Gelatin/PLLA Sponge‐Like Scaffolds Allow Proliferation and Osteogenic Differentiation of Human Mesenchymal Stromal Cells

Mattii, Letizia; Battolla, Barbara; D’Alessandro, Delfo; Trombi, Luisa; Pacini, Stefania; Cascone, Maria Grazia; Lazzeri, Luigi; Bernardini, Nunzia; Dolfi, Amelio; Galimberti, Sara; Petrini, Mario

doi:10.1002/mabi.200700331

Cited by 25 publications

(26 citation statements)

References 22 publications

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“…16 HMSCs from both sources were preliminarily characterized in our laboratories and showed no appreciable differences in terms of osteogenic potential. According to the planned seeding schedule (Table 1), aliquots of hMSCs were defrosted from liquid nitrogen storage and plated in tissue culture polystyrene (TCPS) flasks at a cell density of 1000 cells/cm 2 to reach the desired cell number in identical conditions (70% confluence, passage 2) at the scheduled times.…”

Section: Expansion and Culture Of Hmscsmentioning

confidence: 99%

“…Samples were processed for histology, as previously reported. 16 The sections, 8 mm thick, were stained with a specific kit for Alcian Blue (Bio-Optica), according to the manufacturer's recommendations.…”

Section: Osteogenic Differentiation Of the Niches: Qualitative And Sementioning

confidence: 99%

“…Such scaffolds were selected as they resulted to be highly suitable for both hMSC and osteoblast colonization on the basis of previous studies. [16][17][18][19] Coexistence of multistage osteogenic cells in the niches could be simply obtained by periodic seeding of undifferentiated hMSCs on hMSC/scaffold constructs, the latter being cultured in the osteogenic CM. In this way, owing to the time elapsed between each cell inoculation (i.e., 5 days), we artificially created simple cell-dynamic systems in which osteogenic cell gradients evolving with time have been generated.…”

mentioning

confidence: 99%

“…Samples (n = 2) at two time points, that is, (t 0 + 1 day) and (t f ), were processed for scanning electron microscopy (SEM) analysis as described earlier. 16 The dried samples were cross-sectioned, sputter-coated with gold (Sputter coater Emitech K550; Quorum Technologies Ltd.), and examined on a Quanta scanning electron microscope (FEI).…”

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confidence: 99%

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Growing Bone Tissue-Engineered Niches with Graded Osteogenicity: AnIn VitroMethod for Biomimetic Construct Assembly

Serino

D’Alessandro

Moscato

et al. 2013

Tissue Engineering Part C: Methods

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The traditional bone tissue-engineering approach exploits mesenchymal stem cells (MSCs) to be seeded once only on three-dimensional (3D) scaffolds, hence, differentiated for a certain period of time and resulting in a homogeneous osteoblast population at the endpoint. However, after achieving terminal osteodifferentiation, cell viability is usually markedly compromised. On the other hand, naturally occurring osteogenesis results from the coexistence of MSC progenies at distinct differentiative stages in the same microenvironment. This diversification also enables long-term viability of the mature tissue. We report an easy and tunable in vitro method to engineer simple osteogenic cell niches in a biomimetic fashion. The niches were grown via periodic reseeding of undifferentiated MSCs on MSC/scaffold constructs, the latter undergoing osteogenic commitment. Timefractioning of the seeded cell number during differentiation time of the constructs allowed graded osteogenic cell populations to be grown together on the same scaffolds (i.e., not only terminally differentiated osteoblasts). In such cell-dynamic systems, the overall differentiative stage of the constructs could also be tuned by varying the cell density seeded at each inoculation. In this way, we generated two different biomimetic niche models able to host good reservoirs of preosteoblasts and other osteoprogenitors after 21 culture days. At that time, the niche type resulting in 40.8% of immature osteogenic progenies and only 59.2% of mature osteoblasts showed a calcium content comparable to the constructs obtained with the traditional culture method (i.e., 100.03 -29.30 vs. 78.51 -28.50 pg/cell, respectively; p = not significant), the latter colonized only by fully differentiated osteoblasts showing exhausted viability. This assembly method for tissue-engineered constructs enabled a set of important parameters, such as viability, colonization, and osteogenic yield of the MSCs to be balanced on 3D scaffolds, thus achieving biomimetic in vitro models with graded osteogenicity, which are more complex and reliable than those currently used by tissue engineers.

