scholarly journals Optimization of Tenocyte Lineage-related Factors from Tonsil-derived Mesenchymal Stem Cells using Response Surface Methodology

2018 ◽  
Author(s):  
Seung Yeol Lee ◽  
Hyang Kim ◽  
Sang-Jin Shin ◽  
Soon-Sun Kwon
Keyword(s):  
Stem Cells ◽  
Bone Marrow ◽  
Mesenchymal Stem Cells ◽  
Transforming Growth Factor Beta ◽  
Mononuclear Cells ◽  
Prediction Models ◽  
Transforming Growth Factor ◽  
Related Factors ◽  
Tenogenic Differentiation ◽  
Potential Factors

AbstractResearchers should consider various potential factors that affect tenogenic differentiation of mesenchymal stem cells (MSCs); however, this requires numerous experimental settings, which are associated with high cost and time. We aimed to assess the differential effects of transforming growth factor beta 3 (TGF-β3) on the tenogenesis of tonsil-derived MSCs (T-MSCs) and bone marrow-derived MSCs (BM-MSCs) using design of experiments (DoE). Bone marrow and tonsillar tissue was collected from four patients; mononuclear cells were separated and treated with 5 and 10 ng/mL of TGF-β3 with vehicle control. A full-factorial experimental design with a categorical factor of 0 was employed to study the effect of tension based on T-MSCs. Eighty-four trials were utilized, fitted with RSM, and then used to obtain mathematical prediction models. Exposure of T-MSCs and BM-MSCs to TGF-β3 increased the expression of scleraxis (SCX), tenomodulin (TNMD), decorin, collagen I, and tenascin C. Expression of most of these factors reached a maxima after 2–3 days of treatment. Considering all of the tenocyte lineage-related factors that were assessed, the predicted value of the factors from T-MSCs was significantly induced at 2.7 ng/mL of TGF-β3 during 2.5-day culture, whereas the predicted value of the factors from BM-MSCs was significantly induced during 2.3-day culture, regardless of TGF-β3 concentration. This study demonstrated that tenogenic differentiation of T-MSCs and BM-MSCs under TGF-β3 stimulation showed a similar culture time for peak expression of tenocyte-related mRNAs using RSM. This study suggests the potential of using the DoE approach for optimization of the culture protocol for tenogenesis of MSCs.

scholarly journals Optimization of Tenocyte Lineage-Related Factors from Mesenchymal Stem Cells

2018 ◽  
Vol 3 (3) ◽  
pp. 2473011418S0031
Author(s):  
Seung Yeol Lee ◽  
Hyang Kim ◽  
Soon-Sun Kwon
Keyword(s):  
Stem Cells ◽  
Bone Marrow ◽  
Mesenchymal Stem Cells ◽  
Transforming Growth Factor Beta ◽  
Mononuclear Cells ◽  
Prediction Models ◽  
Transforming Growth Factor ◽  
Related Factors ◽  
Differentiation Potential ◽  
Tenogenic Differentiation

Category: Basic Sciences/Biologics Introduction/Purpose: Researchers should consider various potential factors that affect tenogenic differentiation of MSCs. Numerous experimental settings are associated with high cost and time. Response surface methodology (RSM), a component in the design of experiments (DOE), is gaining recognition as a powerful tool in optimizing conditions for the production of industrially important products such as chemicals and enzymes. The purpose of this study was to access the differential effects of transforming growth factor beta 3 (TGF-β3) on the tenogenesis of tonsil-derived mesenchymal stem cells (T-MSCs) and bone marrow-derived mesenchymal stem cells (BM-MSCs) using RSM. Methods: Bone marrow was collected from four patients (mean age: 79.0±2.2) and mononuclear cells were separated. The tonsillar tissues were collected from four patients (mean age: 7.6±0.6). After isolation of MSCs, they were treated with 5ng/ml and 10ng/ml of TGF-β3 with vehicle control. The full-factorial experimental design was employed to study the effect of tension based on T-MSCs. The design was composed of three levels being coded as -1, 0 and +1 and a total of 18 runs were carried out in duplicates to optimize the level of chosen variables, such as days and amount. A total of 84 trials were utilized and fitted with RSM; they were then used to obtain mathematical prediction models. Results: Exposure of TGF-β3 to T-MSCs and BM-MSCs resulted in an increase in the expression of SCX, TNMD, decorin, collagen I, and tenacin C. Most tenocyte lineage-related factors from T-MSCs and BM-MSCs presented a maximum increase in 2- 3 day treatment. Considering all of tenocyte lineage-related factors that we assessed, the predicted value of the factors was significantly induced at 2.7 ng/mL of TGF-β3 (p < 0.001) on 2.5-day culture (p = 0.001). (Fig A) Considering all of tenocyte lineage-related factors that we assessed, the predicted value of the factors was significantly induced on 2.3-day culture (p = 0.004) regardless of the concentration of TGF-β3. (Fig B) Conclusion: We demonstrated that tenocyte-like cells can be successfully differentiated from T-MSCs and BM-MSCs under TGF- β3 stimulation. This study demonstrated that T-MSCs and BM-MSCs in tenogenic stimulation with TGF-β3 have a similar tenogenic differentiation potential using RSM.

