Separation of Spermatogenic Cells From Adult Transgenic Mouse Testes Using Unit-Gravity Sedimentation

2003 ◽  
Vol 25 (2) ◽  
pp. 131-138 ◽  
Author(s):  
Susan M. Wykes ◽  
Stephen A. Krawetz
Keyword(s):  
Transgenic Mouse ◽  
Spermatogenic Cells ◽  
Gravity Sedimentation ◽  
Unit Gravity Sedimentation

scholarly journals Separation of Erythropoietin-sensitive Cells From Hemopoietic Spleen Colony-forming Stem Cells of Mouse Fetal Liver by Unit Gravity Sedimentation

Blood ◽  
1971 ◽  
Vol 37 (4) ◽  
pp. 417-427 ◽  
Author(s):  
JOHN R. STEPHENSON ◽  
ARTHUR A. AXELRAD
Keyword(s):  
Stem Cells ◽  
Present Method ◽  
Fetal Liver ◽  
Sedimentation Velocity ◽  
Assay Method ◽  
Fetal Liver Cells ◽  
Gravity Sedimentation ◽  
Cell Fractions ◽  
Unit Gravity Sedimentation

Abstract An assay method for erythropoietin-sensitive cells has been developed based on the fact that cells of mouse fetal liver respond to erythropoietin in vitro by increased heme synthesis. To determine whether or not hemopoietic colony-forming stem cells are identical with erythropoietin-sensitive cells, C3H/Bi 13-day fetal liver cells were separated into fractions on the basis of size by unit gravity sedimentation through a 1-2 per cent bovine serum albumin gradient. The cell fractions obtained were assayed for erythropoietin-sensitive cells by the present method and for spleen colony-forming cells by the method of Till and McCulloch. It was found that the modal sedimentation velocity of erythropoietin-sensitive cells was greater than that of the spleen colony-forming cells of mouse fetal liver, showing that these two classes of cells are distinct.

scholarly journals Generation of flagella by cultured mouse spermatids.

1984 ◽  
Vol 98 (2) ◽  
pp. 619-628 ◽  
Author(s):  
G L Gerton ◽  
C F Millette
Keyword(s):  
Electron Microscopy ◽  
Spermatogenic Cells ◽  
Fibrous Sheath ◽  
Acridine Orange Staining ◽  
Transmission Electron ◽  
Fetal Bovine ◽  
Outer Dense Fibers ◽  
Unit Gravity Sedimentation

During the short-term culturing of mouse spermatogenic cells, flagella were generated by round spermatids previously lacking tails. Unseparated germ cells were obtained by enzymatic treatments and round spermatids (greater than 90% pure) were purified by unit gravity sedimentation. As determined by Nomarski or phase-contrast microscopy, no cells had flagella immediately after isolation; flagella were first clearly detected after 6 1/2 h of culture in Eagle's minimal essential medium containing 10% fetal bovine serum and 6 mM lactate. After 24 h, approximately 20% of round spermatids had formed flagella. Multinucleated round spermatids often formed multiple flagella, the number never exceeding the number of nuclei per symplast. Round spermatids were the only spermatogenic cells capable of tail formation. Flagella elongation was blocked by 1 microM demecolcine, an inhibitor of tubulin polymerization. Indirect immunofluorescence localized tubulin in the flagella. As seen by scanning electron microscopy, flagella developed as early as 2 h after culture and continued to elongate over the next 20 h, reaching lengths of at least 19 micron. Transmission electron microscopy demonstrated that flagella formed in culture resembled flagella from Golgi-phase round spermatids in situ; the flagella consisted of "9+2" axonemes lacking other accessory structures such as outer dense fibers and the fibrous sheath. As determined by acridine orange staining of the developing acrosomes, all spermatids that formed flagella in culture were Golgi-phase spermatids. By these criteria, the structures are indeed true flagella, corresponding in appearance to what others have described for early mammalian spermatid flagella in situ. We believe this is the first substantiated report of limited in vitro differentiation by isolated mammalian spermatids.

