Assessment of in vitro aging of mesenchymal stem cell
,
Abstract
Mesenchyml stem cell (MSC) are receiving much attention in treatment of various diseases. The low frequency of MSCs in bone marrow (BM) necessitates their in vitro expansion prior to clinical use. We evaluated the effect of long term culture on the senescence of these cells. BM cells were taken from 11 transplant donors with mean age of 25 years. In different passages, MSC were examined for different aging indicators including: telomere length assay, differentiation ability, immunophenotyping of CD13, CD44 and CD34 antigens, determination of cumulative population dou¬blings (CPDs), and study of morphological characteristics of MSC cultures. The mean long term culture was 118 day and the mean passage number was 9. The average number of PD decreased from 7.7 to 1.2 in the 10th passage. The mean telomere length decreased from 9.19 Kbp to 8.7 kbp in the 9th passage. Differentiation potential dropped from the 6th passage on. The culture's morphological abnormalities were typical of the Hayflick model of cellular aging. We believe that MSC enter senescence almost undetectably from the moment of in vitro culturing. Si¬multaneously these cells are losing their stem cell characteristics. Therefore, it is much better to con¬sider them for cell and gene therapy early on.
Gerson SL. Mesenchymal stem cells: no longer second class marrow citizens. Nat Med. 1999 Mar;5(3):262-4
Anklesaria P, Kase K, Glowacki J. et al. Engraftment of a clonal bone marrow stromal cell line in vivo stimu- lates hematopoietic recovery from total body irradiation. Proc Natl Acad Sci U S A. 1987 Nov;84(21):7681-5
Koc ON, Gerson SL, Cooper BW, et al. Rapid hemato- poietic recovery after coinfusion of autologous-blood stem cells and culture-expanded marrow mesenchymal stem cells in advanced breast cancer patients receiving high-dose chemotherapy. J Clin Oncol. 2000 Jan;18(2):307-16.]
Devine SM, Hoffman R. Role of mesenchymal stem cells in hematopoietic stem cell transplantation. Curr Opin Hematol. 2000 Nov;7(6):358-63.
Bruder SP, Kurth AA, Shea M, Hayes WC, Jaiswal N, Kadiyala S. Bone regeneration by implantation of puri- fied, culture-expanded human mesenchymal stem cells. J Orthop Res. 1998 Mar;16(2):155-62.
Fukuda K, Sakamoto N, Narita T, et al. Application of efficient and specific gene transfer systems and organ culture techniques for the elucidation of mechanisms of epithelial-mesenchymal interaction in the developing gut. Dev Growth Differ. 2000 Jun;42(3):207-11.
Friedenstein AJ, Latzinik NW, Grosheva AG, Gor- skaya UF. Marrow microenvironment transfer by het- erotopic transplantation of freshly isolated and cultured cells in porous sponges. Exp Hematol. 1982 Feb;10(2):217-27.
Wexler SA, Donaldson C, Denning-Kendall P, Rice C, Bradley B, Hows JM. Adult bone marrow is a rich source of human mesenchymal 'stem' cells but umbilical cord and mobilized adult blood are not. Br J Haematol. 2003 Apr;121 (2):368-74.
Cheng FJ, Zou P, Zhong ZD, Guo R, Xiao J. [The growth characteristics of mesenchymal stem/progenitor cells in human umbilical cord blood] Zhongguo Shi Yan Xue Ye Xue Za Zhi. 2003 Dec;11(6):565-8.
Van Zglinicki T, SaretzkiG, Docke W, Lotze C. Mild hyperoxia shortens telomeres and inhibits prolifera- tion: a model for senescence? Exp.Cell Res. 1995;220:186.
Stenderup K, Justesen J, Clausen C, Kassem M. Ag- ing is associated with decreased maximal life span and accelerated senescence of bone marrow stromal cells. Bone. 2003 Dec;33(6):919-26.
Hayflick L. The limited in vitro life time of human diploid cell strain.Exp Cell Res. 1965 Mar;37:614-36.
Colter DC, Class R, DiGirolamo CM, Prockop DJ. Rapid expansion of recycling stem cells in cultures of plastic-adherent cells from human bone marrow. Proc Natl Acad Sci U S A. 2000 Mar 28;97(7):3213-8.
Smith JR, Lincoln DW 2nd. Aging of cells in cul- ture. Int Rev Cytol. 1984;89:151-77.
Allsopp RC, Harley CB. Evidence for a critical te- lomere length in senescent human fibroblasts. Exp Cell Res. 1995 Jul;219(1):130-6.
Kolquist KA, Ellisen LW, Counter CM, et al. Ex- pression of TERT in early premalignant lesions and a subset of cells in normal tissues. Nat Genet. 1998 Jun;19(2):182-6.
Baxter MA, Wynn RF, Jowitt SN, Wraith JE, Fair- bairn LJ, Bellantuono I. Study of telomere length reveals rapid aging of human marrow stromal cells following in vitro expansion. Stem Cells. 2004;22(5):675-82.
Parsch D, Fellenberg J, Brummendorf TH, Eschl- beck AM, Richter W. Telomere length and telomerase activity during expansion and differentiation of human mesenchymal stem cells and chondrocytes. J Mol Med. 2004 Jan;82(1):49-55. Epub 2003 Nov 28.
Pittenger MF, Mackay AM, Beck SC, et al. Multi- lineage potential of adult human mesenchymal stem cells. Science. 1999 Apr 2;284(5411):143-7.
Digirolamo CM, Stokes D, Colter D, Phinney DG, Class R, Prockop DJ. Propagation and senescence of hu- man marrow stromal cells in culture: a simple colony- forming assay identifies samples with the greatest poten- tial to propagate and differentiate. Br J Haematol. 1999 Nov;107(2):275-81.
Conget PA, Minguell JJ. Phenotypical and functional properties of human bone marrow mesenchymal progeni- tor cells. J Cell Physiol. 1999 Oct;181(1):67-73.
| Files | ||
| Issue | Vol 2, No 2 (2005) | |
| Section | Articles | |
| Keywords | ||
| #No Keywords# | ||
| Rights and permissions | |
|
This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License. |
