature pigs transplanted with human MSCs seeded with scaffold were found to have physiologically similar results to normal animals. This result might be a reliable indicator of the animal experiments using miniature pigs with human MSCs.

Privilégio Imunológico , Transplante de Células-Tronco Mesenquimais/métodos , Células-Tronco Mesenquimais/imunologia , Porco Miniatura/imunologia , Animais , Contagem de Células Sanguíneas , Feminino , Humanos , Masculino , Modelos Animais , Medicina Regenerativa/métodos , Reprodutibilidade dos Testes , Suínos , Tecidos Suporte , Transplante Heterólogo