An improved method of unit gravity sedimentation separation for murine peritoneal macrophage populations

1983 ◽  
Vol 61 (1) ◽  
pp. 117-123 ◽  
Author(s):  
R.A. De Weger ◽  
J.M.A. Verbakel ◽  
R. Oskam ◽  
H.T.C.M. Van Den Berg ◽  
C.D.H. Van Basten ◽  
...  
Keyword(s):  
Peritoneal Macrophage ◽  
Improved Method ◽  
Murine Peritoneal Macrophage ◽  
Gravity Sedimentation ◽  
Macrophage Populations ◽  
Unit Gravity Sedimentation

A new unit gravity sedimentation chamber

1980 ◽  
Vol 105 (1) ◽  
pp. 246-256 ◽  
Author(s):  
A. Tulp ◽  
J.G. Collard ◽  
A.A.M. Hart ◽  
J.A. Aten
Keyword(s):  
Gravity Sedimentation ◽  
Unit Gravity Sedimentation

Isolation of Hybridoma Forming Cells From Immune Spleens by Unit Gravity Sedimentation

Hybridoma ◽  
1991 ◽  
Vol 10 (4) ◽  
pp. 529-538 ◽  
Author(s):  
KRISTINE KUUS-REICHEL ◽  
AMY BEEBE ◽  
CHRISTINE KNOTT
Keyword(s):  
Gravity Sedimentation ◽  
Unit Gravity Sedimentation

Action of thyrotropin-releasing hormone on mammotrophs and thyrotrophs

1981 ◽  
Vol 241 (4) ◽  
pp. E298-E304
Author(s):  
G. Snyder ◽  
Z. Naor ◽  
C. P. Fawcett ◽  
S. M. McCann
Keyword(s):  
Anterior Pituitary ◽  
Thyrotropin Releasing Hormone ◽  
Female Rats ◽  
Pituitary Cells ◽  
Cell Distribution ◽  
Camp Content ◽  
Releasing Hormone ◽  
Gravity Sedimentation ◽  
Unit Gravity Sedimentation ◽  
Region Ii

Anterior pituitary cells from 15-day female rats were separated by unit gravity sedimentation into four populations (designated regions I-IV) based on the profile of cell distribution and the resulting content of radioimmunoassayable (RIA) hormones. The cells in regions II and IV released thyrotropin (TSH) in response to thyrotropin-releasing hormone (TRH, 5 ng/ml); however, those in region IV released only approximately 5% of their RIA content, whereas those in region II released approximately 26% in response to the same stimulus. Concomitant elevation of cAMP and of cGMP occurred in region II cells but only cGMP was elevated in region IV cells. Mammotrophs were localized in region III. They responded to TRH by releasing prolactin (PRL) and exhibiting increased cAMP content. These data provide support for the existence of two functionally distinct populations of thyrotrophs in 15-day-old female rats. The data also imply that cAMP is involved in TRH induced PRL release, whereas cGMP is involved in TRH-induced TSH release.

Haemoglobin and globin synthesis in the isolated primitive and definitive erythroid cells of chicken embryos. Evidence for a non-clonal mechanism at the haemoglobin switch

Development ◽  
1984 ◽  
Vol 84 (1) ◽  
pp. 125-148
Author(s):  
Margreet Schalekamp ◽  
Diety van Goor
Keyword(s):  
Gel Electrophoresis ◽  
Cell Types ◽  
Starch Gel Electrophoresis ◽  
Erythroid Cells ◽  
Isoelectric Focussing ◽  
Chicken Embryos ◽  
Starch Gel ◽  
Gravity Sedimentation ◽  
Globin Synthesis ◽  
Unit Gravity Sedimentation

Primitive and definitive erythroid cells of chicken embryos aged 4–8 days, were separated by unit gravity sedimentation and pulse labelled with [3H]- and [14C]leucine. The haemoglobin and globin synthesis in the cell populations was analysed by chromatofocussing, isoelectric focussing, urea starch gel electrophoresis, and immunofluorescence or radioimmunoassay, using globin specific antibodies. We found that both embryonic and adult α globins are present in primitive erythroid cells, but relatively more of the adult α-type globins are synthesized in the late primitive erythroid cells. In young definitive erythroid cells exclusively adult α-type globins are synthesized. From these findings we conclude that a command to synthesize adult αglobin is perceived in both cell types at the time of the switch. This supports an environmental model rather than a clonal model of haemoglobin switching.

scholarly journals Fractionation of human spermatogenic cells using STA-PUT gravity sedimentation and their miRNA profiling

2015 ◽  
Vol 5 (1) ◽  
Author(s):  
Yun Liu ◽  
Minghui Niu ◽  
Chencheng Yao ◽  
Yanan Hai ◽  
Qingqing Yuan ◽  
...  
Keyword(s):  
Spermatogenic Cells ◽  
Mirna Profiling ◽  
Gravity Sedimentation
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