scholarly journals Human amniotic mesenchymal stem cells combined with PPCNg facilitate injured endometrial regeneration

2022 ◽  
Vol 13 (1) ◽  
Author(s):  
Jiayue Huang ◽  
Wenwen Zhang ◽  
Jie Yu ◽  
Yating Gou ◽  
Nizhou Liu ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Fluorescence Imaging ◽  
Surface Markers ◽  
Multilineage Differentiation ◽  
Differentiation Potential ◽  
Qrt Pcr ◽  
Amniotic Mesenchymal Stem Cells ◽  
Endometrial Regeneration

Abstract Background Caused by the injury to the endometrial basal layer, intrauterine adhesions (IUA) are characterized by uterine cavity obliteration, leading to impaired fertility. Human amniotic mesenchymal stem cells (hAMSCs) have the potential to promote endometrial regeneration mainly through paracrine ability. PPCNg is a thermoresponsive biomaterial consisted of Poly (polyethylene glycol citrate-co-N-isopropylacrylamide) (PPCN) mixed with gelatin, which has been reported as a scaffold for stem cell transplantation. This study aims to investigate the therapeutic effect of hAMSCs combined with PPCNg transplantation in promoting the regeneration of injured endometrium. Methods hAMSCs were cultured in different concentrates of PPCNg in vitro, and their proliferation, apoptosis and cell cycle were examined by CCK-8 assay and flow cytometry. Immunofluorescence was used to determine the MSCs specific surface markers. The expression of pluripotent genes was analyzed by qRT-PCR. The multiple-lineage differentiation potential was further evaluated by detecting the differentiation-related genes using qRT-PCR and specific staining. The Sprague–Dawley (SD) rat IUA model was established with 95% ethanol. hAMSCs combined with PPCNg were transplanted through intrauterine injection. The retention of DiR-labeled hAMSCs was observed by vivo fluorescence imaging. The endometrium morphology was assessed using hematoxylin and eosin (H&E) and Masson staining. Immunohistochemistry staining was performed to detect biomarkers related to endometrial proliferation, re-epithelialization, angiogenesis and endometrial receptivity. The function of regenerated endometrium was evaluated by pregnancy tests. Results hAMSCs maintained normal cell proliferation, apoptosis and cell cycle in PPCNg. Immunofluorescence and qRT-PCR showed that hAMSCs cultured in PPCNg and hAMSCs cultured alone expressed the same surface markers and pluripotent genes. hAMSCs exhibited normal multilineage differentiation potential in PPCNg. Vivo fluorescence imaging results revealed that the fluorescence intensity of hAMSCs combined with PPCNg intrauterine transplantation was stronger than that of direct hAMSCs intrauterine transplantation. Histological assays showed the increase in the thickness of endometrial and the number of endometrial glands, and the remarkably decrease in the fibrosis area in the PPCNg/hAMSCs group. The expressions of Ki-67, CK7, CK19, VEGF, ER and PR were significantly increased in the PPCNg/hAMSCs group. Moreover, the number of implanted embryos and pregnancy rate were significantly higher in the PPCNg/hAMSCs group than in the hAMSCs group. Conclusions PPCNg is suitable for growth, phenotype maintenance and multilineage differentiation of hAMSCs. hAMSCs combined with PPCNg intrauterine transplantation can facilitate the regeneration of injured endometrium by improving utilization rates of hAMSCs, and eventually restore reproductive capacity.

scholarly journals Characterization of Human Mesenchymal Stem Cells from Different Tissues and Their Membrane Encasement for Prospective Transplantation Therapies

2019 ◽  
Vol 2019 ◽  
pp. 1-13 ◽  
Author(s):  
Eva Schmelzer ◽  
Daniel T. McKeel ◽  
Jörg C. Gerlach
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Growth Factors ◽  
Cell Fusion ◽  
Human Mesenchymal Stem Cells ◽  
Surface Markers ◽  
Multilineage Differentiation ◽  
Differentiation Potential ◽  
Donor Cells ◽  
Different Tissues

