Different Conditions for the Modification of Polycaprolactone Films with L-Arginine

Poly-ε-caprolactone (PCL) is a biodegradable polymer used in regenerative medicine. Mesenchymal stem cells (MSCs) play an important role in the regeneration of different tissues. The hydrophobicity and neutrality of a PCL surface reduce MSCs’ adhesion and proliferation. In this study, PCL films were treated with arginine to improve surface hydrophilicity. The influences of arginine concentration, temperature, and solvent on PCL surface properties were investigated. PCL films treated with a solution of arginine in isopropyl alcohol were found to have the maximum number of amino groups. The greatest number of cells, 2 h after seeding, adhered to such films. It was shown that amino groups affect the interaction of cells with a modified surface and the hydrolysis reaction after treatment with isopropyl alcohol promotes the formation of adhesive focal contacts. Hence, our results illustrate that functional groups on the PCL surface after arginine solution treatment regulate MSC adhesion and focal contact formation.

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Comparing the Chondrogenic Potential in vivo of Autogeneic Mesenchymal Stem Cells Derived from Different Tissues

Artificial Cells Blood Substitutes and Biotechnology ◽

10.3109/10731191003776769 ◽

2010 ◽

Vol 39 (1) ◽

pp. 31-38 ◽

Cited By ~ 35

Author(s):

Qiang Li ◽

Jicun Tang ◽

Riying Wang ◽

Chaoyong Bei ◽

Linwei Xin ◽

...

Keyword(s):

Stem Cells ◽

Mesenchymal Stem Cells ◽

Chondrogenic Potential ◽

Different Tissues

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Comparative adhesive and migratory properties of mesenchymal stem cells from different tissues

Biorheology ◽

10.3233/bir-180185 ◽

2019 ◽

Vol 56 (1) ◽

pp. 15-30 ◽

Cited By ~ 3

Author(s):

Asma Alanazi ◽

Hafsa Munir ◽

Mohammed Alassiri ◽

Lewis S.C. Ward ◽

Helen M. McGettrick ◽

...

Keyword(s):

Stem Cells ◽

Mesenchymal Stem Cells ◽

Different Tissues

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OPTIMIZATION of the Cultivation of Mesenchymal STEM CELLS for CLINICAL USE

Blood ◽

10.1182/blood.v118.21.4394.4394 ◽

2011 ◽

Vol 118 (21) ◽

pp. 4394-4394

Author(s):

Vanessa de Souza Valim ◽

Fernanda Oliveira ◽

Maria Aparecida Lima da Silva ◽

Bruna Amorin ◽

Lauro Moraes ◽

...

Keyword(s):

Stem Cells ◽

Mesenchymal Stem Cells ◽

Conflicts Of Interest ◽

Calf Serum ◽

Optimal Number ◽

Third Party ◽

Clinical Use ◽

Clinical Grade ◽

Plating Density ◽

Number Of Cells

Abstract Abstract 4394 Introduction and objectives: Studies with mesenchymal stem cells (MSCs) have shown its benefits in hematology, mainly for Graft-versus-host disease (Lancet 371:1579–86, 2008), with three unsettled matters: (1)MSCs expansion in medium supplemented with Fetal Calf Serum (FCS) and its risk of xenoreaction (Blood 89:776–9, 1997); (2)The optimal number of cells needed for therapy is not yet defined, but there is an empirical indication for 2×106células/Kg with the need to optimize expansion, number and time wise; and (3)the utilization of third party donors. This study was designed to determine the superiority or no-inferiority of the Platelet Lysates (PL) over FCS on the expansion of MSC, the optimal cell plating density and days between each pass, and to investigate if in our conditions total nucleated cells (TNC) obtained from the washouts of HSCT explants can expand to be used at clinical grade. Methods and Results: TNC were removed from the filters and bags used in the HSCT (Cytotherapy 11:403–13, 2009) and after the first passage were plated in different concentrations (2000/cm2, 3000/cm2, 4000/cm2, 5000/cm2, 6000/cm2 and 7000/cm2) with 10% FBS or 10% PL, and the number of days reach 80% of confluence was observe (Transfusion, 49:2680–5, 2009). Next, cultures with the same plating density were fed either with 10% PL or 10% FCS and were trypsinized after seven days and counted to analyze how much they have grown in that time period. And finally, cultures were allowed to growth up to Passage 3 (P3) to test the ability to obtain clinical grade number of cells. The proliferation of mesenchymal stem cells in the presence of PL and SFB was averaged 11.88 and 2.5 times, respectively, in a period of 7 days (p = 0.005). The highest concentration of plating cells using PL, took less time (6 days) to reach confluence as compared with the three lower (7.55 to 8.55 days) (p = 0.005), and at P3 with PL we obtained from 10×109 up to 10 × 1011 cells. Conclusion: This study suggests that the PL is the best choice as a supplement to expand MSC, and allow the proliferation of a sufficient number of MSC at P3 for clinical use obtained from the washouts of HSCT explants. Disclosures: No relevant conflicts of interest to declare.

