Isolation, characterization and osteogenic differentiation of adipose-derived stem cells: from small to large animal models

Arrigoni, E.; Lopa, Silvia; Girolamo, Laura de; Stanco, D.; Brini, A.T.

doi:10.1007/s00441-009-0883-x

Cited by 108 publications

(108 citation statements)

References 41 publications

(57 reference statements)

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“…In this study, we isolated human MSCs from adipose tissue and bone marrow of the same donors to compare the morphology, proliferation ability, surface antigens and cardiomyogenic differentiation capacity of the two populations. Consistent with other studies (17,18), no difference in morphology was observed between ASCs and BMSCs. Prior to treatment with 5-azacytidine, both ASCs and BMSCs stained positively for CD29, CD44 and CD105, although negatively for CD34 and CD45; however, the difference in surface marker expression appears to be with CD49d and CD106.…”

Section: Discussionsupporting

confidence: 92%

Differentiation potential of human mesenchymal stem cells derived from adipose tissue and bone marrow to sinus node-like cells

Yang

Song

et al. 2011

Mol Med Report

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Abstract. Adult mesenchymal stem cells (MSCs) hold great promise for the repair of heart defects. Both bone marrowderived mesenchymal stem cells (BMSCs) and adipose tissue-derived stem cells (ASCs) are multipotent and may be induced by 5-azacytidine to differentiate into cardiomyocytes. However, the differentiation potential of human MSCs into sinus node-like cells has not been studied extensively. The aim of this study was to analyze the differences in proliferation and phenotype of ASCs and BMSCs from the same donors and to evaluate their capacity to differentiate into sinus node-like cells in vitro. Five passaged cells from bone marrow and adipose tissue were treated with 10 µM 5-azacytidine for 48 h and further cultured in complete medium for 4 weeks. A comparative study of cultured ASCs and BMSCs was carried out, and the morphological parameters, proliferative capacity, expression of surface markers and differentiation potential to sinus node-like cells were characterized. No morphologic differences were observed between ASCs and BMSCs. Flow cytometric analysis revealed that ASCs and BMSCs both expressed CD29, CD44, CD90 and CD105 and did not express CD34 and CD14, while CD49d, CD106 and CD34 were differentially expressed. Growth curves and doubling time determined with the Cell Counting Kit-8 (CCK-8) demonstrated that ASCs had a stronger proliferative ability than BMSCs. Histological immuofluorescence staining suggested that ASCs and BMSCs were capable of differentiating into sinus node-like cells and that the positive expression ratios of cTNI were higher in ASCs compared to BMSCs at 4 weeks. Expression of the HCN2 and HCN4 genes was detected by reverse transcriptase polymerase chain reaction, and the results revealed that the expression of the HCN genes appeared earlier in ASC-derived sinus node-like cells. ASCs expressed HCN2 and HCN4 shortly after induction with 5-azacytidine for 2 weeks, although BMSCs expressed these genes after 4 weeks. The expression levels of HCN2 and HCN4 mRNA in ASC-derived cells were higher compared to those of BMSCs at 4 weeks. In conclusion, ASCs may be a better candidate as a novel source of cell therapy in sinus bradycardia disorders than BMSCs.

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Section: Discussionsupporting

confidence: 92%

Differentiation potential of human mesenchymal stem cells derived from adipose tissue and bone marrow to sinus node-like cells

Yang

Song

et al. 2011

Mol Med Report

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“…On the ultrastructural level, they display normal morphology of the protheosynthetically active cells. Similar observations from experiments with somatic stem cells originated from various tissues were recorded by our group and other authors, not only in human but also in other species [7], [8], [10]- [12].…”

Section: Discussionsupporting

confidence: 76%

Do Dermis-Derived Stem Cells Have the Same Characteristics as Bone Marrow- and Adipose Tissue-Derived Stem Cells?

