A multiplex PCR technique to characterize human bone marrow derived mesenchymal stem cells

Rallapalli, Suneel; Bishi, Dillip Kumar; Verma, Rama Shanker; Cherian, Kotturathu Mammen; Guhathakurta, Soma

doi:10.1007/s10529-009-0106-2

Cited by 13 publications

(8 citation statements)

References 24 publications

(23 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…This suggests that the expression pes se of typical CD markers used for MSC characterization is not conclusive for defining “true” stemness and should be used in tandem with other assays. This has also been highlighted in the case of adult BM-MSCs [46]. For ADSC the main phenotypic changes related to increase in size, partial change in morphology, and a decline in clonogenicity.…”

Section: Discussionmentioning

confidence: 83%

Comparative Evaluation of Human Mesenchymal Stem Cells of Fetal (Wharton's Jelly) and Adult (Adipose Tissue) Origin during ProlongedIn VitroExpansion: Considerations for Cytotherapy

Christodoulou

Kolisis

Papaevangeliou³

et al. 2013

Stem Cells International

100

View full text Add to dashboard Cite

Mesenchymal stem cells (MSCs) are somatic cells with a dual capacity for self-renewal and differentiation, and diverse therapeutic applicability, both experimentally and in the clinic. These cells can be isolated from various human tissues that may differ anatomically or developmentally with relative ease. Heterogeneity due to biological origin or in vitro manipulation is, nevertheless, considerable and may equate to differences in qualitative and quantitative characteristics which can prove crucial for successful therapeutic use. With this in mind, in the present study we have evaluated the proliferation kinetics and phenotypic characteristics of MSCs derived from two abundant sources, that is, fetal umbilical cord matrix (Wharton's jelly) and adult adipose tissue (termed WJSC and ADSC, resp.) during prolonged in vitro expansion, a process necessary for obtaining cell numbers sufficient for clinical application. Our results show that WJSC are derived with relatively high efficiency and bear a substantially increased proliferation capacity whilst largely sustaining the expression of typical immunophenotypic markers, whereas ADSC exhibit a reduced proliferation potential showing typical signs of senescence at an early stage. By combining kinetic with phenotypic data we identify culture thresholds up to which both cell types maintain their stem properties, and we discuss the practical implications of their differences.

show abstract

Section: Discussionmentioning

confidence: 83%

Comparative Evaluation of Human Mesenchymal Stem Cells of Fetal (Wharton's Jelly) and Adult (Adipose Tissue) Origin during ProlongedIn VitroExpansion: Considerations for Cytotherapy

Christodoulou

Kolisis

Papaevangeliou³

et al. 2013

Stem Cells International

100

View full text Add to dashboard Cite

show abstract

“…Moreover, most of the cell surface markers utilized to sort subpopulations of human MSC by flow cytometry have not been validated in sheep [21]. Gene expression-based technologies may be useful for the identification of possible molecules described as MSC markers [31,32]. In our study, RT-qPCR was performed to quantify the mRNA expression levels of six cell surface antigens considered as either positive ( CD29 , CD73 , CD90 and CD105 ) or negative ( CD34 and CD45 ) MSC markers in humans.…”

Section: Discussionmentioning

confidence: 99%

Isolation and characterization of ovine mesenchymal stem cells derived from peripheral blood