microRNA-265 Regulates Bone Marrow Mesenchymal Stem Cells Differentiation and Promotes Sepsis Lung Injury Repair via Transforming Growth Factor Beta 1

2022 ◽  
Vol 12 (2) ◽  
pp. 405-410
Author(s):  
Lian Tan ◽  
Xiongxiong Wang ◽  
Danqi Chen ◽  
Li Xu ◽  
Yudong Xu ◽  
...  
Keyword(s):  
Stem Cells ◽  
Bone Marrow ◽  
Mesenchymal Stem Cells ◽  
Lung Injury ◽  
Transforming Growth Factor Beta ◽  
Transforming Growth Factor ◽  
Alveolar Type ◽  
Injury Repair ◽  
Qrt Pcr ◽  
Marrow Mesenchymal Stem Cells

Our study investigates whether miR-265 regulates the differentiation of rat bone marrow mesenchymal stem cells (BMSCs) into alveolar type II epithelial cells (ATII) through TGF-β1 and promotes lung injury repair in rats with sepsis, thereby inhibiting sepsis progression. 25 patients with sepsis admitted to the Respiratory and Critical Care Medicine Department of the hospital and 17 normal controls were included. TGF-β1 level was measured by ELISA. miR-265 level was measured by qRT-PCR and AT II-related genes and proteins expression was analyzed by western blot and qRT-PCR. miR-265 expression was significantly higher in sepsis patients than normal group. Progenitor BMSCs were long and shuttle-shaped after 1 and 3 days of growth. Cultured MSCs had low expression of the negative antigen CD34 (4.32%) and high expression of the positive antigen CD44 (99.87%). TGF-β1 level was significantly increased with longer induction time, while miR-265 expression was significantly decreased in cell culture medium. miR-265 interference significantly decreased TGF-β1 expression. In conclusion, miR-265 inhibits BMSC differentiation to AT II via regulation of TGF-β1, thereby inhibiting sepsis progression.

scholarly journals Influence of mechanical and TGF-β3 stimulation on the tenogenic differentiation of tonsil-derived mesenchymal stem cells

2022 ◽  
Vol 23 (1) ◽  
Author(s):  
Jaeyeon Wee ◽  
Hyang Kim ◽  
Sang-Jin Shin ◽  
Taeyong Lee ◽  
Seung Yeol Lee
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Cell Differentiation ◽  
Mrna Expression ◽  
Mechanical Loading ◽  
Transforming Growth Factor Beta ◽  
Transforming Growth Factor ◽  
Mechanical Strain ◽  
Tenogenic Differentiation ◽  
Static Conditions

Abstract Background Organogenesis from tonsil-derived mesenchymal cells (TMSCs) has been reported, wherein tenogenic markers are expressed depending on the chemical stimulation during tenogenesis. However, there are insufficient studies on the mechanical strain stimulation for tenogenic cell differentiation of TMSCs, although these cells possess advantages as a cell source for generating tendinous tissue. The purpose of this study was to investigate the effects of mechanical strain and transforming growth factor-beta 3 (TGF-β3) on the tenogenic differentiation of TMSCs and evaluate the expression of tendon-related genes and extracellular matrix (ECM) components, such as collagen. Results mRNA expression of tenogenic genes was significantly higher when the mechanical strain was applied than under static conditions. Moreover, mRNA expression of tenogenic genes was significantly higher with TGF-β3 treatment than without. mRNA expression of osteogenic and chondrogenic genes was not significantly different among different mechanical strain intensities. In cells without TGF-β3 treatment, double-stranded DNA concentration decreased, while the amount of normalized collagen increased as the intensity of mechanical strain increased. Conclusions Mechanical strain and TGF-β3 have significant effects on TMSC differentiation into tenocytes. Mechanical strain stimulates the differentiation of TMSCs, particularly into tenocytes, and cell differentiation, rather than proliferation. However, a combination of these two did not have a synergistic effect on differentiation. In other words, mechanical loading did not stimulate the differentiation of TMSCs with TGF-β3 supplementation. The effect of mechanical loading with TGF-β3 treatment on TMSC differentiation can be manipulated according to the differentiation stage of TMSCs. Moreover, TMSCs have the potential to be used for cell banking, and compared to other mesenchymal stem cells, they can be procured from patients via less invasive procedures.

