Progenitor/stem cell fate determination: interactive dynamics of cell cycle and microvesicles.
We have shown that hematopoietic stem/progenitor cell phenotype and differentiative potential change throughout cell cycle. Lung-derived microvesicles (LDMVs) also change marrow cell phenotype by inducing them to express pulmonary epithelial cell-specific mRNA and protein. These changes are accentuated when microvesicles isolated from injured lung. We wish to determine if microvesicle-treated stem/progenitor cell phenotype is linked to cell cycle and to the injury status of the lung providing microvesicles. Lineage depleted, Sca-1+ (Lin-/Sca-1+) marrow isolated from mice were cultured with interleukin 3 (IL-3), IL-6, IL-11, and stem cell factor (cytokine-cultured cells), removed at hours zero (cell cycle phase G0/G1), 24 (late G1/early S), and 48 (late S/early G2/M), and cocultured with lung tissue, lung conditioned media (LCM), or LDMV from irradiated or nonirradiated mice. Alternatively, Lin-/Sca-1+ cells not exposed to exogenous cytokines were separated into G0/G1 and S/G2/M cell cycle phase populations by fluorescence-activated cell sorting (FACS) and used in coculture. Separately, LDMV from irradiated and nonirradiated mice were analyzed for the presence of adhesion proteins. Peak pulmonary epithelial cell-specific mRNA expression was seen in G0/G1 cytokine-cultured cells cocultured with irradiated lung and in late G1/early S cells cocultured with nonirradiated lung. The same pattern was seen in cytokine-cultured Lin-/Sca-1 cells cocultured with LCM and LDMV and when FACS-separated Lin-/Sca-1 cells unexposed to exogenous cytokines were used in coculture. Cells and LDMV expressed adhesion proteins whose levels differed based on cycle status (cells) or radiation injury (LDMV), suggesting a mechanism for microvesicle entry. These data demonstrate that microvesicle modification of progenitor/stem cells is influenced by cell cycle and the treatment of the originator lung tissue.
Division of Hematology and Oncology, Rhode Island Hospital, The Warren Alpert Medical School of Brown University, Providence, RI 02903, USA. jason_aliotta@brown.edu
Jason M
JM
Aliotta
David
D
Lee
Napoleon
N
Puente
Sam
S
Faradyan
Edmund H
EH
Sears
Ashley
A
Amaral
Laura
L
Goldberg
Mark S
MS
Dooner
Mandy
M
Pereira
Peter J
PJ
Quesenberry
D000818:Animals; D001854:Bone Marrow Cells / Q000378:metabolism / Q000502:physiology*; D015815:Cell Adhesion Molecules / Q000235:genetics / Q000378:metabolism; D002453:Cell Cycle; D002454:Cell Differentiation; D002478:Cells, Cultured; D018920:Coculture Techniques; D017077:Culture Media, Conditioned; D016207:Cytokines / Q000502:physiology; D020869:Gene Expression Profiling; D008168:Lung / Q000166:cytology / Q000528:radiation effects; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D037661:Pulmonary Surfactant-Associated Proteins / Q000378:metabolism; D013234:Stem Cells / Q000378:metabolism / Q000502:physiology*; D022161:Transport Vesicles / Q000378:metabolism / Q000502:physiology*
D015815:Cell Adhesion Molecules; D017077:Culture Media, Conditioned; D016207:Cytokines; D037661:Pulmonary Surfactant-Associated Proteins