Immunophenotypic characterization and tenogenic differentiation of mesenchymal stromal cells isolated from equine umbilical cord blood

Mohanty, Nihar Nalini; Gulati, Baldev R.; Kumar, Rajesh; Gera, Sandeep; Kumar, Pawan; Somasundaram, Rajesh; Kumar, Sandeep

doi:10.1007/s11626-013-9729-7

Cited by 32 publications

(23 citation statements)

References 45 publications

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“…3c. The doubling time of cells in SVF solution cultured on TCPS dishes by the conventional culture method appeared to be approximately 2.0 days, which is similar to the doubling time reported by several investigators141727. The doubling time of the migrated cells from NY mesh filters having a pore size of 11 μm showed similar doubling time, whereas the doubling time of the migrated cells from the filter having pore size >20 μm that were cultured on PS dishes was longer than the doubling time of the cells in SVF solution cultured on TCPS dishes ( p < 0.05).…”

Section: Resultssupporting

confidence: 87%

Purification and differentiation of human adipose-derived stem cells by membrane filtration and membrane migration methods

Lin

Heish

Liu

et al. 2017

Sci Rep

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Human adipose-derived stem cells (hADSCs) are easily isolated from fat tissue without ethical concerns, but differ in purity, pluripotency, differentiation ability, and stem cell marker expression, depending on the isolation method. We isolated hADSCs from a primary fat tissue solution using: (1) conventional culture, (2) a membrane filtration method, (3) a membrane migration method where the primary cell solution was permeated through membranes, adhered hADSCs were cultured, and hADSCs migrated out from the membranes. Expression of mesenchymal stem cell markers and pluripotency genes, and osteogenic differentiation were compared for hADSCs isolated by different methods using nylon mesh filter membranes with pore sizes ranging from 11 to 80 μm. hADSCs isolated by the membrane migration method had the highest MSC surface marker expression and efficient differentiation into osteoblasts. Osteogenic differentiation ability of hADSCs and MSC surface marker expression were correlated, but osteogenic differentiation ability and pluripotent gene expression were not.

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Section: Resultssupporting

confidence: 87%

Purification and differentiation of human adipose-derived stem cells by membrane filtration and membrane migration methods

Lin

Heish

Liu

et al. 2017

Sci Rep

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“…BMP‐7, ‐12, ‐13, and ‐14 have been implicated in the neoformation and repair of tendons, and BMP‐12 has specifically been shown to promote tendon differentiation and formation in vitro and in vivo . Multiple studies have shown that BMP‐12 alone is sufficient to promote tenogenic differentiation of MSCs in vitro , as observed through increased expression of tendon markers, including tenomodulin, decorin, and scleraxis . Interestingly, for rat MSCs on collagen sponges, increases in the expression of scleraxis and tenomodulin were observed over 14 days, after only 12 h of exposure to BMP‐12 on day 1.…”

Section: Soluble Signaling Cuesmentioning

confidence: 99%

“…[116][117][118] Multiple studies have shown that BMP-12 alone is sufficient to promote tenogenic differentiation of MSCs in vitro, as observed through increased expression of tendon markers, including tenomodulin, decorin, and scleraxis. 118,119 118 Equine BMSCs BMP-12 Increased tenomodulin and decorin expression by MSCs in BMP-12 on day 20 Park et al 123 Rat ADSCs GDF-5 Increased cell number at 100 ng/mL at days 3-12; increased expression of scleraxis and tenomodulin in 100 ng/mL, increased tenascin-C expression in 1000 ng/mL Zhang et al 130 Human Human BMSCs FGF versus EGF + TGF-␤1…”

Section: Matrix Mechanical Propertiesmentioning

confidence: 99%

Designing the stem cell microenvironment for guided connective tissue regeneration

