Büyüme faktörlerinin kordon kanı kök hücre içeriği üzerine etkisinin hücrelerde Rodamin123 birikimi ile belirlenmesi

Abstract

Amaç: Mitokondri zarında toplanan rodamin123 Rd123 , hücrede mitokondri sayısı veya aktivitesi hakkında bilgi vermekte ve kök hücreyi fonksiyonel açıdan tanımlamada bir belirteç olarak kullanılmaktadır. Kök hücreler metabolik olarak inaktif veya uyku fazında olduklarından, bu hücrelerde Rd123 birikimi oldukça düşük düzeydedir. Çalışmamızda, büyüme faktörlerinin kordon kanı KK kök hücre içeriği üzerine etkisini araştırmak için, in vitro kültür periyodunda hücrelerin Rd123 alımını belirlemeyi amaçladık. Gereç Ve Yöntem: Sağlıklı ve zamanında doğum yapan 15 gönüllüden KK örnekleri alındı. Mononükleer hücreler ayrıldıktan sonra interlökin-3 IL-3 , IL-6, IL-11, stem cell faktör SCF , flt3/flk2 ligand FL ve trombopoetin TPO ile 14 gün inkübe edildi. Kültür öncesi, kültürün 7. ve 14. günlerinde hücreler alınarak Rd123 alım düzeyleri flow sitometride analiz edildi. Sonuçlar: Kök hücre üzerine etkili büyüme faktörleri ile yapılan kısa süreli kültürün 7. gününde, hücrelerin Rd123 alımında istatistiksel olarak anlamlı düşüş gözlendi p

Keywords

• Kordon kanı,kök hücre,Rodamin123,büyüme faktörleri

Determining Effects Of The Growth Factors On Stem Cell Content Of Cord Blood By Rhodamine123 Upthake Of The Cells

Abstract

Determining Effects Of The Growth Factors On Stem Cell Content Of Cord Blood By Rhodamine123 Upthake Of The Cells Objectıve: Rhodamine 123 Rh123 that accumulate in mitocondrial membran is given information about mitochondrium counts and their activities and it is used for identify for stem cells the methabolic aspects. Due to the stem cells are inactive or dormant, Rh123 accumulation is very low in these cells. In this study, we aimed to investigate the effects of growth factors on stem cell content of cord blood CB by determinig Rh123 uptake by these cells during in vitro culture period. Materıals Ve Methods: CB samples obtained from 15 healthy volunteers delivering in term. After separation, mononuclear cells incubated for 14 day presence of interleukin-3 IL-3 , IL-6, IL-11 stem cell faktör SCF , flt3/flk2 ligand FL ve trombopoetin TPO . Cells from before, day 7 and day 14 of cultures were analysed for Rh123 uptake by flow cytometry. Results: At day 7 of short term cultures initiated with growth factors known to be effective on stem cells, statistically significant p

Keywords

• Cord blood,stem cell,Rhodamin123,growth factors

References

1. Gluckman E, Rocha V. Cord blood transplant: strategy of alternative donor search. Springer Semin Immunopathol 2004; 26:143- 54. 2.
2. Grewal SS, Barker JN, Davies SM, Wagner JE. Unrelated donor haematopoietic cell transplantation: marrow or umbilical cord blood? Blood 2003; 101:4233-44. 3. Gluckman E, et Auerbach
3. reconstitution in a patient with Fanconi’s anemia by means of umbilical cord blood from an HLA-identical sibling. N Engl J Med 1989; 321:1174–8. 4. al.
4. Hematopoietic Civin CI, Strauss LC, Brovall C, Fackler MJ, Schwartz JF, Shaper JH. Antigenic analysis of hematopoiesis. III. A hematopoietic progenitor cell surface antigen defined by a monoclonal antibody raised against KG-1a cells. J Immunol. 1984; 133:157-65. 5.
5. Bhatia M, Bonnet D, Murdoch B, Gan OI, Dick JE. A newly discovered class of human hematopoietic cells with SCID- repopulating activity. Nat Med. 1998; 4:1038- 45. 6.
6. Osawa M, Hanada K, Hamada H, Nakauchi H. Long-term lymphohematopoietic reconstitution by a single CD34-low/negative hematopoietic stem cell. Science. 1996; 273:242-5. 7.
7. Goodell MA, Rosenzweig M, Kim H, et al. Dye efflux studies suggest that hematopoietic stem cells expressing low or undetectable levels of CD34 antigen exist in multiple species. Nat Med. 1997; 3:1337-45. 8.
8. Zanjani ED, Almeida-Porada G, Livingston AG, Flake AW, Ogawa M. Human bone marrow CD34- cells engraft in vivo and undergo multilineage expression that includes giving rise to CD34+ cells. Exp Hematol. 1998; 26:353-60. 9.

9. Hao QL, Shah AJ, Thiemann FT, Smogorzewska EM, Crooks GM. A functional comparison of CD34 + CD38- cells in cord blood and bone marrow. Blood. 1995; 86:3745-53. 10.
10. and activated subsets of mouse multipotent hematopoietic stem cells. Proc Natl Acad Sci U S A. 1990; 87:7433-7. 11.
11. al. Isolation of primitive human bone marrow hematopoietic progenitor cells using Hoechst 33342 and Rhodamine 123. Exp Hematol. 1996; 24:1215-24. 12.
12. Auerbach AD, Friedman HS, Douglas GW, Devergie A, Esperou H, Thierry D, Socie G, Lehn P. Hematopoietic reconstitution in a patient with Fanconi’s anemia by means of umbilical cord blood from an HLA-identical sibling. N Engl J Med 1989; 321:1174–8. 13.
13. transplant: strategy of alternative donor search. Springer Semin Immunopathol 2004; 26:143- 54. 14.
14. Wagner JE. Unrelated donor haematopoietic cell transplantation: marrow or umbilical cord blood? Blood 2003; 101:4233-44. 15.
15. Nienhuis AW, Bodine DM. Purification and haracterization of heterogeneous pluripotent hematopoietic stem cell populations expressing high levels of c-kit receptor. Blood. 1993; 82:762-70. 16.
16. Induction of c-kit molecules on human CD34+/c-kit < low cells: evidence for CD34+/c-kit < low cells as primitive hematopoietic stem cells. Stem Cells. 1997; 15:420-9. 17.
17. CD34+AC133+ cells isolated from cord blood are highly enriched in long-term culture- initiating cells, NOD/SCID-repopulating cells and dendritic cell progenitors. Stem Cells. 1998; 16:387-96. 18.
18. Alexandrunas D, Fishbaugh J, Bickenbach JR. Isolating a pure population of epidermal stem cells for use in tissue engineering. Exp Dermatol. 2001; 10:45-54 19.
19. epidermal stem cells can produce multiple cell lineages during development. Stem Cells. 2002; 20:21-31. 20.
20. The effect of cytokines on CD34+ Rh-123high and low progenitor cells from human umbilical cord blood. Exp Hematol. 1994; 22:1244-51. 21.
21. al. Does cord blood contain enough progenitor cells for transplantation? J Hematother. 1994; 3:291-8.