Serum-Free Culture of Mid-gestation Mouse Embryos: A Tool for the Study of Endoderm-Derived Organs

2013 ◽  
pp. 183-194 ◽  
Author(s):  
Julie Gordon ◽  
Billie A. Moore ◽  
C. Clare Blackburn ◽  
Nancy R. Manley
Keyword(s):  
Mouse Embryos ◽  
Serum Free

Formulation of a complex serum-free medium (CSM) for use in the co-culture of mouse embryos with cells of the female reproductive tract

1991 ◽  
Vol 3 (1) ◽  
pp. 99 ◽  
Author(s):  
D Sakkas ◽  
AO Trounson
Keyword(s):  
Growth Factors ◽  
Growth Factor ◽  
Embryo Culture ◽  
Reproductive Tract ◽  
Culture Media ◽  
Mouse Embryos ◽  
Serum Free Medium ◽  
Free Medium ◽  
Serum Free ◽  
Cell Numbers

Co-culture of pre-implantation embryos with cells of the reproductive tract requires a medium that is beneficial to both embryos and cells. However, many studies in this area utilize media originally formulated for specific cell lines. In the present study, a complex serum-free medium (CSM) was formulated on the basis of the ionic compositions of existing embryo culture media and mouse oviductal fluid as well as the concentrations of growth factors that appear to benefit mouse embryo development. The study began by investigating the effect of altering the concentrations of K+ ions (0-40 mM) and sulfate ions (0-10 mM) in embryo culture media on the development of 2-cell mouse embryos. Mouse embryos showed improved cell numbers at the blastocyst stage when cultured in 10 mM K+ compared with Whittingham's T6 medium. Embryos were also cultured in T6 supplemented with bovine serum albumin (BSA) containing various concentrations of insulin, insulin-like growth factors I and II, fibroblast growth factor, and epidermal growth factor. Insulin concentrations of 100 ng mL-1 significantly (P less than 0.05) improved the cell numbers of 2-cell embryos cultured to the morulae and blastocyst stages compared with those cultured in T6 + BSA alone. CSM was formulated on the basis of the results of these experiments and was found to support both improved development of 2-cell mouse embryos and the culture of mouse fibroblast and mouse oviduct cells.

New serum-free in vitro culture technique for midgestation mouse embryos

genesis ◽  
2003 ◽  
Vol 35 (3) ◽  
pp. 164-168 ◽  
Author(s):  
Billie A. Moore-Scott ◽  
Julie Gordon ◽  
C. Clare Blackburn ◽  
Brian G. Condie ◽  
Nancy R. Manley
Keyword(s):  
In Vitro Culture ◽  
Culture Technique ◽  
Mouse Embryos ◽  
Serum Free

Peptides extracted from vero cell cultures overcome the blastocyst block of mouse embryos in a serum-free medium

1994 ◽  
Vol 11 (3) ◽  
pp. 165-171 ◽  
Author(s):  
Hsin-Fu Chen ◽  
Hong-Nerng Ho ◽  
Shee-Uan Chen ◽  
Kuang-Han Chao ◽  
Heng-Ru Lin ◽  
...  
Keyword(s):  
Vero Cell ◽  
Cell Cultures ◽  
Mouse Embryos ◽  
Serum Free Medium ◽  
Free Medium ◽  
Serum Free

In vitro chondrogenesis of limb mesoderm from normal and brachypod mouse embryos

Development ◽  
1975 ◽  
Vol 33 (2) ◽  
pp. 371-386
Author(s):  
William A. Elmer ◽  
David K. Selleck
Keyword(s):  
Culture Conditions ◽  
Linear Increase ◽  
Mouse Embryos ◽  
Nodule Formation ◽  
Serum Free Medium ◽  
Free Medium ◽  
Serum Free ◽  
Fresh Serum ◽  
Mitotic Figures

This paper describes the cytodifferentiation of hind limb mesodermal cells from 12-day-old normal and brachypod (bpH) mouse embryos grown in vitro at high densities. Over a 3-day culture period normal cells underwent aggregation, nodule formation, and coalescence of nodules into large masses of cartilage. This was associated at the biochemical level with a cessation of cell division, with a concomitant increase in metachromatic matrix, and synthesis of collagen. Under the described culture conditions the collagen synthesized by 48 h cultures was predominantly of cartilage type with an αl:α2 ratio of 9:1. A change in the collagen synthetic program was observed when the entire medium was replaced after 48 h incubation with fresh, serum-free medium. Under these conditionsthe αl:α2 ratio was 4:1. In contrast, brachypod cells plated at the same density appeared large, flattened, and stellate. Upon aggregation, normal nodule morphology was only rarely observed. More often large, irregular clusters formed from suspended cells loosely attaching to the surface aggregates. Concomitant with the marked changes in the morphology of the mutant cells was a linear increase in DNA synthesis and the appearance of many mitotic figures. A biochemical transformation in matrix synthesis was not observed, however. After a 24 h delay, mutant matrix accumulated and stained intensely with toluidine blue. Collagen was synthesized at approximately the normal rate and was of the cartilage type with an αl:α2 ratio of 9:1. When incubated in fresh, serum-free medium, the response of collagen subunit synthesis was identical to the normal cultures. In view of these results the possible manner in which brachypodism causes developmental anomalies of the limb skeleton is suggested.

