Your browser doesn't support javascript.
loading
Immunophenotype of hematopoietic stem cells from placental/umbilical cord blood after culture
Pranke, P; Hendrikx, J; Debnath, G; Alespeiti, G; Rubinstein, P; Nardi, N; Visser, J.
  • Pranke, P; Universidade Federal do Rio Grande do Sul. Faculdade de Farmácia. Laboratório de Hematologia. Porto Alegre. BR
  • Hendrikx, J; New York Blood Center. Stem Cell Biology. New York. US
  • Debnath, G; New York Blood Center. Stem Cell Biology. New York. US
  • Alespeiti, G; New York Blood Center. Stem Cell Biology. New York. US
  • Rubinstein, P; New York Blood Center. Immunogenetics. New York. US
  • Nardi, N; Universidade Federal do Rio Grande do Sul. Departamento de Genética. Porto Alegre. BR
  • Visser, J; New York Blood Center. Stem Cell Biology. New York. US
Braz. j. med. biol. res ; 38(12): 1775-1789, Dec. 2005.
Article in English | LILACS | ID: lil-417200
ABSTRACT
Identification and enumeration of human hematopoietic stem cells remain problematic, since in vitro and in vivo stem cell assays have different outcomes. We determined if the altered expression of adhesion molecules during stem cell expansion could be a reason for the discrepancy. CD34+CD38- and CD34+CD38+ cells from umbilical cord blood were analyzed before and after culture with thrombopoietin (TPO), FLT-3 ligand (FL) and kit ligand (KL; or stem cell factor) in different combinations TPO + FL + KL, TPO + FL and TPO, at concentrations of 50 ng/mL each. Cells were immunophenotyped by four-color fluorescence using antibodies against CD11c, CD31, CD49e, CD61, CD62L, CD117, and HLA-DR. Low-density cord blood contained 1.4 ± 0.9 percent CD34+ cells, 2.6 ± 2.1 percent of which were CD38-negative. CD34+ cells were isolated using immuno-magnetic beads and cultured for up to 7 days. The TPO + FL + KL combination presented the best condition for maintenance of stem cells. The total cell number increased 4.3 ± 1.8-fold, but the number of viable CD34+ cells decreased by 46 ± 25 percent. On the other hand, the fraction of CD34+CD38- cells became 52.0 ± 29 percent of all CD34+ cells. The absolute number of CD34+CD38- cells was expanded on average 15 ± 12-fold when CD34+ cells were cultured with TPO + FL + KL for 7 days. The expression of CD62L, HLA-DR and CD117 was modulated after culture, particularly with TPO + FL + KL, explaining differences between the adhesion and engraftment of primary and cultured candidate stem cells. We conclude that culture of CD34+ cells with TPO + FL + KL results in a significant increase in the number of candidate stem cells with the CD34+CD38- phenotype.
Subject(s)
Full text: Available Index: LILACS (Americas) Main subject: Hematopoietic Stem Cells / Immunophenotyping / Fetal Blood Type of study: Prognostic study Limits: Humans / Infant, Newborn Language: English Journal: Braz. j. med. biol. res Journal subject: Biology / Medicine Year: 2005 Type: Article Affiliation country: Brazil / United States Institution/Affiliation country: New York Blood Center/US / Universidade Federal do Rio Grande do Sul/BR

Similar

MEDLINE

...
LILACS

LIS

Full text: Available Index: LILACS (Americas) Main subject: Hematopoietic Stem Cells / Immunophenotyping / Fetal Blood Type of study: Prognostic study Limits: Humans / Infant, Newborn Language: English Journal: Braz. j. med. biol. res Journal subject: Biology / Medicine Year: 2005 Type: Article Affiliation country: Brazil / United States Institution/Affiliation country: New York Blood Center/US / Universidade Federal do Rio Grande do Sul/BR