scholarly journals Effect of Lactoferrin on the Expression Profiles of Long Non-coding RNA during Osteogenic Differentiation of Bone Marrow Mesenchymal Stem Cells

2019 ◽  
Vol 20 (19) ◽  
pp. 4834 ◽  
Author(s):  
Yan Xu ◽  
Jing-Jing An ◽  
Dina Tabys ◽  
Yin-Dan Xie ◽  
Tian-Yu Zhao ◽  
...  
Keyword(s):  
Stem Cells ◽  
Bone Marrow ◽  
Mesenchymal Stem Cells ◽  
Osteogenic Differentiation ◽  
Transforming Growth Factor Beta ◽  
Transforming Growth Factor ◽  
Expression Profiles ◽  
Bioinformatic Analysis ◽  
Marrow Mesenchymal Stem Cells ◽  
The Impact

Lactoferrin (LF) has demonstrated stimulation of osteogenic differentiation of mesenchymal stem cells (MSCs). Long non-coding RNAs (lncRNAs) participate in regulating the osteogenic differentiation processes. However, the impact of LF on lncRNA expression in MSC osteogenic differentiation is poorly understood. Our aim was to investigate the effects of LF on lncRNAs expression profiles, during osteogenic differentiation of rat bone marrow mesenchymal stem cells (rBMSCs), by RNA sequencing. A total number of 1331 putative lncRNAs were identified in rBMSCs during osteogenic differentiation in the study. LF influenced the expression of 120 lncRNAs (differentially expressed lncRNAs [DELs], Fold change > 1.5 or < −1.5; p < 0.05) in rBMSCs on day 14 of osteogenic differentiation, consisted of 60 upregulated and 60 down-regulated. Furthermore, the potential functions of DELs were of prediction by searching their target cis- and trans-regulated protein-coding genes. The bioinformatic analysis of DELs target gene revealed that LF led to the disfunction of transforming growth factor beta stimulus (TGF-β) and positive regulation of I-κappa B kinase/NF-κappa B signaling pathway, which may relate to osteogenic differentiation of rBMSCs. Our work is the first profiling of lncRNA in osteogenic differentiation of rBMSCs induced by LF, and provides valuable insights into the potential mechanisms for LF promoting osteogenic activity.

Use of mesenchymal stem cells from adult bone marrow for injured tissue repair

2009 ◽  
Vol 150 (27) ◽  
pp. 1259-1265 ◽  
Author(s):  
Antal Salamon ◽  
Erzsébet Toldy
Keyword(s):  
Tissue Engineering ◽  
Stem Cells ◽  
Bone Marrow ◽  
Mesenchymal Stem Cells ◽  
Transforming Growth Factor Beta ◽  
Tissue Repair ◽  
Transforming Growth Factor ◽  
Injured Tissue ◽  
Growth Factor Beta ◽  
Adult Bone

A csontvelőből származó mesenchymalis őssejtek pluripotensek, s képesek porc, csont, valamint adiposus és ínsejtekké differenciálódni. Ezen mesenchymalis progenitor sejteket stromasejteknek vagy mesenchymalis őssejteknek nevezik. A csontvelőben két fő sejttípus van: haematopoeticus sejt és stromasejt. Mesenchymalis őssejtek kis beavatkozással nyerhetők a csontvelőből, majd sejtkultúrában szaporíthatóak. Differenciálódásuk bioaktív molekulákkal, specifikus növekedési faktorokkal segíthető elő. A transforming growth factor beta (TGF-β) család tagjai proteinek, közülük a bone morphogenetic proteinek (BMP) a legfontosabb faktorok, amelyek elősegítik a mesenchymalis őssejtek porc- és csontszövetté történő differenciálódását. Kevésbé ismert még ezen sejteknek a tenogenesisben való szerepe, de már vannak biztató adatok e téren is. A mesenchymalis őssejteknek és növekedési faktoroknak a sérült szövetekbe való juttatásra vivő vázanyagra (carrier, scaffold) van szükség. Mesenchymalis őssejtek használhatók fel génterápiára és a tissue engineering alkalmazására. A szerzők jelen munkájukban áttekintik a mesenchymalis őssejtek, biomolekulák és növekedési faktorok szövetpótlás céljából történő használatával foglalkozó kísérletes vizsgálatok eddigi eredményeit és ismertetik a klinikai alkalmazás lehetőségeit.

