Abstract
Human adult stem cells (hASCs) offer a potentially renewable source of cell types that are easily isolated and rapidly expanded for use in regenerative medicine and cell therapies without the complicating ethical problems that are associated with embryonic stem cells. However, the eventual therapeutic use of hASCs requires that these cells and their derivatives maintain their genomic stability. There is currently a lack of systematic studies that are aimed at characterising aberrant chromosomal changes in cultured ASCs over time. However, the presence of mosaicism and accumulation of karyotypic abnormalities within cultured cell subpopulations have been reported. To investigate cytogenetic integrity of cultured human dental stem cell (hDSC) lines, we analysed four expanded hDSC cultures using classical G banding and fluorescent in situ hybridisation (FISH) with X chromosome specific probe. Our preliminary results revealed that about 70% of the cells exhibited karyotypic abnormalities including polyploidy, aneuploidy and ring chromosomes. The heterogeneous spectrum of abnormalities indicates a high frequency of chromosomal mutations that continuously arise upon extended culture. These findings emphasise the need for the careful analysis of the cytogenetic stability of cultured hDSCs before they can be used in clinical therapies.
Similar content being viewed by others
References
Andrews PW ((2004)) Author replay in comments. Nat Biotechnol 22:381–382
Benn PA (1977) Population kinetics of chromosomally abnormal human fibroblast subpopulations. Cytogenet Cell Genet 19:136–145
Bochkov NP, Voronina ES, Kosyakova NV, Liehr T, Rzhaninova AA, Katosova LD, Platonova VI, Gol’dshtein DV (2007) Chromosome variability of human multipotent mesenchymal stromal cells. Bull Exp Biol Med 143:122–126
Buzzard JJ, Gough NM, Crook JM, Colman A (2004) Karyotype of human ES cells during extended culture. Nat Biotechnol 22:381–382
Caisander G, Park H, Frej K, Lindqvist J, Bergh C, Lundin K, Hanson C (2006) Chromosomal integrity maintained in five human embryonic stem cell lines after prolonged in vitro culture. Chromosome Res 14:131–137
Carreira IM, Mascarenhas A, Matoso E, Couceiro AB, Ramos L, Dufke A, Mazauric M, Stressig R, Kosyakova N, Melo JB, Liehr T (2007) Three unusual but cytogenetically similar cases with up to five different cell lines involving structural and numerical abnormalities of chromosome 18. J Histochem Cytochem 55:1123–1128
Catalina P, Cobo F, Cortés JL, Nieto AI, Cabrera C, Montes R, Concha A, Menendez P (2007) Conventional and molecular cytogenetic diagnostic methods in stem cell research: a concise review. Cell Biol Int 31:861–869
Draper JS, Moore HD, Ruban LN, Gokhale PJ, Andrews PW (2004) Culture and characterization of human embryonic stem cells. Stem Cells Dev 13:325–336
Duailibi MT, Duailibi SE, Young CS, Bartlett JD, Vacanti JP, Yelick PC (2004) Bioengineered teeth from cultured rat tooth bud cells. J Dent Res 83:517
Duailibi MT, Duailibi SE, Duailibi Neto EF, Negreiros RM, Jorge WA, Ferreira LM, Vacanti JP, Yelick PC (2011) Tooth tissue engineering: optimal dental stem cell harvest based on tooth development. Artif Organs 35:129–135
Forsyth NR, Musio A, Vezzoni P, Simpson AH, Noble BS, McWhir J (2006) Physiologic oxygen enhances human embryonic stem cell clonal recovery and reduces chromosomal abnormalities. Cloning Stem Cells 8(1):16–23
Giraldo AM, Lynn JW, Godke RA, Bondioli KR (2006) Proliferative characteristics and chromosomal stability of bovine donor cells for nuclear transfer. Mol Reprod Dev 73:1230–1238
Hanson C, Caisander G (2005) Human embryonic stem cells and chromosome stability. APMIS 113:751–755
Hoffman LM, Carpenter MK (2005) Characterization and culture of human embryonic stem cells. Nat Biotechnol 23:699–708
Kaigler D, Mooney D (2001) Tissue engineering’s impact on dentistry. J Dent Educ 65:456–462