Boháč¹,

Polák²,

Fedels³

et al. 2013

IJBBB

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Abstract-In this study we performed comprehensive biological and morphological analysis of dermis-derived stem cells and compared them with bone marrow-and adipose tissue-derived stem cells. According to obtained results we can emphasize that analyzed somatic stem cells shared morphological features, expression of surface antigens as well as growth kinetics, but differ in chondrogenic potential. Skin-derived stem cells have an inferior potential for chondrogenic differentiation. So this fact decreases their potential for cartilage tissue engineering.Index Terms-Biological characteristics, chondrogenic differentiation, morphology, somatic stem cells. I. INTRODUCTIONDamaged or lost articular cartilage as a consequence of inborn anomaly, pathological process or trauma leads to progressive debilitation, which has major impact on the life quality of the affected individuals in all age groups. It is difficult to treat these patients, because mature articular cartilage has a limited self-repairing potential [1]. Small defects are regenerated by migration of chondrocytes, while full-thickness damages are healed by formation of inferior fibrocartilage [2]. However, in many cases osteoarthritis is developed and surgical intervention is often the only option. Unfortunately, current treatment techniques for cartilage reparation are insufficient and it is not possible to obtain expected results.Recently, somatic stem cells / mesenchymal stem cellsMSCs represent big hope in this respect. Stem cells are found in all tissues of higher multicellular organisms during all stages of ontogenesis. They are primitive undifferentiated cells with the unique ability of long term self-renewing. Another important feature of MSCs is their plasticity -the potential for terminal differentiation into other cell types. Therefore, they play an important role in the embryonic Manuscript received May 10, 2013; revised July 16, 2013 development, replacing worn out cells and regeneration of damaged tissues [3].MSCs are adherent and have a fibroblast-like morphology. They are also able to produce colony forming units-fibroblast (CFU-F) when cultured in vitro. They are positive for a variety of surface markers, including CD29, CD44, CD56, CD73, CD90, CD105, CD166, CD271, STRO-1 and Sca-1. On the contrary, they are negative for CD31, CD34, CD45 and HLA-DR [4]. More recently, it was shown that MSCs express markers which are typical for embryonic stem cells, including OCT4, Nanog and Sox2. These findings provide clarification of their undifferentiated state [5].Bone marrow was the first source of MSCs but in respect to clinical application it has several disadvantages, including invasive and painful sampling procedure and the decline in MSCs number and differentiation potential with increasing age of donor [6]. For that reason, alternative sources of MSCs are subject to intensive investigation. In that context, attention turns to adipose tissue which is abundant in human body and represents reliable and relatively safe source of MSCs (one gram of adipose...

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“…The isolation and culture of rASCs were performed, as previously described. 18,19 After rinsing in 0.25% chloromycetin and phosphate-buffered saline (PBS), the fresh adipose tissues were cut into small pieces and then treated with 0.10% collagenase I (Worthington Biochemical Corp.) under shaking at 37°C for 60 min. After digestion, the collagenase I was neutralized with low-glucose Dulbecco's modified Eagle's medium (LG-DMEM; Gibco) supplemented with 10% fetal bovine serum (FBS; Gibco), and the suspension was filtered through a 200-mm nylon mesh to remove the undigested tissue and then centrifuged at 1200 g for 10 min.…”

Section: Isolation and Culture Of Rascs In Vitromentioning

confidence: 99%

Epithelial-Differentiated Adipose-Derived Stem Cells Seeded Bladder Acellular Matrix Grafts for Urethral Reconstruction: An Animal Model

Xie

et al. 2014

Tissue Engineering Part A

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The limited amount of available epithelial tissue is considered a main cause of the high rate of urethral reconstruction failures. The aim of this study was to investigate whether epithelial-differentiated rabbit adiposederived stem cells (Epith-rASCs) could play a role of epithelium in vivo functionally and be a potential substitute of urothelium. Substitution urethroplasty was performed to repair an anterior urethral defect in male New Zealand rabbits using Epith-rASCs seeded bladder acellular matrix grafts (BAMGs) after 5-bromo-2¢-deoxyuridine (BrdU) labeling, based on the in vitro epithelial induction system we previously described. Urethroplasty with cell-free BAMGs and with undifferentiated rASCs (Und-rASCs) seeded BAMGs were performed as controls. After surgery, a notable amelioration of graft contracture and recovery of urethral continuity were observed in the Epith-rASCs/BAMG group by retrograde urethrograms and macroscopic inspection. Immunofluorescence revealed that the BrdU-labeled Epith-rASCs/Und-rASCs colocalized with cytokeratin 13 or myosin. Consistent with the results of western blotting, at early postimplantation stage, the continuous epithelial layer with local multilayered structure was observed in the Epith-rASCs/BAMG group, whereas no significant growth and local monolayer growth profile of epithelial cells were observed in the BAMG and Und-rASCs/BAMG group, respectively. The results showed that Epith-rASCs could serve as a potential substitute of urothelium for urethral tissue engineering and be available to prevent lumen contracture and subsequent complications including recurrent stricture.

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Isolation, characterization and osteogenic differentiation of adipose-derived stem cells: from small to large animal models

Cited by 108 publications

References 41 publications

Differentiation potential of human mesenchymal stem cells derived from adipose tissue and bone marrow to sinus node-like cells

Differentiation potential of human mesenchymal stem cells derived from adipose tissue and bone marrow to sinus node-like cells

Do Dermis-Derived Stem Cells Have the Same Characteristics as Bone Marrow- and Adipose Tissue-Derived Stem Cells?

Epithelial-Differentiated Adipose-Derived Stem Cells Seeded Bladder Acellular Matrix Grafts for Urethral Reconstruction: An Animal Model

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