Bogdanowicz

2017

Annals of the New York Academy of Sciences

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Adult mesenchymal stem cells (MSCs) are an attractive cell source for regenerative medicine because of their ability to self-renew and their capacity for multilineage differentiation and tissue regeneration. For connective tissues, such as ligaments or tendons, MSCs are vital to the modulation of the inflammatory response following acute injury while also interacting with resident fibroblasts to promote cell proliferation and matrix synthesis. To date, MSC injection for connective tissue repair has yielded mixed results in vivo, likely due to a lack of appropriate environmental cues to effectively control MSC response and promote tissue healing instead of scar formation. In healthy tissues, stem cells reside within a complex microenvironment comprising cellular, structural, and signaling cues that collectively maintain stemness and modulate tissue homeostasis. Changes to the microenvironment following injury regulate stem cell differentiation, trophic signaling, and tissue healing. Here, we focus on models of the stem cell microenvironment that are used to elucidate the mechanisms of stem cell regulation and inspire functional approaches to tissue regeneration. Recent studies in this frontier area are highlighted, focusing on how microenvironmental cues modulate MSC response following connective tissue injury and, more importantly, how this unique cell environment can be programmed for stem cell-guided tissue regeneration.

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“…Equine MSCs are commonly harvested from bone marrow or adipose tissue and are expanded in vitro before use in autologous transplants [10–13]. The requirement to use surgical procedures to harvest cells from those locations has driven the search for other—less invasive—sources including whole blood, umbilical cord blood, or Wharton jelly [14–17]. In that regard, the endometrium represents an attractive alternative source of MSCs in the horse.…”

Section: Introductionmentioning

confidence: 99%

Isolation and characterization of equine endometrial mesenchymal stromal cells

et al. 2017

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BackgroundEquine mesenchymal stromal/stem cells (MSCs) are most commonly harvested from bone marrow (BM) or adipose tissue, requiring the use of surgical procedures. By contrast, the uterus can be accessed nonsurgically, and may provide a more readily available cell source. While human endometrium is known to harbor mesenchymal precursor cells, MSCs have not been identified in equine endometrium. This study reports the isolation, culture, and characterization of MSCs from equine endometrium.MethodsThe presence of MSC and pericyte markers in endometrial sections was determined using immunohistochemistry. Stromal cells were harvested and cultured after separation of epithelial cells from endometrial fragments using Mucin-1-bound beads. For comparison, MSCs were also harvested from BM. The expression of surface markers in endometrial and BM-derived MSCs was characterized using flow cytometry and quantitative polymerase chain reaction. MSCs were differentiated in vitro into adipogenic, chondrogenic, osteogenic, and smooth muscle lineages.ResultsTypical markers of MSCs (CD29, CD44, CD90, and CD105) and pericytes (NG2 and CD146) were localized in the equine endometrium. Both endometrial and BM MSCs grew clonally and robustly expressed MSC and pericyte markers in culture while showing greatly reduced or negligible expression of hematopoietic markers (CD45, CD34) and MHC-II. Additionally, both endometrial and BM MSCs differentiated into adipogenic, osteogenic, and chondrogenic lineages in vitro, and endometrial MSCs had a distinct ability to undergo smooth muscle differentiation.ConclusionsWe have demonstrated for the first time the presence of cells in equine endometrium that fulfill the definition of MSCs. The equine endometrium may provide an alternative, easily accessible source of MSCs, not only for therapeutic regeneration of the uterus, but also for other tissues where MSCs from other sources are currently being used therapeutically.

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Immunophenotypic characterization and tenogenic differentiation of mesenchymal stromal cells isolated from equine umbilical cord blood

Cited by 32 publications

References 45 publications

Purification and differentiation of human adipose-derived stem cells by membrane filtration and membrane migration methods

Purification and differentiation of human adipose-derived stem cells by membrane filtration and membrane migration methods

Designing the stem cell microenvironment for guided connective tissue regeneration

Isolation and characterization of equine endometrial mesenchymal stromal cells

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