Human mesenchymal stem cells can be isolated from various organs and are in studies on therapeutic cell transplantation. Positive clinical outcomes of transplantations have been attributed to both the secretion of cytokines and growth factors as well as the fusion of donor cells with that of the host. We compared human mesenchymal stem cells from six different tissues for their transplantation-relevant potential. Furthermore, for prospective allogenic transplantation we developed a semipermeable hollow-fiber membrane enclosure, which would prevent cell fusion, would provide an immune barrier, and would allow for easy removal of donor cells from patients after recovery. We investigated human mesenchymal stem cells from adipose tissue, amniotic tissue, bone marrow, chorionic tissue, liver, and umbilical cord. We compared their multilineage differentiation potential, secretion of growth factors, and the expression of genes and surface markers. We found that although the expression of typical mesenchymal stem cell-associated gene THY1 and surface markers CD90 and CD73 were mostly similar between mesenchymal stem cells from different donor sites, their expression of lineage-specific genes, secretion of growth factors, multilineage differentiation potential, and other surface markers were considerably different. The encasement of mesenchymal stem cells in fibers affected the various mesenchymal stem cells differently depending on their donor site. Conclusively, mesenchymal stem cells isolated from different tissues were not equal, which should be taken into consideration when deciding for optimal sourcing for therapeutic transplantation. The encasement of mesenchymal stem cells into semipermeable membranes could provide a physical immune barrier, preventing cell fusion.

scholarly journals Heterogeneity of mesenchymal stem cells: characterization and application in cell therapy

STEMedicine ◽  
2022 ◽  
Vol 3 (1) ◽  
pp. e109
Author(s):  
Xingzhi Liu ◽  
Zhihua Zhao ◽  
Zhe Zhao ◽  
Zhongjuan Xu ◽  
Junjun Cao ◽  
...  
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Regenerative Medicine ◽  
Cell Therapy ◽  
Clinical Application ◽  
Surface Markers ◽  
Multilineage Differentiation ◽  
Differentiation Potential ◽  
Approaches To Study ◽  
Major Factors

Mesenchymal stem cells (MSCs) have shown great potentials in regenerative medicine for their low immunogenicity, multilineage differentiation potential, and extensive sources. However, the heterogeneity of MSCs limits their clinical application and industrial prospects. In this review, we introduced the heterogeneity of MSCs in terms of their applications, sources, functions, and surface markers; discussed the major factors leading to the heterogeneity in MSCs; summarized the main approaches to study the MSC heterogeneity, and addressed the clinical challenges resulting from heterogeneity. Finally, we proposed the strategies that might be used to purify the MSCs and to eliminate the heterogeneity of MSCs for their standardized production and reliable clinical application.

Differential Expression of MicroRNA-3148 in Patients with Osteoporosis and Its Impacts on the Osteogenic Differentiation of Rat Bone Marrow Mesenchymal Stem Cells

2021 ◽  
Vol 11 (5) ◽  
pp. 957-962
Author(s):  
Ainiwaerjiang Damaola ◽  
Maerdan Aierken ◽  
Mieralimu Muertizha ◽  
Abudouaini Abudoureheman ◽  
Haishan Lin ◽  
...  
Keyword(s):  
Stem Cells ◽  
Bone Marrow ◽  
Mesenchymal Stem Cells ◽  
Osteogenic Differentiation ◽  
Serum Samples ◽  
Alizarin Red ◽  
Differentiation Potential ◽  
Qrt Pcr ◽  
Marrow Mesenchymal Stem Cells ◽  
Rat Bone

We aimed to explore the effects of rat bone marrow mesenchymal stem cells (BMSCs) on osteogenic differentiation via analyzing miR-3148 expression in patients with osteoporosis. Realtime quantitative PCR was conducted for assessing microRNA-3148 expression. BMSCs from SD rats were transfected with microRNA-3148 mimics and microRNA-3148 inhibitor via liposomal trans-fection method utilizing Lipo2000, followed by analysis of microRNA-3148 level. After 10-days of osteogenic differentiation induction, alkaline phosphatase (ALP) staining and alizarin red (ARS) staining were done to investigate the osteogenic differentiation potential. Simultaneously, qRT-PCR measured the expression of osteogenesis marker genes (BMP and Runx2) in each group. qRT-PCR analysis revealed a high expression of miR-3148 in the bone tissue and the serum samples from patients with osteoporosis in comparison with healthy individuals. In addition, miRNA-3148 mimics could retard the osteogenic differentiation of BMSCs, while microRNA-3148 inhibitor could prompt the procedure. MicroRNA-3148 was highly expressed in the skeletal tissues and the serum samples from patients with osteoporosis and it could restrain the differentiation of BMSCs into osteoblasts, suggesting that it might be a novel therapeutic target for treating osteoporosis.