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Section: Expansion and Culture Of Hmscsmentioning

confidence: 99%

Section: Osteogenic Differentiation Of the Niches: Qualitative And Sementioning

confidence: 99%

mentioning

confidence: 99%

mentioning

confidence: 99%

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Growing Bone Tissue-Engineered Niches with Graded Osteogenicity: AnIn VitroMethod for Biomimetic Construct Assembly

Serino

D’Alessandro

Moscato

et al. 2013

Tissue Engineering Part C: Methods

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“…At least five areas (including four corners and one center) were imaged and counted for each section. Cell viability was evaluated by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay as described previously [23] at 2, 7, and 14 days after seeding cells. The background absorption was subtracted after measuring the average OD of plain scaffolds without cells.…”

Section: Cell Survival and Viability Within Gs Scaffoldsmentioning

confidence: 99%

Graft of a Tissue-Engineered Neural Scaffold Serves as a Promising Strategy to Restore Myelination after Rat Spinal Cord Transection

Lai

Wang²,

Ling³

et al. 2014

Stem Cells and Development

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Remyelination remains a challenging issue in spinal cord injury (SCI). In the present study, we cocultured Schwann cells (SCs) and neural stem cells (NSCs) with overexpression of neurotrophin-3 (NT-3) and its high affinity receptor tyrosine kinase receptor type 3 (TrkC), respectively, in a gelatin sponge (GS) scaffold. This was aimed to generate a tissue-engineered neural scaffold and to investigate whether it could enhance myelination after a complete T10 spinal cord transection in adult rats. Indeed, many NT-3 overexpressing SCs (NT-3-SCs) in the GS scaffold assumed the formation of myelin. More strikingly, a higher incidence of NSCs overexpressing TrkC differentiating toward myelinating cells was induced by NT-3-SCs. By transmission electron microscopy, the myelin sheath showed distinct multilayered lamellae formed by the seeded cells. Eighth week after the scaffold was transplanted, some myelin basic protein (MBP)-positive processes were observed within the transplantation area. Remarkably, certain segments of myelin derived from NSC-derived myelinating cells and NT-3-SCs were found to ensheath axons. In conclusion, we show here that transplantation of the GS scaffold promotes exogenous NSC-derived myelinating cells and SCs to form myelins in the injury/transplantation area of spinal cord. These findings thus provide a neurohistological basis for the future application or transplantation using GS neural scaffold to repair SCI.

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A CD10‐OGP Membrane Peptolytic Signaling Axis in Fibroblasts Regulates Lipid Metabolism of Cancer Stem Cells via SCD1

et al. 2021

Advanced Science

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Carcinoma-associated fibroblasts (CAFs) consist of heterogeneous subpopulations that play a critical role in the dynamics of the tumor microenvironment. The extracellular signals of CAFs have been attributed to the extracellular matrix, cytokines, cell surface checkpoints, and exosomes. In the present study, it is demonstrated that the CD10 transmembrane hydrolase expressed on a subset of CAFs supports tumor stemness and induces chemoresistance. Mechanistically, CD10 degenerates an antitumoral peptide termed osteogenic growth peptide (OGP). OGP restrains the expression of rate-limiting desaturase SCD1 and inhibits lipid desaturation, which is required for cancer stem cells (CSCs). Targeting CD10 significantly improves the efficacy of chemotherapy in vivo. Clinically, CD10-OGP signals are associated with the response to neoadjuvant chemotherapy in patients with breast cancer. The collective data suggest that a nexus between the niche and lipid metabolism in CSCs is a promising therapeutic target for breast cancer.

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Gelatin/PLLA Sponge‐Like Scaffolds Allow Proliferation and Osteogenic Differentiation of Human Mesenchymal Stromal Cells

Cited by 25 publications

References 22 publications

Growing Bone Tissue-Engineered Niches with Graded Osteogenicity: AnIn VitroMethod for Biomimetic Construct Assembly

Growing Bone Tissue-Engineered Niches with Graded Osteogenicity: AnIn VitroMethod for Biomimetic Construct Assembly

Graft of a Tissue-Engineered Neural Scaffold Serves as a Promising Strategy to Restore Myelination after Rat Spinal Cord Transection

A CD10‐OGP Membrane Peptolytic Signaling Axis in Fibroblasts Regulates Lipid Metabolism of Cancer Stem Cells via SCD1

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