Isolation of umbilical cord mesenchymal stem cells using human blood derivatives accompanied with explant method

Hassan, Ghmkin; Kasem, Issam; Antaki, Reham; Mohammad, Mohammad Bahjat; AlKadry, Ranad; Aljamali, Majd

doi:10.21037/sci.2019.08.06

Cited by 15 publications

(9 citation statements)

References 23 publications

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“…Mesenchymal Stromal Cells (MSCs) are non-hematopoietic multipotent cells which can be isolated from different tissues from adult, perinatal and fetal samples [1,2]. Some sources are adipose tissue [3], bone marrow [4], umbilical cord Wharton's jelly and blood [5,6], periosteum [7], skin [8], amniotic fluid [9] and the placenta [10]. These cells have the capability to differentiate into a variety of different mesenchymal lineage cells such as osteoblasts, chondrocytes, adipocytes, fibroblasts and myoblasts [2].…”

Section: Introductionmentioning

confidence: 99%

Biodistribution of Mesenchymal Stromal Cells after Administration in Animal Models and Humans: A Systematic Review

Sánchez‐Díaz

Quiñones-Vico

Torre

et al. 2021

JCM

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Mesenchymal Stromal Cells (MSCs) are of great interest in cellular therapy. Different routes of administration of MSCs have been described both in pre-clinical and clinical reports. Knowledge about the fate of the administered cells is critical for developing MSC-based therapies. The aim of this review is to describe how MSCs are distributed after injection, using different administration routes in animal models and humans. A literature search was performed in order to consider how MSCs distribute after intravenous, intraarterial, intramuscular, intraarticular and intralesional injection into both animal models and humans. Studies addressing the biodistribution of MSCs in “in vivo” animal models and humans were included. After the search, 109 articles were included in the review. Intravenous administration of MSCs is widely used; it leads to an initial accumulation of cells in the lungs with later redistribution to the liver, spleen and kidneys. Intraarterial infusion bypasses the lungs, so MSCs distribute widely throughout the rest of the body. Intramuscular, intraarticular and intradermal administration lack systemic biodistribution. Injection into various specific organs is also described. Biodistribution of MSCs in animal models and humans appears to be similar and depends on the route of administration. More studies with standardized protocols of MSC administration could be useful in order to make results homogeneous and more comparable.

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Section: Introductionmentioning

confidence: 99%

Biodistribution of Mesenchymal Stromal Cells after Administration in Animal Models and Humans: A Systematic Review

Sánchez‐Díaz

Quiñones-Vico

Torre

et al. 2021

JCM

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“…In contrast, the DT for cells grown in XM remained steady and slowly increased from 15 h at P3 to 27 h at P40. Several published studies showed a signi cant increase in DT of adult and umbilical cord-derived MSCs upon passaging in FM compared to XM (20)(21)(22). In addition, the CFE of pMSCs was signi cantly lower in FM compared to XM.…”

Section: Discussionmentioning

confidence: 96%

“…Several reports describe xeno-free medium (XM) used to culture MSCs (15)(16)(17)(18)(19). The XM shows more promise in maintaining the selfrenewability of MSCs than FM (20)(21)(22). Despite the problems with maintaining MSCs during propagation, molecular changes limiting the self-renewal and differentiation potential upon passaging or culturing are poorly understood.…”

Section: Introductionmentioning

confidence: 99%

WNT and VEGF/PDGF signaling regulate self-renewal in primitive mesenchymal stem cells