Purinergic Stimulation of Human Bone Marrow-Derived Mesenchymal Stem Cells Modulate Their Function and Differentiation Potential.

Blood ◽  
2010 ◽  
Vol 116 (21) ◽  
pp. 3848-3848
Author(s):  
Marilena Ciciarello ◽  
Valentina Salvestrini ◽  
Davide Ferrari ◽  
Sara Gulinelli ◽  
Roberta Zini ◽  
...  
Keyword(s):  
Stem Cells ◽  
Bone Marrow ◽  
Cell Proliferation ◽  
Mesenchymal Stem Cells ◽  
Transcription Factor ◽  
Human Bone ◽  
Differentiation Potential ◽  
Qrt Pcr ◽  
Differentiated Cells

Abstract Abstract 3848 Introduction: Human bone marrow derived Mesenchymal Stem Cells (hMSCs) are adult multipotent cells. hMSCs differentiate in vitro and in vivo into several tissue lineages originating from the three germinal layers making them attractive candidates for bioengineering and cellular therapy. Thus, it seems of great relevance to search putative messengers and signalling able to modulate their proliferation and differentiation. Nucleotides triphosphates are extracellular messengers binding to specific receptors (P2Rs) that modulate cell functions depending on the cell type. Controversial information is available on P2 expression and activity in hMSCs. Methods and Results: Here we found that hMSCs expressed several P2R subtypes. hMSCs were very resistant to the cytotoxic effects of high concentrations of ATP, as demonstrated by the lack of morphological and mitochondrial changes or release of intracellular markers of cell death. Gene expression profiling revealed that ATP treatment down-regulated cell proliferation and up-regulated cell migration genes in hMSCs. Functional studies confirmed the inhibitory activity of ATP on proliferation and clonogenic ability of hMSCs. Furthermore, ATP potentiated the chemotactic response of hMSCs to the chemokine CXCL12, and increased their spontaneous migration. In vivo, xenotransplant experiments showed that the homing capacity of hMSCs to murine bone marrow was increased by ATP pre-treatment. Moreover, ATP increased pro-inflammatory cytokines production (IL-2, IFN-g, IL-12p70), while decreased secretion of the anti-inflammatory cytokine IL-10. This finding was associated with the reduced ability of ATP-treated hMSC of inhibiting T-cell proliferation. Microarrays data suggested that several genes implicated in hMSC differentiation can be modulated by ATP treatment. To further investigate this issue, hMSCs cells were cultured under adipogenic or osteogenic conditions and were transiently exposed to ATP before starting differentiation or continuously exposed to ATP for the first 3 days of differentiation induction. We demonstrated that adipogenesis-related accumulation of lipids, analyzed by Oil red O staining, was more evident in ATP treated cultures. Furthermore, quantitative real time PCR (qRT-PCR) assay showed that mRNA expression of PPARg, a transcription factor early up-regulated during adipogenesis, was significantly increased in hMSCs differentiated cells treated with ATP. In osteogenic condition, analysis of mineralized area through Alizarin Red staining, indicated that ATP treatment enhanced the extent of mineralization compared to untreated control. The expression of RUNX2, a key transcription factor in osteogenesis, analyzed by qRT-PCR in differentiated cells confirmed data obtained in Alizarin-based assay. Conclusions: These data demonstrated that purinergic signalling modulates biological functions and differentiation potential of hMSCs. Disclosures: No relevant conflicts of interest to declare.

scholarly journals Mesenchymal stem cells – a historical overview

2020 ◽  
Vol 8 (2) ◽  
pp. 83-87
Author(s):  
Katarzyna Stefańska ◽  
Rut Bryl ◽  
Lisa Moncrieff ◽  
Nelson Pinto ◽  
Jamil A. Shibli ◽  
...  
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Cellular Therapy ◽  
Osteogenic Potential ◽  
Experimental Medicine ◽  
Cancer Tissue ◽  
Multilineage Differentiation ◽  
Differentiation Potential ◽  
Clinical Utilization ◽  
History Of

