Measurement of hemoglobin in long-term fixed erythroid cells: application development for cell science experiments in microgravity.
Studying the effects of microgravity on cell differentiation will enhance our understanding of fundamental biology and is indispensable for a sustained space program. Rauscher murine erythroleukemic cells were chosen as a model system to study erythroid cell differentiation aboard the International Space Station because these cells undergo differentiation in response to the natural inducer, erythropoietin, as well as various chemical-inducers. We have now developed a method to quantify hemoglobin in Rauscher cells after weeks of fixation and storage required for such space biology experiments. By exploiting the pseudoperoxidase activity of hemoglobin and by using reagents that yield a soluble chromophore that freely passes out of fixed cells, we developed a highly specific and sensitive assay applicable to cells fixed as long as 4 months.
Xu
Kun
K
Laboratory for Cellular and Molecular Biology, Division of Hematology and Oncology, Beth Israel Deaconess Medical Center, Department of Medicine, Harvard Medical School, Boston, Massachusetts 02215, USA.
Davis
Kerry L
KL
Sytkowski
Arthur J
AJ
D000818:Animals; D001560:Benzidines / Q000737:chemistry; D002478:Cells, Cultured; D041905:Erythroid Cells / Q000166:cytology / Q000378:metabolism*; D004920:Erythropoiesis / Q000502:physiology*; D004921:Erythropoietin; D005404:Fixatives / Q000737:chemistry; D006454:Hemoglobins / Q000378:metabolism*; D006861:Hydrogen Peroxide / Q000737:chemistry; D051379:Mice; D009195:Peroxidase / Q000378:metabolism; D013026:Space Flight; D014893:Weightlessness
D001560:Benzidines; D005404:Fixatives; D006454:Hemoglobins; D004921:Erythropoietin; C021758:3,3',5,5'-tetramethylbenzidine; D006861:Hydrogen Peroxide; D009195:Peroxidase