Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2008 Dec;41(6):909-922.
doi: 10.1111/j.1365-2184.2008.00559.x.

Study of oncogenic transformation in ex vivo expanded mesenchymal cells, from paediatric bone marrow

D M Choumerianou  1 , H Dimitriou  1 , C Perdikogianni  1 , G Martimianaki  1 , M Riminucci  1 , M Kalmanti  1
Affiliations

Study of oncogenic transformation in ex vivo expanded mesenchymal cells, from paediatric bone marrow

D M Choumerianou et al. Cell Prolif. 2008 Dec.

Abstract

Objectives: Mesenchymal stromal cells (MSCs) have attracted considerable interest in both the scientific and clinical fields. In order to obtain a sufficient cell number for application, their in vitro expansion is necessary, but during this process their characteristics may be altered and cells may acquire oncogenic properties. We have investigated properties of MSC that may be related to oncogenesis, a critical parameter that has to be evaluated prior to MSC clinical use.

Materials and methods: We studied the expression of p53, p16, RB, H-RAS and human telomerase reverse transcriptase (hTERT) in MSCs from bone marrow of children diagnosed with idiopathic thrombocytopenic purpura (ITP) and autoimmune neutropenia. The same cells were seeded in soft agar to confirm their anchorage dependence and were karyotypically analysed. Finally, MSCs were subcutaneously transplanted into SCID mice and their ectopic osteogenic as well as tumorigenic potential was evaluated.

Results: We have shown that MSCs derived from bone marrow of children with ITP and autoimmune neutropenia do not undergo transformation, the cells expressed normal levels of p53, p16, RB and H-RAS. Expression of hTERT was undetectable, chromosome content remained stable, and their anchorage dependence was confirmed. In an in vivo model, when MSCs were subcutaneously transplanted into SCID mice, no tumorigenesis was observed.

Conclusions: These findings suggest that MSCs from bone marrow of children do not have oncogenic properties and, therefore, represent validate candidates for applications in regenerative medicine.

PubMed Disclaimer

Figures

Figure 1
Figure 1
Representative electrophoresis results of RT‐PCR products for the genes p53, p16, RB and H‐RAS. Analysis was performed at different mesenchymal stromal cell (MSC) passages of ITP and AIN samples. On the first row, the bands correspond to the gene of interest and on the second row to GAPDH.
Figure 2
Figure 2
mRNA expression of p53, p16, RB, H‐RAS relative to GAPDH in mesenchymal stromal cells (MSC) according to disease group. Bar graphs represent mean expression levels of three independent experiments with SD (error bars). The expression levels of the same genes in HUVEC cells are denoted with a dot.
Figure 3
Figure 3
Culture of mesenchymal stromal cells (MSCs) on soft agar for the assay of anchorage dependence. Pictures were taken on day 7 (a, b, c) and 14 (d, e, f) and correspond to ΗL‐60 (c, f), P2 MSCs (a, d) and P6 MSCs (b, e). No colonies were observed in MSC samples in contrast with HL‐60 cells where foci were visible from day 7.
Figure 4
Figure 4
Electrophoresis of human telomerase reverse transcriptase (hTERT) real‐time PCR products. Four mesenchymal stromal cell (MSC) samples are shown at P2 and P6. hTERT is expressed in the cell line M5 but not in the MSC samples.
Figure 5
Figure 5
Representative G‐banding karyotype results. P2 MSCs (a) and P6 MSCs (b) from two different male patients. All karyotypes were normal. MSC, mesenchymal stromal cell.
Figure 6
Figure 6
Histochemically stained differentiated mesenchymal stromal cells. (a) adipocytes stained with oil red O, (b) osteocytes stained with von Kossa and (c) chondrocytes stained with Alcian Blue.
Figure 7
Figure 7
Mesenchymal stromal cell cultured with (a) and without fibroblast growth factor‐2 (b). Immunophenotype analysis results before subcutaneous transplantation into SCID mice are presented.
Figure 8
Figure 8
Histological analysis. Deposition of bone matrix onto the carrier in transplants of mesenchymal stromal cell (MSC) grown with fibroblast growth factor (FGF)‐2 (b, c) is shown. Only fibrous tissue and hydroxyapatite/tricalcium phosphate (HA/TCP) particles were detected in transplants of untreated cells (a). None of the samples showed evidence of tumour growth.
See this image and copyright information in PMC

Similar articles

See all similar articles

Cited by

See all "Cited by" articles

References

    1. Bang OY, Lee JS, Lee PH, Lee G (2005) Autologous mesenchymal stem cell transplantation in stroke patients. Ann. Neurol. 57, 874–882. - PubMed
    1. Barry FP (2003) Mesenchymal stem cell therapy in joint disease. Novartis Found. Symp. 249, 86–96. - PubMed
    1. Bernardo ME, Zaffaroni N, Novara F, Cometa AM, Avanzini MA, Moretta A, Montagna D, Maccario R, Villa R, Daidone MG, Zuffardi O, Locatelli F (2007) Human bone marrow derived mesenchymal stem cells do not undergo transformation after long‐term in vitro culture and do not exhibit telomere maintenance mechanisms. Cancer Res. 67, 9142–9149. - PubMed
    1. Bianco P, Kuznetsov SA, Riminucci M, Fisher LW, Spiegel AM, Robey PG (1998) Reproduction of human fibrous dysplasia of bone in immunocompromised mice by transplanted mosaics of normal and Gsalpha‐mutated skeletal progenitor cells. J. Clin. Invest. 101, 1737–1744. - PMC - PubMed
    1. Campagnolli C, Bellantuonno I, Kumar S, Fairnairn LJ, Roberts I, Fisk NM (2002) High transduction efficiency of circulating first trimester fetal mesenchymal cells: potential targets for in utero ex vivo gene therapy. BJOG 109, 952–954. - PubMed

Publication types

MeSH terms