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Characteristics of mesenchymal stem cells derived from different tissues of goat

Journal of Preventive Veterinary Medicine ◽

10.13041/jpvm.2019.43.3.107 ◽

2019 ◽

Vol 43 (3) ◽

pp. 107-114

Author(s):

Na-Yeon Gu ◽

◽

Da-Un Jeong ◽

Jeong Su Byeon ◽

Jae-Young Song ◽

...

Keyword(s):

Stem Cells ◽

Mesenchymal Stem Cells ◽

Different Tissues

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Mesenchymal Stem Cells from Different Tissues: Immune Status and Activity

Journal of Immunology and Infectious Diseases ◽

10.15744/2394-6512.3.102 ◽

2016 ◽

Vol 3 (1) ◽

Author(s):

Trivanović D ◽

Krstić J ◽

Mojsilović S ◽

Djordjević IO ◽

Ilić V ◽

...

Keyword(s):

Stem Cells ◽

Mesenchymal Stem Cells ◽

Immune Status ◽

Different Tissues

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Comparison of biological features of wild European rabbit mesenchymal stem cells derived from different tissues.

10.1101/2021.12.13.472420 ◽

2021 ◽

Author(s):

Marta Díaz-de Frutos ◽

Alexandra Calle ◽

María Zamora-Ceballos ◽

Juan Bárcena ◽

Esther Blanco ◽

...

Keyword(s):

Stem Cells ◽

Mesenchymal Stem Cells ◽

Adipose Tissue ◽

European Rabbit ◽

Isolation And Characterization ◽

Expression Characteristic ◽

Subcutaneous Adipose ◽

Different Tissues

Although the European rabbit is an "endangered" species and a notorious biological model, the analysis and comparative characterization of new tissue sources of rabbit mesenchymal stem cells (rMSCs) has not been well studied. Here we report for the first time the isolation and characterization of rMSCs derived from an animal belonging to a natural rabbit population within the species native region. New rMSC lines were isolated from different tissues: oral mucosa (rOM-MSC), dermal skin (rDS-MSC), subcutaneous adipose tissue (rSCA-MSC), ovarian adipose tissue (rOA-MSC), oviduct (rO-MSC), and mammary gland (rMGMSC). The six rMSC lines showed plastic adhesion with fibroblast-like morphology and were all shown to be positive for CD44 and CD29 expression (characteristic markers of MSCs), and negative for CD34 or CD45 expression. In terms of pluripotency features, all rMSC lines expressed NANOG, OCT4, and SOX2. Furthermore, all rMSC lines cultured under osteogenic, chondrogenic, and adipogenic conditions showed differentiation capacity. In conclusion, this study describes the isolation and characterization of new rabbit cell lines from different tissue origins, with a clear mesenchymal pattern. We show that rMSC do not exhibit differences in terms of morphological features, expression of the cell surface, and intracellular markers of pluripotency and in vitro differentiation capacities, attributable to their tissue of origin.

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

BioMed Research International ◽

10.1155/2019/6376271 ◽

2019 ◽

Vol 2019 ◽

pp. 1-13 ◽

Cited By ~ 17

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.

