Turning brain into blood: A hematopoietic fate adopted by adult neural stem cells in vivo

Science

Volumn 283, Issue 5401, 1999, Pages 534-537

  Bjornson, Christopher R R ^a   Rietze, Rodney L ^a   Reynolds, Brent A ^a   Magli, M Cristina ^a   Vescovi, Angelo L ^a  

^a NONE

Author keywords

[No Author keywords available]

Indexed keywords

ARTICLE; BONE MARROW CELL; CELL DIFFERENTIATION; CELL POPULATION; CELL TYPE; HEMATOPOIETIC STEM CELL; IRRADIATION; LYMPHOID CELL; OLIGODENDROGLIA; PRIORITY JOURNAL;

ANIMALS; BLOOD CELLS; BONE MARROW CELLS; CELL DIFFERENTIATION; CELLS, CULTURED; COLONY-FORMING UNITS ASSAY; FEMALE; H-2 ANTIGENS; HEMATOPOIESIS; HEMATOPOIETIC STEM CELLS; LAC OPERON; LYMPHOCYTES; MALE; MICE; MICE, INBRED BALB C; MICE, TRANSGENIC; PROSENCEPHALON; SPLEEN; STEM CELL TRANSPLANTATION; STEM CELLS;

EID: 0033593654     PISSN: 00368075     EISSN: None     Source Type: Journal    
DOI: 10.1126/science.283.5401.534     Document Type: Article

References (25)

1. M. Loeffler and C. S. Potten, in Stem Cells, C. S. Potten, Ed. (Academic Press, Cambridge, MA, 1997), pp. 1-27.
2. B. A. Reynolds and S. Weiss, Science 255, 1707 (1992); A. Gritti et al., J. Neurosci. 16, 1091 (1996).
3. B. A. Reynolds and S. Weiss, Science 255, 1707 (1992); A. Gritti et al., J. Neurosci. 16, 1091 (1996).
4. S. J. Morrison, N. M. Shah, D. J. Anderson, Cell 88, 287 (1997); R. D. G. McKay Science 276, 66 (1997).
5. S. J. Morrison, N. M. Shah, D. J. Anderson, Cell 88, 287 (1997); R. D. G. McKay Science 276, 66 (1997).
6. S. Tajbakhsh et al. Neuron 13, 813 (1994); N. L. M. Valtz, T. E. Hayes, T. Norregaard, S. Liu, R. D. G. McKay, New Biol 3, 364 (1991).
7. S. Tajbakhsh et al. Neuron 13, 813 (1994); N. L. M. Valtz, T. E. Hayes, T. Norregaard, S. Liu, R. D. G. McKay, New Biol 3, 364 (1991).
8. N. J. Tarbell et al. Int. J. Radiat. Oncol. Biol. Phys. 13, 1065 (1987).
9. NSCs were isolated from 14-day-old embryonic and adult striata of ROSA26 animals (Jackson Laboratories) as previously described (3). Cultures reported here were of early passage number (12 to 20), although cells of later passage (30 to 35) produced identical results. Cultures were harvested 6 days after the last passage. The growth media used was as previously described (2) with epidermal growth factor(20 ng/ml) and fibroblast growth factor 2 (10 ng/ml) as mitogens.
10. 7 ROSA26 BM in 200 μl of Earle's buffered saline solution (EBSS; Gibco-BRL) through the tail vein 16 hours after irradiation (n = 4 independent experiments). Alt animal experimentation protocols were approved by the University of Calgary Animal Care Services.
11. G. Friedrich and P. Sorriano, Genes Dev. 5, 1513 (1991).
12. P. Bartlett, Proc. Natl. Acad. Sci. U.S.A. 79, 2722 (1982); F. Alliot, E. Lacain, B. Grima, B. Pessac, ibid. 88, 1541 (1991).
13. P. Bartlett, Proc. Natl. Acad. Sci. U.S.A. 79, 2722 (1982); F. Alliot, E. Lacain, B. Grima, B. Pessac, ibid. 88, 1541 (1991).
14. The primer dropping method [H Wong, W. D. Anderson, T. Cheng, K. T. Riabowol, Anal. Biochem. 223, 251 (1994)] was used to amplify genomic DNA by PCR. Forty cycles of 94° (1 min), 60°C (1 min), and 72°C (1 min) were used to amplify lacZ with the following primer pair: 5′-TTG GAG TGA CGC CAG TTA TCT GGA and 3′-TCA ACC ACC GCA CGA TAG AGA TTC. After 20 cycles, primers specific for glyceraldehyde-3-phosphate dehydrogenase (CAPDH; 5′-CGG AGT CAA CCG ATT TGG TCG TAT and 3′-AGC CTT CTC CAT GGT GGT GAA GAC) were added as an internal control.
15. 2 atmosphere. Cytokines used were the following: interleukin-3 (IL-3) (10 ng/ml), IL-7 (10 ng/ml), stem cell factor (50 ng/ml), erythropoietin (3 U/ml; R&D Systems), and IL-6 (10 ng/ml; Novartis).
16. 2 (Sigma)] and X-Gal (5-bromo-4-chloro-3-indoxyl-β-D-galactopyranoside; 1 mg/ml; in dimethyl sulfoxide; Molecular Probes) in phosphate-buffered saline (PBS) (pH 7.4) were added to methylcellulose cultures for 8 hours at 37°C.
17. 6 cells were added to 100 μl of FACS buffer supplemented with the appropriate primary antibodies and incubated at 4°C for 30 min. After washing, secondary antibodies were added (whete appropriate) and incubated at 4°C for 30 min. For biotinylated antibodies, isotype controls were used to set gates; otherwise, gates were set with cells alone. Cell viability was greater than 95%, by propidium iodide exclusion. Flow cytometric analysis was performed with a FACScan (Becton-Dickinson) with all events gated on the forward and side scatter.
18. d) (PharMingen) were used.
19. Clones 145-2c11 (CD3e), 1D3 (CD19), and M1/70 (CD11b) (PharMingen) were used.
20. M. A. Di Berardino, Genomic Potential of Differentiated Cells (Columbia Univ. Press, New York, 1997), pp. 1-3.
21. I. Wilmut, A. E. Schnieke, J. McWhir, A. J. Kind, K. H. S. Campbell, Nature 385, 810 (1997).
22. A. L Vescovi et al., Exp. Neurol., in press; J. D. Flax et al., Nature Biotechnot. 16, 1033 (1998).
23. A. L Vescovi et al., Exp. Neurol., in press; J. D. Flax et al., Nature Biotechnot. 16, 1033 (1998).
24. Single- and triple-label immunocytochemistry was performed as previously described (2) with antibody to nestin, monoclonal antibody to type III β-tubulin (Sigma), glial fibrillary acidic protein antisera (Incstar), and monoclonal antibody to O4 (immunoglobulin M; Boehringer Mannheim).
25. We thank M. Dicay, L. Robertson, D. Schmidt, G. Gobbel, D. Spencer, R. Dawson, and P. Hill for technical assistance; E. Cattaneo and R. McKay for antibody to nestin; and J. Dick for critical assessment of this work. Supported by the Spinal Cord Society, Fergus Falls, MN, and by Comitato Telethon (grant A. 116) to A.L.V.