scholarly journals Mesenchymal stem cells targeting PI3K/AKT pathway in leukemic model

Tumor Biology ◽  
2019 ◽  
Vol 41 (4) ◽  
pp. 101042831984680 ◽  
Author(s):  
Esraa S A Ahmed ◽  
Neamat H Ahmed ◽  
Amina M Medhat ◽  
Ussama Z Said ◽  
Laila A Rashed ◽  
...  
Keyword(s):  
Stem Cells ◽  
Bone Marrow ◽  
Mesenchymal Stem Cells ◽  
Growth Factors ◽  
B Cell ◽  
Transforming Growth Factor Beta ◽  
Transforming Growth Factor ◽  
Cell Lymphoma ◽  
Hematological Parameters ◽  
B Cell Lymphoma

Mesenchymal stem cells have therapeutic properties that are related to their potentials for trans-differentiation, immunomodulation, anti-inflammatory, inhibitory effect on tumor proliferation, and induction of apoptosis. This study was performed to analyze the role of mesenchymal stem cells as an alternative for cellular signaling growth factors involved in the pathogenesis of leukemogenesis in rats. Treatment of rats with 7,12-dimethyl benz [a] anthracene induced leukemogenesis appeared as a significant decrease in hematological parameters with concomitant significant increase in bone marrow oxidative and inflammatory indices (transforming growth factor beta and interleukin-6) in comparison with normal groups. On the contrary, Western immunoblotting showed a significant increase in the signaling growth factors: PI3K, AKT, mTOR proteins and a significant decrease in PTEN in 7,12-dimethyl benz [a] anthracene–treated group. In addition, a significant increase in the transcript levels of B cell lymphoma-2 protein gene in the 7,12-dimethyl benz [a] anthracene group, while that of C-X-C motif chemokine receptor-4 and B cell lymphoma-2 protein associated x-protein were significantly downregulated compared to controls. Meanwhile, therapeutic mesenchymal stem cells treatment predict a significant improvement versus 7,12-dimethyl benz [a] anthracene group through the modulation of growth factors that confront bone marrow dysplasia. In the same direction treatment of 7,12-dimethyl benz [a] anthracene group with mesenchymal stem cells, it induced apoptosis and increased the homing efficacy to bone marrow. In conclusion, mesenchymal stem cells improve hematopoiesis and alleviate inflammation, and modulated PI3K/AKT signaling pathway contributed to experimental leukemogenesis.

scholarly journals Coordinated Regulation of Mesenchymal Stem Cell Migration by Various Chemotactic Stimuli

2020 ◽  
Vol 21 (22) ◽  
pp. 8561
Author(s):  
Donghyun Nam ◽  
Aran Park ◽  
Maria Jose Dubon ◽  
Jinyeong Yu ◽  
Wootak Kim ◽  
...  
Keyword(s):  
Stem Cells ◽  
Bone Marrow ◽  
Mesenchymal Stem Cells ◽  
Stem Cell ◽  
Mesenchymal Stem Cell ◽  
Transforming Growth Factor Beta ◽  
Transforming Growth Factor ◽  
Mouse Bone Marrow ◽  
Functional Interactions ◽  
Combination Treatments

Endogenous bone marrow-derived mesenchymal stem cells are mobilized to peripheral blood and injured tissues in response to changes in the expression of various growth factors and cytokines in the injured tissues, including substance P (SP), transforming growth factor-beta (TGF-β), and stromal cell-derived factor-1 (SDF-1). SP, TGF-β, and SDF-1 are all known to induce the migration of bone marrow-derived mesenchymal stem cells (BM-MSCs). However, it is not yet clear how these stimuli influence or interact with each other during BM-MSC mobilization. This study used mouse bone marrow-derived mesenchymal stem cell-like ST2 cells and human BM-MSCs to evaluate whether SP, TGF-β, and SDF-1 mutually regulate their respective effects on the mobilization of BM-MSCs. SP pretreatment of ST2 and BM-MSCs impaired their response to TGF-β while the introduction of SP receptor antagonist restored the mobilization of ST2 and BM-MSCs in response to TGF-β. TGF-β pretreatment did not affect the migration of ST2 and BM-MSCs in response to SP, but downregulated their migration in response to SDF-1. SP pretreatment modulated the activation of TGF-β noncanonical pathways in ST2 cells and BM-MSCs, but not canonical pathways. These results suggest that the migration of mesenchymal stem cells is regulated by complex functional interactions between SP, TGF-β, and SDF-1. Thus, understanding the complex functional interactions of these chemotactic stimuli would contribute to ensuring the development of safe and effective combination treatments for the mobilization of BM-MSCs.

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