CSF-1 and mouse preimplantation development in vitro

Development ◽  
1995 ◽  
Vol 121 (5) ◽  
pp. 1333-1339 ◽  
Author(s):  
P. Bhatnagar ◽  
V.E. Papaioannou ◽  
J.D. Biggers

The effects of macrophage colony stimulating factor on the development of the zygote to the blastocyst stage of an outbred strain of mouse have been studied in KSOM, an improved medium that supports a high rate of in vitro development. Macrophage colony stimulating factor accelerates the formation of the blastocyst cavity by day 4 (96 hours post-hCG). It also increases overall embryonic cell number through a differential increase in the number of trophoblast cells, with no significant effect on the number of inner cell mass cells. By day 5 of culture (120 hours post-hCG), colony stimulating factor-treated embryos have about 20 more trophoblast cells than control embryos, an increase of about 30 percent of the total number of cells in a control blastocyst. The maximum response of embryos was obtained at a concentration around 540 U ml-1 colony stimulating factor (identical to 918 Stanley units ml-1), and the cytokine can produce the same effects even if it is present in the medium for only part of the culture period. This in vitro stimulation of preimplantation development with macrophage colony stimulating factor is compatible with continued normal fetal development in vivo.

2006 ◽  
Vol 18 (2) ◽  
pp. 191
Author(s):  
D. H. Kim ◽  
S. W. Kim ◽  
B. C. Yang ◽  
G. S. Im ◽  
H. S. Park ◽  
...  

Granulocyte-macrophage colony stimulating factor (GM-CSF) is secreted by epithelial cells lining the female reproductive tract in mice and several other species. GM-CSF receptors are present in the fertilized oocyte and in all subsequent stages of development, and in blastocysts it is expressed in both inner cell mass and trophectoderm cells. Recent studies suggest that GM-CSF can act as a survival factor for the developing embryo. The purpose of this study was to examine the effect of GM-CSF, as a medium supplement, on the development of nuclear-transferred bovine embryos. Oocytes were enucleated after in vitro maturation in TCM-199 supplemented with 10% fetal bovine serum (FBS), 1 mg/mL FSH, and 1 mg/mL estradiol-17� for 20 h. Enucleated oocytes were fused with bovine ear skin fibroblast cells by a DC pulse of 25 V/150 mm for 20 ms in Zimmerman cell fusion medium. For activation, reconstructed embryos were exposed to 10 mM Ca-ionophore for 5 min, followed by 2 mM 6-dimethylaminopurine for 3 h. NT embryos were subsequently cultured in CR2 medium without or with 10 ng/mL recombinant porcine GM-CSF at 39.0�C in 5% O2, 5% CO2 and 90% N2. After 7 days of culture, blastocyst formation was observed. The number of inner cell mass (ICM) and trophectoderm (TE) cells was examined by differential staining. Apoptotic cells in blastocysts were detected by a terminal deoxynucleotidyl transferase-mediated d-UTP nick-end labeling (TUNEL) assay. Data were analyzed by chi-square and Student's t-test. Addition of GM-CSF to the medium significantly (P < 0.05) increased the proportion of embryos developing to the blastocyst stage (37.6 � 12.0 and 54.7 � 13.9% for control and GM-CSF groups respectively). No differences in the total cell number and the ratio of ICM to total cells were detected between the control group (125.4 � 35.7 and 38.5 � 9.7%) and the GM-CSF group (123.8 � 35.1 and 34.2 � 13.1%). The mean proportion of apoptotic cells in blastocysts was not different between the control (5.4 � 5.4%) and the GM-CSF (5.3 � 3.9%) group. Our results showed the beneficial effect of GM-CSF on the development of NT bovine embryos. These results suggest that GM-CSF might be a useful molecule for increasing development of NT bovine embryos. Further studies are necessary to verify the mechanism of GM-CSF on the development of bovine NT embryos.


2000 ◽  
Vol 204 (2) ◽  
pp. 114-127 ◽  
Author(s):  
Roberto P. Revoltella ◽  
Leopoldo Laricchia Robbio ◽  
Anna Marina Liberati ◽  
Gigliola Reato ◽  
Robin Foa ◽  
...  

Blood ◽  
1988 ◽  
Vol 72 (4) ◽  
pp. 1329-1332 ◽  
Author(s):  
DC Kaufman ◽  
MR Baer ◽  
XZ Gao ◽  
ZQ Wang ◽  
HD Preisler

Expression of the granulocyte-macrophage colony-stimulating factor (GM- CSF) gene in acute myelocytic leukemia (AML) was assayed by Northern blot analysis. GM-CSF messenger RNA (mRNA) was detected in the freshly obtained mononuclear cells of only one of 48 cases of AML, in contrast with recent reports that GM-CSF mRNA might be detected in half of the cases of AML when RNA is prepared from T-cell- and monocyte-depleted leukemic cells. We did find, however, that expression of the GM-CSF gene was detectable in five of ten cases after in vitro T-cell and monocyte depletion steps. Additional studies suggest that expression of GM-CSF in the bone marrow of the one positive case, rather than being autonomous, was under exogenous control, possibly by a paracrine factor secreted by marrow stromal cells. These studies emphasize the potential for altering in vivo patterns of gene expression by in vitro cell manipulation.


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