Comparative studies on proliferation, molecular markers and differentiation potential of mesenchymal stem cells from various tissues (adipose, bone marrow, ear skin, abdominal skin, and lung) and maintenance of multipotency during serial passages in miniature pig

Lee, Ah‐Young; Lee, Jienny; Kim, Chan-Lan; Lee, Keum Sil; Lee, SoHyun; Gu, Na-Yeon; Kim, Jeong‐Min; Lee, Byeong Chun; Koo, Ok Jae; Song, Jae‐Young; Cha, Sang‐Ho

doi:10.1016/j.rvsc.2015.03.010

Cited by 43 publications

(35 citation statements)

References 32 publications

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“…Molecular mechanisms involved in the regulation of MSC properties, in particular stemness features, are not completely understood . Nanog and Oct4 genes have been reported to be expressed by MSCs from different species and to play a role in preserving their self‐renewal capacity and undifferentiated state . Also Dnmt1 and p21 have been suggested to influence these properties in human MSCs, in particular they have been proposed to exert opposite functions one to the other: while increased expression of the former prevents senescence and sustains proliferation and differentiation capacity, higher levels of the latter reduce stemness markers expression and proliferative potential .…”

Section: Discussionmentioning

confidence: 99%

Murine Rankl−/− Mesenchymal Stromal Cells Display an Osteogenic Differentiation Defect Improved by a RANKL-Expressing Lentiviral Vector

Schena¹,

Menale

Caci

et al. 2017

Stem Cells

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Autosomal recessive osteopetrosis (ARO) is a severe bone disease characterized by increased bone density due to impairment in osteoclast resorptive function or differentiation. Hematopoietic stem cell transplantation is the only available treatment; however, this therapy is not effective in RANKL-dependent ARO, since in bone this gene is mainly expressed by cells of mesenchymal origin. Of note, whether lack of RANKL production might cause a defect also in the bone marrow (BM) stromal compartment, possibly contributing to the pathology, is unknown. To verify this possibility, we generated and characterized BM mesenchymal stromal cell (BM-MSC) lines from wild type and Rankl mice, and found that Rankl BM-MSCs displayed reduced clonogenicity and osteogenic capacity. The differentiation defect was significantly improved by lentiviral transduction of Rankl BM-MSCs with a vector stably expressing human soluble RANKL (hsRANKL). Expression of Rankl receptor, Rank, on the cytoplasmic membrane of BM-MSCs pointed to the existence of an autocrine loop possibly activated by the secreted cytokine. Based on the close resemblance of RANKL-defective osteopetrosis in humans and mice, we expect that our results are also relevant for RANKL-dependent ARO patients. Data obtained in vitro after transduction with a lentiviral vector expressing hsRANKL would suggest that restoration of RANKL production might not only rescue the defective osteoclastogenesis of this ARO form, but also improve a less obvious defect in the osteoblast lineage, thus possibly achieving higher benefit for the patients, when the approach is translated to clinics. Stem Cells 2017;35:1365-1377.

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Section: Discussionmentioning

confidence: 99%

Murine Rankl−/− Mesenchymal Stromal Cells Display an Osteogenic Differentiation Defect Improved by a RANKL-Expressing Lentiviral Vector

Schena¹,

Menale

Caci

et al. 2017

Stem Cells

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“…One million passage 3 ASCs were suspended in 100 μL PBS containing 10 μg/ml fluorescein isothiocyanate-conjugated primary antibodies specific to mesenchymal stromal cells (MSCs) (CD29, CD44, CD90, and CD105) and hematopoietic cells (CD31 and CD45) (n = 3). The following expression markers reactive with the porcine antigen isoforms were used: Alexa Fluor 647 Mouse Anti-Pig CD29 (BD Bioscience, New Jersey, USA), Anti-CD44 antibody [IM7] (Abcam plc, Cambridge, UK), APC Mouse Anti-Human CD90 (BD-Biosciences), Anti-CD105 antibody [MEM-229] (Abcam plc), PE Mouse Anti-Rat CD31 (BD Biosciences), and Monoclonal Antibody to CD45/LCA (CD45R)-PE (Acris Antibodies, Inc. CA, USA) [24] , [25] , [26] , [27] , [28] . Cell fluorescence was evaluated with a Gallios flow cytometer (Beckman Coulter, Tokyo, Japan) and the data were analyzed using Karuza for Gallios software (Beckman Coulter).…”

Section: Methodsmentioning

confidence: 99%

Autologous adipose-derived stem cell sheets enhance the strength of intestinal anastomosis

