Diet composition transiently modulates proliferative and potency features of human cord blood-derived mesenchymal stem cells

Ragni, Enrico; Parazzi, Valentina; Crosti, Mariacristina; Moro, Monica; Giordano, Rosaria; Lazzari, Lorenza

doi:10.1016/j.biocel.2014.09.017

Cited by 6 publications

(6 citation statements)

References 60 publications

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“…Although no statistical significance was observed, the ratio seemed to be lower for CB-MSC 1, as shown by both the mean and median, but concomitantly also presented the greatest degree of variation, as shown by SD and minimum and maximum values. This was consistent with previously reported data 12 , comparing CB-MSC morphology of the same population in two different media. No significant differences in cell morphology were observed among CB-MSC 1, 2, and 3 or between LL-CB-MSCs and CB-MSC 2 or 3 (data not shown).…”

Section: Resultssupporting

confidence: 94%

“…The idea was to offer the best environment to facilitate CB mesenchymal progenitor attachment to a plastic surface, which is considered essential for their survival 11 , and also to better support the generation and growth of stromal colonies. Second, considering the great influence of the medium composition on MSC stemness and other biological features 12 , we sought to further evaluate the peculiar characteristics of the CB-MSCs cultured under these new conditions to provide more complete information about their effect and on their properties. Finally, we wanted to investigate if such an optimal culture environment could be applied to rescue the SL-CB-MSC phenotype and induce them to adopt the same clinically relevant proliferative properties demonstrated by LL-CB-MSCs in standard culture conditions.…”

Section: Introductionmentioning

confidence: 99%

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A Chemically Defined Medium-Based Strategy to Efficiently Generate Clinically Relevant Cord Blood Mesenchymal Stromal Colonies

et al. 2016

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During the last decade it has been demonstrated that mesenchymal progenitors are present and can be isolated also from cord blood (CB). Recently, we managed to set up a standard protocol allowing the isolation of mesenchymal stromal cells (MSCs) with high proliferative potential and multiple differentiation capabilities, whereas the generation rate of MSC-initiating colonies could still be further improved. Herein, we strikingly succeeded in defining some simple and basic culture conditions based on the use of a chemically defined medium that increased the colony isolation efficiency up to almost 80% of processed CB units. Importantly, this result was achieved irrespective of CB unit white blood cell content and time elapsed from delivery, two limiting parameters involved with processing CB units. Thus, this high efficiency is guaranteed without strict selection of the starting material. In addition, since we are profoundly concerned about how different culture conditions can influence cell behavior, we devoted part of this study to in-depth characterization of the established CB-MSC populations to confirm their stemness features in this novel isolation and culture system. Therefore, an extended study of their immunophenotype, including classical pericytic markers, and a detailed molecular analysis addressing telomere length and also stemness-related microRNA contribution were performed. In summary, we propose a straightforward, extremely efficient, and reliable approach to isolate and expand thoroughly characterized CB-MSCs, even when poor-quality CB units are the only available source, or there is no space for an isolation to fail.

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Section: Resultssupporting

confidence: 94%

Section: Introductionmentioning

confidence: 99%

A Chemically Defined Medium-Based Strategy to Efficiently Generate Clinically Relevant Cord Blood Mesenchymal Stromal Colonies

et al. 2016

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“…Antiproliferative effects have also been described for miR-152 [68], miR-125b [69], miR-26a [70], miR-27b [71], and miR-143 and miR-145 [72]. Notably, many of the MSC miRNAs had already been identified to be MSC markers, confirming the robustness of the profiling analysis and of these miRNAs in defining MSC properties [73].…”

Section: Discussionmentioning

confidence: 66%

Using miRNA-mRNA Interaction Analysis to Link Biologically Relevant miRNAs to Stem Cell Identity Testing for Next-Generation Culturing Development