et al. 2012

View full text Add to dashboard Cite

BackgroundMesenchymal stem cells (MSCs) are multipotent stem cells with capacity to differentiate into several mesenchymal lineages. This quality makes MSCs good candidates for use in cell therapy. MSCs can be isolated from a variety of tissues including bone marrow and adipose tissue, which are the most common sources of these cells. However, MSCs can also be isolated from peripheral blood. Sheep has been proposed as an ideal model for biomedical studies including those of orthopaedics and transmissible spongiform encephalopathies (TSEs). The aim of this work was to advance these studies by investigating the possibility of MSC isolation from ovine peripheral blood (oPB-MSCs) and by subsequently characterizing there in vitro properties.ResultsPlastic-adherent fibroblast-like cells were obtained from the mononuclear fraction of blood samples. These cells were analysed for their proliferative and differentiation potential into adipocytes, osteoblasts and chondrocytes, as well as for the gene expression of cell surface markers. The isolated cells expressed transcripts for markers CD29, CD73 and CD90, but failed to express the haematopoietic marker CD45 and expressed only low levels of CD105. The expression of CD34 was variable. The differentiation potential of this cell population was evaluated using specific differentiation media. Although the ability of the cultures derived from different animals to differentiate into adipocytes, osteoblasts and chondrocytes was heterogeneous, we confirmed this feature using specific staining and analysing the gene expression of differentiation markers. Finally, we tested the ability of oPB-MSCs to transdifferentiate into neuronal-like cells. Morphological changes were observed after 24-hour culture in neurogenic media, and the transcript levels of the neurogenic markers increased during the prolonged induction period. Moreover, oPB-MSCs expressed the cellular prion protein gene (PRNP), which was up-regulated during neurogenesis.ConclusionsThis study describes for the first time the isolation and characterization of oPB-MSCs. Albeit some variability was observed between animals, these cells retained their capacity to differentiate into mesenchymal lineages and to transdifferentiate into neuron-like cells in vitro. Therefore, oPB-MSCs could serve as a valuable tool for biomedical research in fields including orthopaedics or prion diseases.

show abstract

“…Whether the decrease in surface expression of CD73 in TDSCs (CI) was associated with reduced migration ability of these cells needs further research. The reduced mRNA expression of CD73 was also reported to positively correlate with the reduced proliferation and differentiation efficiency of human MSCs during passaging in vitro [30]. Hence, the reduction in the surface expression of CD73 might also be explained by the committed differentiation of TDSCs (CI) to the chondrogenic and osteogenic lineages.…”

mentioning

confidence: 98%

Altered Fate of Tendon-Derived Stem Cells Isolated from a Failed Tendon-Healing Animal Model of Tendinopathy

Rui

Lui

Wong

et al. 2013

Stem Cells and Development

View full text Add to dashboard Cite

We hypothesized that altered fate of tendon-derived stem cells (TDSCs) might contribute to chondro-ossification and failed healing in the collagenase-induced (CI) tendon injury model. This study aimed to compare the yield, proliferative capacity, immunophenotypes, senescence, and differentiation potential of TDSCs isolated from healthy (HT) and CI tendons. TDSCs were isolated from CI and healthy Sprague-Dawley rat patellar tendons. The yield, proliferative capacity, immunophenotypes, and senescence of TDSCs (CI) and TDSCs (HT) were compared by colony-forming unit assay, BrdU assay, flow cytometry, and b-galactosidase activity assay, respectively. Their osteogenic and chondrogenic differentiation potentials and mRNA expression of tendon-related markers were compared using standard assays. More TDSCs, which showed a lower proliferative potential and a higher cellular senescence were present in the CI patellar tendons compared to HT tendons. There was a higher alkaline phosphatase activity and mineralization in TDSCs (CI) in both basal and osteogenic media. More chondrocyte-like cells and higher proteoglycan deposition, Sox9 and collagen type II expression were observed in TDSCs (CI) pellets upon chondrogenic induction. There was a higher protein expression of Sox9, but a lower mRNA expression of Col1a1, Scx, and Tnmd in TDSCs (CI) in a basal medium. In conclusion, TDSCs (CI) showed altered fate, a higher cellular senescence, but a lower proliferative capacity compared to TDSCs (HT), which might contribute to pathological chondro-ossification and failed tendon healing in this animal model.

show abstract

A multiplex PCR technique to characterize human bone marrow derived mesenchymal stem cells

Cited by 13 publications

References 24 publications

Comparative Evaluation of Human Mesenchymal Stem Cells of Fetal (Wharton's Jelly) and Adult (Adipose Tissue) Origin during ProlongedIn VitroExpansion: Considerations for Cytotherapy

Comparative Evaluation of Human Mesenchymal Stem Cells of Fetal (Wharton's Jelly) and Adult (Adipose Tissue) Origin during ProlongedIn VitroExpansion: Considerations for Cytotherapy

Isolation and characterization of ovine mesenchymal stem cells derived from peripheral blood

Altered Fate of Tendon-Derived Stem Cells Isolated from a Failed Tendon-Healing Animal Model of Tendinopathy

Contact Info

Product

Resources

About