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283 COMPARATIVE CHARACTERIZATIONS OF PORCINE MESENCHYMAL STEM CELLS AND SKIN-DERIVED CELLS

Reproduction Fertility and Development ◽

10.1071/rdv21n1ab283 ◽

2009 ◽

Vol 21 (1) ◽

pp. 238

Author(s):

S. A. Ock ◽

B. G. Jeon ◽

D. O. Kwack ◽

G. J. Rho

Keyword(s):

Stem Cells ◽

Mesenchymal Stem Cells ◽

Mrna Expression ◽

Solution Treatment ◽

Cell Types ◽

Skin Substitute ◽

Fetal Skin ◽

Santa Cruz ◽

Skin Cells ◽

Polyclonal Igg

Mesenchymal stem cells (MSC) have high potential in regenerative medicine such as skin substitute for a burn healing. The present study compared the characterizations of porcine MSCs (pMSC) derived from bone marrow to the same porcine adult ear and different porcine fetal skin derived cells on morphology of cell growth, alkaline phosphatase (AP) activity, cell surface (CD) markers (CD 29, 45, 90 and 105) and cell cycle by flow cytometer and protein and mRNA expression of Oct3/4, Nanog and Sox2 by immunocytochemistry and RT-PCR (Carlin et al. 2006 Reprod. Biol. Endocrinol. 4, 8) in 1–3 passage. The pMSC were isolated with Ficoll–Paque Plus (Amersham Science, USA) and skin-derived cells were separated with trypsin-EDTA solution treatment of tissue. Basal medium used for culture of pMSC and skin-derived cells was advanced-DMEM supplemented with 10% FBS and 1% penicillin–streptomycin (10 000 IU and 10 000 μg mL–1, Gibco) and culture medium was exchanged every 3 days. Skin-derived cells were observed similar patterns of colony formation and AP expression by AP chromogen kit (BCIP/NBT) (Abcam Inc., MA, USA) to pMSC on plating culture. pMSC and skin-derived cells were found to have a stronger reaction of CD 45– (St. Louis, MO, USA), CD 29+ (BD Bioscience, US), and CD 90+ (BD Bioscience, US). However, CD 105+ (abD Serotec, UK) weakly expressed in pMSC and skin-derived cells were almost negative. The population of cells at S-phase of the cycle in pMSC was significantly (P < 0.05) greater than those in adult ear and fetal-derived skin cells (30.4 ± 5.3 v. 19.1 ± 3.6 and 21.3 ± 1.7, respectively; mean ± SD). Protein of Oct3/4 (goat polyclonal IgG, Santa Cruz, CA, USA), Nanog (goat polyclonal IgG, Santa Cruz, CA, USA) and Sox2 (rabbit polyclonal IgG, Santa Cruz, CA, USA) were observed at both nucleus and cytoplasm in all cell types. However, the pattern of mRNA expression was different among cell types. Nanog and Sox2 expression was the greatest in pMSC and fetal skin cells, respectively. Oct 4 expression should read extremely low in pMSC compared with skin-derived cells, contrary to expectation. Taken together, pMSC and skin-derived cells revealed similar characterization, indicating that pMSC can expect a role of skin substitute. This work was supported by grant from Biogreen21 (20070301034040), Ministry of Agriculture and Forestry, Republic of Korea.

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Study on Pipetting Motion Optimization of Automatic Spheroid Culture System for Spheroid Formation

Journal of Robotics and Mechatronics ◽

10.20965/jrm.2021.p0078 ◽

2021 ◽

Vol 33 (1) ◽

pp. 78-87

Author(s):

Takeshi Shimoto ◽

Chihiro Teshima ◽

Toshiki Watanabe ◽

Xiu-Ying Zhang ◽

Atsushi Ishikawa ◽

...

Keyword(s):

Stem Cells ◽

Mesenchymal Stem Cells ◽

Culture System ◽

Three Dimensional ◽

Spheroid Formation ◽

Spheroid Culture ◽

Motion Optimization ◽

Passage Number ◽

Dimensional Cell ◽

Number Of Cells

This research group has established a technology for producing a three-dimensional cell constructed using only the cell itself. This technology uses a property in which the spheroids fuse with each other. We developed a system that automates the spheroid production process to obtain reproducible spheroids and suppress variation factors that occur from human operation. However, it has become clear that the dispersion occurs in the diameter depending on the number of cells of the spheroid even if the cells are handled in the same manner. The purpose of this research is to examine an appropriate pipetting motion in accordance with the number of cells of the spheroid to be produced. Rabbit mesenchymal stem cells (rMSCs) are used as the objects. The number of cells was set to 2×104, 3×104, and 4×104 cells/well, and the passage number as 7. The appearance of spheroids cultured using the motion programmed in accordance with each number of cells was observed every 24 hours for 5 days after seeding. The results of the analysis indicate that the optimum motion in each number of cells has been successfully specified, and reproducible spheroids have been successfully produced.

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