Maruya¹,

Kanai²,

Kobayashi³

et al. 2017

Regenerative Therapy

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ObjectiveAdipose-derived stem cells (ASCs) are capable of multiple differentiation pathways, imparting immunomodulatory effects, and secreting factors that are important for wound healing. These characteristics can be exploited to decrease the incidence of anastomotic leakage.MethodsIn order to delay local wound healing at the anastomotic site, we induced ischemia in a portion of porcine small intestine by ligating vessels. Then, we injected mitomycin C into the serosa of the small intestine above the ligated vessels. Anastomotic sites were created by 2 cm incisions made in the opposite mesenteric area. ASCs were isolated from the porcine subcutaneous fat tissues and expanded under culture conditions. ASCs were trypsinized and seeded on temperature-responsive dishes and cultured to form confluent sheets. Three ASC sheets were transplanted onto the serous membrane after suturing. The extent of anastomotic wound healing was evaluated by bursting pressure, hydroxyproline content, and mRNA expression of collagen-1 alpha1 and collagen-3 alpha1.ResultsWe found that transplantation of ASC sheets increased anastomotic site bursting pressure. Additionally, transplantation of ASC sheets increased the hydroxyproline content of the anastomoses. Furthermore, transplantation of ASC sheets increased mRNA expression of collagen-1 alpha1 and collagen-3 alpha1.ConclusionsOur findings showed that transplantation of autologous ASC sheets enhanced collagen synthesis and anastomotic strength. Further studies are necessary to identify substances that, in combination with ASC sheets, might enhance collagen synthesis and healing in sites of anastomosis.

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“…7,28,29 These include comparable morphology, proliferative capacity, alkaline phosphatase activity, cell surface marker expression, metabolic pathways, biologic functions, and transcription factors. 10,23,30 pASCs have often been compared side-by-side with porcine stem cells from these other tissue sources, revealing characteristics and comparable multilineage differentiation abilities as well. 9,23,28 While knowledge of the differentiation abilities of pASCs is currently limited to a few reports, porcine bone marrow-derived stem cells (pBMSCs) have been differentiated into myocytes, 31 endothelial cells, 32 and epithelial cells.…”

Section: Multilineage Differentiation Abilitiesmentioning

confidence: 99%

“…Typical cell seeding densities range from 5000 to 7000 cells/cm 2 . 5,9,10 Dulbecco's Modified Eagle Medium (DMEM) mixed 1:1 with Ham's F-12 Nutrient Mixture and supplemented with 10% Fetal Bovine Serum has been reported as ideal for the culture of pASCs. 5,10 After 48/72hrs in culture the non-adherent haematopoietic cells are removed.…”

mentioning

confidence: 99%

Properties of porcine adipose-derived stem cells and their applications in preclinical models

Arrizabalaga

Nollert

2017

Adipocyte

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Adipose-derived stem cells represent a reliable adult stem cell source thanks to their abundance, straightforward isolation, and broad differentiation abilities. Consequently, human adipose-derived stem cells (hASCs) have been used in vitro for several innovative cellular therapy and regenerative medicine applications. However, the translation of a novel technology from the laboratory to the clinic requires first to evaluate its safety, feasibility, and potential efficacy through preclinical studies in animals. The anatomy and physiology of pigs and humans are very similar, establishing pigs as an attractive and popular large animal model for preclinical studies. Knowledge of the properties of porcine adipose-derived stem cells (pASCs) used in preclinical studies is critical for their success. While hASCs have been extensively studied this past decade, only a handful of reports relate to pASCs. The aim of this concise review is to summarize the current findings about the isolation of pASCs, their culture, proliferation, and immunophenotype. The differentiation abilities of pASCs and their applications in porcine preclinical models will also be reported.

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Comparative studies on proliferation, molecular markers and differentiation potential of mesenchymal stem cells from various tissues (adipose, bone marrow, ear skin, abdominal skin, and lung) and maintenance of multipotency during serial passages in miniature pig

Cited by 43 publications

References 32 publications

Murine Rankl−/− Mesenchymal Stromal Cells Display an Osteogenic Differentiation Defect Improved by a RANKL-Expressing Lentiviral Vector

Murine Rankl−/− Mesenchymal Stromal Cells Display an Osteogenic Differentiation Defect Improved by a RANKL-Expressing Lentiviral Vector

Autologous adipose-derived stem cell sheets enhance the strength of intestinal anastomosis

Properties of porcine adipose-derived stem cells and their applications in preclinical models

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