AbstractMesenchymal stem cells are currently one of the most extensively studied topics in experimental medicine, given their unique properties in terms of immunomodulation, multiple factors secretion and homing to injured tissue sites. Such characteristics were proven to be invaluable in various disease management treatments, for example in cancer, tissue regeneration or immunologic/inflammatory-related disorders. MSCs were first isolated from bone marrow in 1960-1970’s and were characterized as cells with fibroblastoid shape and osteogenic potential, which form clonogenic colonies (CFU-F – colony-forming unit-fibroblast). Nowadays the term ‘mesenchymal stem cells’ is used in regards to all of the cells meeting minimal criteria published in 2006 by the International Society for Cellular Therapy, however the name ‘mesenchymal stromal cells’ has been suggested to be more appropriate. Regardless of the name controversy, these cells exhibit multilineage differentiation potential, self-renewal ability, adhere to plastic and express specific surface antigens. In 2011 the first commercial product based on MSCs was developed and many more are expected to emerge. This review focuses on a historical perspective concerning studies on MSCs, controversies regarding their name and their characteristics and clinical utilization.Running title: The history of mesenchymal stem cells

scholarly journals A novel method for maintaining the stability of freshly cultured Mesenchymal stem cells in clinical grade injection ready state without cryopreservation

2021 ◽  
Vol 6 (1) ◽  
Author(s):  
Mandana Mohyeddin Bonab ◽  
Fatemeh Talebian ◽  
Aida Borzabadi ◽  
Vahideh Nasr ◽  
Azam Abedi Kooshlshahi ◽  
...  
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Human Plasma ◽  
Room Temperature ◽  
Growth Potential ◽  
Surface Markers ◽  
Cell Surface Markers ◽  
Clinical Grade ◽  
Differentiation Potential ◽  
Novel Method

Abstract Background Mesenchymal Stem Cells (MSCs) are multipotent cells with low immuonogenecity, and dynamic tissue repair potential, which explains the overwhelming attention they have attracted in regenerative therapy. One notable challenge in MSCs therapy is the bench to bed timeline of freshly cultured MSCs; it does not exceed 24 h. For use after 24 h, MSC need to be cryopreserved - which can preserve the cells for years - but it is a costly and damaging process. Here we introduce a method to extend the bench to bed lifetime of MSCs up to 4 days without the high cost and cell damaging effects of cryopreservation. Our method is based on preserving the MSCs in human plasma. Methods MSCs of 12 tissue samples - 4 adipose, 4 bone marrow and 4 Wharton’s jelly- were cultured and expanded in standard conditions. Cells harvested from passage 2 or 3 were washed, centrifuged, pelleted, and re-suspended in human plasma. Cell suspensions were refrigerated (5 ± 3 °C) or stored at room temperature (22 ± 3 °C) in a sterile, temperature controlled room. During the next 7 days, two tubes (one from each group) were examined every 24 h to assess MSCs viability and growth potential. On day 3, we assessed MSC cell surface markers and its differentiation potential to adipocyte and osteocyte tissues. Results were analyzed by computing the overall mean and applying the independent-samples t-test to those means. Results The sample means for both cell expansion and cell viability were compared between the two “refrigerator” and “room temperature” groups. Although there was a gradual decrease in cell growth potential between the cells stored for 1 day to those stored for 7 days, we show more than 80% of the cells remain alive for up to 4 days of storage in both groups. The cells reached 80% confluency in under 20 days for all samples stored up to 4 days. No significant differences were observed between the two groups (room temperature and refrigerator stored). The differentiation potential to adipocyte and osteocyte tested on day 3 were positive in all samples. The analysis of cell surface markers tested on day 3 were positive for CD90, CD105, CD73 and negative for CD34, CD45 and HLA-DR. Conclusion We present a method of MSC culture medium using human plasma that can preserve their viability and growth potential for up to 4 days in both room and refrigerator temperatures without losing their stemness characteristics (we recommend use of 5 ± 3 °C). This novel method will allow rapid expansion and therapeutic use of MSCs. Since the cells can be maintained in clinical grade, injection ready state for several days, they can be transported across the globe.

scholarly journals The effect of titanium carbide used in implants on stemness and senescence of mouse bone marrow derived mesenchymal stem cells

2020 ◽  
Vol 42 (5) ◽  
pp. 537-546
Author(s):  
Roya Salehi ◽  
Roya Valizadeh ◽  
Akram Farzandi ◽  
Effat Alizadeh
Keyword(s):  
Cell Cycle ◽  
Stem Cells ◽  
Bone Marrow ◽  
Mesenchymal Stem Cells ◽  
Titanium Carbide ◽  
Mtt Assay ◽  
Mouse Bone Marrow ◽  
Qrt Pcr ◽  
Colony Forming Assay ◽  
Beta Galactosidase