Ludwig TE, Bergendahl V, Levenstein ME, Yu J, Probasco MD, Thomson JA (2006) Feeder-independent culture of human embryonic stem cells. Nat Methods 3:637–646 (Erratum in: Nat Methods. 3(2006):867)
Maitra A, Arking DE, Shivapurkar N, Ikeda M, Stastny V, Kassauei K, Sui G, Cutler DJ, Liu Y, Brimble SN, Noaksson K, Hyllner J, Schulz TC, Zeng X, Freed WJ, Crook J, Abraham S, Colman A, Sartipy P, Matsui S, Carpenter M, Gazdar AF, Rao M, Chakravarti A (2005) Genomic alterations in cultured human embryonic stem cells. Nat Genet 37:1099–1103
Mastromonaco GF, Perrault SD, Betts DH, King WA (2006) Role of chromosome stability and telomere length in the production of viable cell lines for somatic cell nuclear transfer. BMC Dev Biol 6:41
Mikkola M, Olsson C, Palgi J, Ustinov J, Palomaki T, Horelli-Kuitunen N, Knuutila S, Lundin K, Otonkoski T, Tuuri T (2006) Distinct differentiation characteristics of individual human embryonic stem cell lines. BMC Dev Biol 6:40
Mitalipova MM, Rao RR, Hoyer DM, Johnson JA, Meisner LF, Jones KL, Dalton S, Stice SL (2005) Preserving the genetic integrity of human embryonic stem cells. Nat Biotechnol 23:19–20
Neff T, Beard BC, Kiem HP (2006) Survival of the fittest: in vivo selection and stem cell gene therapy. Blood 107:1751–1760
Rebuzzini P, Neri T, Mazzini G, Zuccotti M, Redi CA, Garagna S (2008) Karyotype analysis of the euploid cell population of a mouse embryonic stem cell line revealed a high incidence of chromosome abnormalities that varied during culture. Cytogenet Genome Res 121:18–24
Roschke AV, Stover K, Tonon G, Schäffer AA, Kirsch IR (2002) Stable karyotypes in epithelial cancer cell lines despite high rates of ongoing structural and numerical chromosomal instability. Neoplasia 4:19–31
Strelchenko N, Verlinsky O, Kukharenko V, Verlinsky Y (2004) Morula-derived human embryonic stem cells. Reprod Biomed Online 9:623–629
Suchánek J, Soukup T, Ivancaková R, Karbanová J, Hubková V, Pytlík R, Kucerová L (2007) Human dental pulp stem cells–isolation and long term cultivation. Acta Medica (Hradec Kralove) 50:195–201
Vorsanova SG, Yurov YB, Iororv IY (2010) Human interphase chromosomes: a review of available molecular cytogenetic technologies. Mol Cytogenet 11(3):1
Wang Y, Huso DL, Harrington J, Kellner J, Jeong DK, Turney J, McNiece IK (2005) Outgrowth of a transformed cell population derived from normal human BM mesenchymal stem cell culture. Cytotherapy 7:509–519
Young CS, Terada S, Vacanti JP, Honda M, Bartlett JD, Yelick PC (2002) Tissue engineering of complex tooth structures on biodegradable polymer scaffolds. J Dent Res 81:695–700
Young CS, Abukawa H, Asrican R, Ravens M, Troulis MJ, Kaban LB, Vacanti JP, Yelick PC (2005) Tissue-engineered hybrid tooth and bone. Tissue Eng 11:1599–1610
Zhang ZX, Guan LX, Zhang K, Wang S, Cao PC, Wang YH, Wang Z, Dai LJ (2007) Cytogenetic analysis of human bone marrow-derived mesenchymal stem cells passaged in vitro. Cell Biol Int 31:645–648
Acknowledgments
We wish to thank the INCT- Biofabrication Institute, CNPq 573661/2008-1, FAPESP 08/57860-3, and the Rede Biofab, Ibero-American Network of Biofabrication—BIOFAB-CYTED (208RT0340). This work was supported by FAPESP grants 07- 58856-7, 07-51227-4, 07-59488-1, and NIH/NIDCR R01DE016132 (PCY), and R03TW007665 (PCY, MTD, SED) awards. We are also grateful to Dr. Waldyr Antonio Jorge, Ph.D., Oral Maxillofacial Surgery Course of Dental School, FFO–FOUSP Professor of University of São Paulo for contribution of the biological material access.
Author information
Authors and Affiliations
Corresponding author
Additional information
Monica Talarico Duailibi, Leslie Domenici Kulikowski and Silvio Eduardo Duailibi contributed equally to this manuscript.
Rights and permissions
About this article
Cite this article
Duailibi, M.T., Kulikowski, L.D., Duailibi, S.E. et al. Cytogenetic instability of dental pulp stem cell lines. J Mol Hist 43, 89–94 (2012). https://doi.org/10.1007/s10735-011-9373-z
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10735-011-9373-z