Abstract
BackgroundAdult bone marrow contains stem cells that replenish the myeloid and lymphoid lineages. A subset of human and mouse CD34+ bone marrow stem cells can be propagated in culture to autonomously express embryonic stem cell genes and embryonic germ layer lineage genes. The current study was undertaken to determine whether these CD34+ stem cells could be obtained from human blood, whether gene expression could be modulated by culture conditions and whether the cells produce insulin.MethodsHuman peripheral blood buffy coat cells and mobilized CD34+ cells from human blood and from blood from C57Bl/6 J mice were cultured in hybridoma medium or neural stem cell induction medium supplemented with interleukin (IL)-3, IL-6, and stem cell factor (SCF). Changes in mRNA and protein expression were assessed by Western blot analysis and by immunohistochemistry. Mass spectrometry was used to assess insulin production.ResultsWe were able to culture CD34+ cells expressing embryonic stem cell and embryonic germ layer lineage genes from adult human peripheral blood after standard mobilization procedures and from mouse peripheral blood. Gene expression could be modulated by culture conditions, and the cells produced insulin in culture.ConclusionThese results suggest a practical method for obtaining large numbers of CD34+ cells from humans to allow studies on their potential to differentiate into other cell types.
Highlights
Adult bone marrow contains stem cells that replenish the myeloid and lymphoid lineages
The current study was undertaken to determine whether CD34+ stem cells that could be induced to express embryonic germ layer lineage genes could be obtained from mobilized human peripheral blood, whether the cells so generated expressed proteins not expected in hematogenous stem cells in conditions where cell fusion was not possible, and whether gene expression could be modulated by culture conditions
The CD34+ stem cell cultures from mobilized human peripheral blood differed from those obtained from the mouse in that, while the latter contained a single spherical cell morphology, the former contained four morphological phenotypes: one cell type that was adherent to the plastic flask, and three cell types that grew in suspension—a spherical cell, a cone-shaped cell, and a minute cell
Summary
Adult bone marrow contains stem cells that replenish the myeloid and lymphoid lineages. A subset of human and mouse CD34+ bone marrow stem cells can be propagated in culture to autonomously express embryonic stem cell genes and embryonic germ layer lineage genes. The current study was undertaken to determine whether these CD34+ stem cells could be obtained from human blood, whether gene expression could be modulated by culture conditions and whether the cells produce insulin. The current study was undertaken to determine whether CD34+ stem cells that could be induced to express embryonic germ layer lineage genes could be obtained from mobilized human peripheral blood, whether the cells so generated expressed proteins not expected in hematogenous stem cells in conditions where cell fusion was not possible, and whether gene expression could be modulated by culture conditions
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a call