Background: The aim of the present study was to determine the effect of Titanium carbide nanoparticles on stemness and senescence markers of mouse bone marrow derived mesenchymal stem cells. Methods: After isolation of mesenchymal stem cells from mouse’s bone marrow, their surface markers were studied using flowcytometry. MTT assay, cell cycle analysis; colony forming assay, and senescent beta galactosidase staining were performed for cells treated with titanium carbide nanoparticles. Then, the expression of OCT-4 and Nanog genes were studied by qRT-PCR in titanium carbide treated mouse’s bone marrow mesenchymal stem cells (mBMSCs). Results: All mBMSCs showed spindle shaped morphology. Their identity was confirmed by flow cytometry of stained cells for CD11b, CD90, CD45, and CD44 markers. The MTT assay results showed that titanium carbide effect is time-dependent and no cytotoxic effects were observed in short treatment times. But toxic effects were observed after 72 or 144 hours of post treatment with doses range from 0.1 to 1 mM (P<0.05). Besides, cell cycle study detected more cell populations in G0/G1 and less percentage in S phase. The colony forming assay in treated cells exhibited smaller size of colonies. The beta galactosidase staining of treated cells demonstrated more positive cells (more aged cells). Finally, qRT-PCR showed significant down regulation of OCT-4 and Nanog (p<0.05). Conclusion: Titanium carbide as an implant material could affect the viability, stemness and senescence of mBMSCs in negative manner.

scholarly journals More to Explore; The Mesenchymal Stem Cells (MSCs) Major Tissue Sources, Known Surface Markers, and Its Immunomodulation properties

2021 ◽  
pp. 85-97
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Immune Modulation ◽  
Molecular Mechanisms ◽  
Cell Types ◽  
Surface Markers ◽  
Differentiation Potential ◽  
Disease Therapy ◽  
Multipotent Progenitor Cells ◽  
Mesodermal Origin

Mesenchymal stem cells (MSCs) are currently available for a range of applications and have become a good material for regenerative medicine, tissue engineering, and disease therapy. MSCs are self-renewing, multipotent progenitor cells with multilineage potential to differentiate into cell types of mesodermal origin, such as adipocytes, osteocytes, and chondrocytes, and exert potent immunosuppressive potentials. In the present review, we highlight the currently reported variations in the differentiation potential of MSCs from different tissue sources, the minimal criteria to define MSCs from various tissue environments, and provide a detailed description of MSCs surface markers. Furthermore, MSC's immunomodulatory features secrete cytokines and immune receptors which regulate the microenvironment in the host tissue also revisits in detail. We propose that there are likely more sources of MSCs waiting to be discovered. We need to Standardize MSCs characterization by selecting markers for isolation, cellular and molecular mechanisms involved in MSC-mediated immune modulation, and other functionalities of MSCs should be characterized prior to use in clinical applications.

scholarly journals The differentiation potential of adipose tissue-derived mesenchymal stem cells into cell lineage related to male germ cells

2018 ◽  
Vol 70 (1) ◽  
pp. 160-168 ◽  
Author(s):  
P. Bräunig ◽  
W.G. Glanzner ◽  
V.B. Rissi ◽  
P.B.D. Gonçalves
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Adipose Tissue ◽  
Cell Lineage ◽  
Gene Marker ◽  
Surface Markers ◽  
Differentiation Potential ◽  
Fat Depots ◽  
Reliable Source ◽  
Stromal Stem Cells

ABSTRACT The adipose tissue is a reliable source of Mesenchymal stem cells (MSCs) showing a higher plasticity and transdifferentiation potential into multilineage cells. In the present study, adipose tissue-derived mesenchymal stem cells (AT-MSCs) were isolated from mice omentum and epididymis fat depots. The AT-MSCs were initially compared based on stem cell surface markers and on the mesodermal trilineage differentiation potential. Additionally, AT-MSCs, from both sources, were cultured with differentiation media containing retinoic acid (RA) and/or testicular cell-conditioned medium (TCC). The AT-MSCs expressed mesenchymal surface markers and differentiated into adipogenic, chondrogenic and osteogenic lineages. Only omentum-derived AT-MSCs expressed one important gene marker related to male germ cell lineages, after the differentiation treatment with RA. These findings reaffirm the importance of adipose tissue as a source of multipotent stromal-stem cells, as well as, MSCs source regarding differentiation purpose.

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