Crabbé

Gijbels²,

Visser³

et al. 2016

Stem Cells Translational Medicine

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Therapeutic benefit of stem cells has been demonstrated in multiple disease models and clinical trials. Robust quality assurance is imperative to make advancements in culturing procedures to enable large-scale cell manufacturing without hampering therapeutic potency. MicroRNAs (miRNAs or miRs) are shown to be master regulators of biological processes and are potentially ideal quality markers. We determined miRNA markers differentially expressed under nonclinical multipotent adult progenitor cell (MAPC) and mesenchymal stem cell (MSC) culturing conditions that regulate important stem cell features, such as proliferation and differentiation. These bone marrow-derived stem cell types were selected because they both exert therapeutic functions, but have different proliferative and regenerative capacities. To determine cell-specific marker miRNAs and assess their effects on stem cell qualities, a miRNA and mRNA profiling was performed on MAPCs and MSCs isolated from three shared donors. We applied an Ingenuity Pathway Analysis-based strategy that combined an integrated RNA profile analysis and a biological function analysis to determine the effects of miRNA-mRNA interactions on phenotype. This resulted in the identification of important miRNA markers linked to cell-cycle regulation and development, the most distinctive being MAPC marker miR-204-5p and MSC marker miR-335-5p, for which we provide in vitro validation of its function in differentiation and cell cycle regulation, respectively. Importantly, marker expression is maintained under xeno-free conditions and during bioreactor isolation and expansion of MAPC cultures. In conclusion, the identified biologically relevant miRNA markers can be used to monitor stem cell stability when implementing variations in culturing procedures. STEM CELLS TRANSLATIONAL MEDICINE 2016;5:709-722 SIGNIFICANCEHuman adult marrow stromal stem cells have shown great potential in addressing unmet health care needs. Quality assurance is imperative to make advancements in large-scale manufacturing procedures. MicroRNAs are master regulators of biological processes and potentially ideal quality markers. MicroRNA and mRNA profiling data of two human adult stem cell types were correlated to biological functions in silico. Doing this provided evidence that differentially expressed microRNAs are involved in regulating specific stem cell features. Furthermore, expression of a selected microRNA panel was maintained in next-generation culturing platforms, demonstrating the robustness of microRNA profiling in stem cell comparability testing.

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“…Specific primers overlapping two exons in order to avoid amplification due to genomic DNA contamination were designed based on GenBank (Table ) for NPT2A, NPT2C, PIT1, PIT2, BGLAP, and NHERF1. Primers for GAPDH, COL1A1, ATF4, and RUNX2 were previously described . All reactions were performed in triplicate and media of C T values was calculated.…”

Section: Methodsmentioning

confidence: 99%

“…Primers for GAPDH, COL1A1, ATF4, and RUNX2 were previously described. 27,28 All reactions were performed in triplicate and media of C T values was calculated. Target gene expression was normalized to the expression of the housekeeping gene glyceraldehyde-3-phosphate dehydrogenase (GAPDH) using the Bio-Rad CFX Manager Software.…”

Section: Quantitative Real-time Rt-pcrmentioning

confidence: 99%

Suppressive effects of tenofovir disoproxil fumarate, an antiretroviral prodrug, on mineralization and type II and type III sodium‐dependent phosphate transporters expression in primary human osteoblasts

Barbieri

Chiodini

Ragni

et al. 2018

J of Cellular Biochemistry

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Tenofovir disoproxil fumarate (TDF) is an antiretroviral drug commonly used for the management of Human Immunodeficiency Virus (HIV) in highly active antiretroviral therapy (HAART) and of chronic Hepatitis B Virus (HBV) infections. Long-term TDF-treated subjects present decrease of bone mineral density and rarely severe osteomalacia. Although these adverse effects have been attributed to the impaired proximal tubule function, a possible direct involvement of TDF on osteoblasts should be taken into account. The aim of this study was to evaluate whether sodium phosphate transporters NPT2A (sodium-dependent phosphate transport protein 2A), NPT2C (sodium-dependent phosphate transport protein 2C), PIT1 (sodium-dependent phosphate transporter 1), and PIT2 (sodium-dependent phosphate transporter 2) were expressed in primary human osteoblasts (HOBs), whether their expression was related to HOBs differentiation and whether TDF could affect mineralization and gene expression. PIT1 and PIT2 were expressed under proliferating conditions and increased after induction of mineralization, while NPT2A and NPT2C were almost undetectable. In HOBs TDF exposure induced a significant dose-dependent decrease in mineralization. Moreover, TDF caused a reduction of COL1A1 and of ATF4 expression in differentiated HOBs. In summary, HOBs do not express NPT2A and NPT2C and do express PIT1 and PIT2, suggesting a role of these two latter in human osteoblast mineralization. TDF impairs osteoblast mineralization, confirming a direct negative effect on bone. Therefore, in clinical practice, bone damage must be suspected and evaluated also in patients receiving TDF without kidney function alterations.

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Diet composition transiently modulates proliferative and potency features of human cord blood-derived mesenchymal stem cells

Cited by 6 publications

References 60 publications

A Chemically Defined Medium-Based Strategy to Efficiently Generate Clinically Relevant Cord Blood Mesenchymal Stromal Colonies

A Chemically Defined Medium-Based Strategy to Efficiently Generate Clinically Relevant Cord Blood Mesenchymal Stromal Colonies

Using miRNA-mRNA Interaction Analysis to Link Biologically Relevant miRNAs to Stem Cell Identity Testing for Next-Generation Culturing Development

Suppressive effects of tenofovir disoproxil fumarate, an antiretroviral prodrug, on mineralization and type II and type III sodium‐dependent phosphate transporters expression in primary human osteoblasts

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