Mazzella

Walker

Cormier

et al. 2023

Preprint

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Background Therapeutic use of multipotent mesenchymal stem cells (MSCs) is hampered due to poor growth and limited self-renewal potential. The self-renewal potential of MSCs is also affected during propagation and changes are poorly understood. This study investigated the molecular mechanism involved in the self-renewal of primitive (p) MSCs. Methods pMSCs were cultured to low passage (LP), P3, and high passage (HP), P20, in fetal bovine serum medium (FM) and xeno-free medium (XM). The characteristics of LP and HP pMSCs were evaluated for morphology, expression of cell surface markers, doubling time (DT), colony forming efficiency (CFE), proliferation by BrdU assay, telomerase activity and trilineage differentiation. We then examined transcriptome and nucleosome occupancies using RNA-seq and MNase-seq, respectively analyses. Results pMSCs grown in FM gradually changed morphology to large elongated cells and showed a significant reduction in the expression of CD90 and CD49f, CFE, proliferation, and telomerase activity. In addition, cells had a greater propensity to differentiate into the adipogenic lineage. In contrast, pMSCs grown in XM maintained small fibroblastoid morphology, self-renewal, and differentiation potential. Transcriptomic analysis showed upregulation of genes involved in self-renewal, cell cycle, and DNA replication in XM-grown pMSCs. Whereas senescence genes were upregulated in cells in FM. MNase-seq analysis revealed less nucleosomal occupancies in self-renewal genes and senescence genes in pMSCs grown in XM and FM, respectively. The expression of selected genes associated with self-renewal, cell cycle, DNA replication, differentiation, and senescence was confirmed by qRT-PCR. These results led us to propose signaling pathways involved in the self-renewal and senescence of pMSCs. Conclusion We conclude that the self-renewal potential of pMSCs is controlled by WNT and VEGF/PDGF, but TGFβ and PI3K signaling induce senescence.

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“…In response to these challenges, xeno-free medium (XM) has emerged as a promising alternative for MSC culture [ 17 – 21 ]. XM comprises human serum and proteins, offering potential advantages in maintaining MSC self-renewal [ 22 – 24 ].…”

Section: Introductionmentioning

confidence: 99%

Regulation of self-renewal and senescence in primitive mesenchymal stem cells by Wnt and TGFβ signaling

Mazzella,

Walker,

Cormier

et al. 2023

Stem Cell Res Ther

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Background The therapeutic application of multipotent mesenchymal stem cells (MSCs) encounters significant challenges, primarily stemming from their inadequate growth and limited self-renewal capabilities. Additionally, as MSCs are propagated, their ability to self-renew declines, and the exact cellular and molecular changes responsible for this are poorly understood. This study aims to uncover the complex molecular mechanisms that govern the self-renewal of primitive (p) MSCs. Methods We grew pMSCs using two types of medium, fetal bovine serum (FM) and xeno-free (XM), at both low passage (LP, P3) and high passage (HP, P20). To evaluate LP and HP pMSCs, we examined their physical characteristics, cell surface markers, growth rate, colony-forming ability, BrdU assays for proliferation, telomerase activity, and potential to differentiate into three lineages. Moreover, we conducted RNA-seq to analyze their transcriptome and MNase-seq analysis to investigate nucleosome occupancies. Results When grown in FM, pMSCs underwent changes in their cellular morphology, becoming larger and elongated. This was accompanied by a decrease in the expression of CD90 and CD49f, as well as a reduction in CFE, proliferation rate, and telomerase activity. In addition, these cells showed an increased tendency to differentiate into the adipogenic lineage. However, when grown in XM, pMSCs maintained their self-renewal capacity and ability to differentiate into multiple lineages while preserving their fibroblastoid morphology. Transcriptomic analysis showed an upregulation of genes associated with self-renewal, cell cycle regulation, and DNA replication in XM-cultured pMSCs, while senescence-related genes were upregulated in FM-cultured cells. Further analysis demonstrated differential nucleosomal occupancies in self-renewal and senescence-related genes for pMSCs grown in XM and FM, respectively. These findings were confirmed by qRT-PCR analysis, which revealed alterations in the expression of genes related to self-renewal, cell cycle regulation, DNA replication, differentiation, and senescence. To understand the underlying mechanisms, we investigated the involvement of Wnt and TGFβ signaling pathways by modulating them with agonists and antagonists. This experimental manipulation led to the upregulation and downregulation of self-renewal genes in pMSCs, providing further insights into the signaling pathways governing the self-renewal and senescence of pMSCs. Conclusion Our study shows that the self-renewal potential of pMSCs is associated with the Wnt pathway, while senescence is linked to TGFβ.

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Isolation of umbilical cord mesenchymal stem cells using human blood derivatives accompanied with explant method

Cited by 15 publications

References 23 publications

Biodistribution of Mesenchymal Stromal Cells after Administration in Animal Models and Humans: A Systematic Review

Biodistribution of Mesenchymal Stromal Cells after Administration in Animal Models and Humans: A Systematic Review

WNT and VEGF/PDGF signaling regulate self-renewal in primitive mesenchymal stem cells

Regulation of self-renewal and senescence in primitive mesenchymal stem cells by Wnt and TGFβ signaling

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