In vitro studies on the control of trophoblast outgrowth in the mouse

Development ◽  
1973 ◽  
Vol 30 (1) ◽  
pp. 21-30
Author(s):  
E. J. Jenkinson ◽  
I. B. Wilson
Keyword(s):  
Blastocyst Stage ◽  
Mouse Embryos ◽  
In Vitro Studies ◽  
Serum Free Medium ◽  
Free Medium ◽  
Serum Free ◽  
Uterine Environment ◽  
Adhesive Interactions

When placed in serum-free medium on reconstituted collagen surfaces re-implantation mouse embryos are capable of producing characteristic trophoblast outgrowths. Previously this pattern of differentiation has been considered to be essentially dependent on the presence of serum macromolecules. Such activity is expressed only at the late blastocyst stage and is qualitatively different from the adhesive interactions between blastomeres earlier in development. The development of the properties responsible for outgrowth is intrinsic to the blastocyst, being independent of stimulation by exposure either to the uterine environment or to whole serum. The significance of these observations related to implantation control in vivo is discussed.

Exogeneous factors are not necessary in attachment, spreading and polarization of hela cells

1978 ◽  
Vol 36 (2) ◽  
pp. 310-311
Author(s):  
W. Liebrich
Keyword(s):  
Hela Cells ◽  
Calf Serum ◽  
Glass Tube ◽  
Serum Free Medium ◽  
Nacl Solution ◽  
Free Medium ◽  
Minimum Essential Medium ◽  
Serum Free ◽  
All Solutions ◽  
Glass Support

HeLa cells were grown for 2-3 days in EAGLE'S minimum essential medium with 10% calf serum (S-MEM; Seromed, München) and then incubated for 24 hours in serum free medium (MEM). After detaching the cells with a solution of 0. 14 % EDTA and 0. 07 % trypsin (Difco, 1 : 250) they were suspended in various solutions (S-MEM = control, MEM, buffered salt solutions with or without Me++ions, 0. 9 % NaCl solution) and allowed to settle on glass tube slips (Leighton-tubes). After 5, 10, 15, 20, 25, 30, 1 45, 60 minutes 2, 3, 4, 5 hours cells were prepared for scanning electron microscopy as described by Paweletz and Schroeter. The preparations were examined in a Jeol SEM (JSM-U3) at 25 KV without tilting.The suspended spherical HeLa cells are able to adhere to the glass support in all solutions. The rate of attachment, however, is faster in solutions without serum than in the control. The latter is in agreement with the findings of other authors.

Human Adenovirus Uptake by Preirnplantation Mouse Embryos

1972 ◽  
Vol 30 ◽  
pp. 268-269
Author(s):  
D. G. Chase ◽  
W. Winters ◽  
L. Piko
Keyword(s):  
Hela Cells ◽  
Human Adenovirus ◽  
Mouse Embryos ◽  
Equilibrium Density ◽  
Cell Stage ◽  
Virus Attachment ◽  
Preimplantation Mouse Embryos ◽  
Embryo Culture Medium ◽  
Preimplantation Mouse ◽  
Mouse Cells

Although the outlines of human adenovirus entry and uncoating in HeLa cells has been clarified in recent electron microscope studies, several details remain unclear or controversial. Furthermore, morphological features of early interactions of human adenovirus with non-permissive mouse cells have not been extensively documented. In the course of studies on the effects of human adenoviruses type 5 (AD-5) and type 12 on cultured preimplantation mouse embryos we have examined virus attachment, entry and uncoating. Here we present the ultrastructural findings for AD-5.AD-5 was grown in HeLa cells and purified by successive velocity gradient and equilibrium density gradient centrifugations in CsCl. After dialysis against PBS, virus was sedimented and resuspended in embryo culture medium. Embryos were placed in culture at the 2-cell stage in Brinster's medium.

Preplate neurons in embryonic mouse cortex: Ultrastructural observations using photoconversion of the fluorescent lipophilic dye dil

1992 ◽  
Vol 50 (1) ◽  
pp. 906-907
Author(s):  
Linda C. Hassinger ◽  
James E. Crandall
Keyword(s):  
Mouse Embryos ◽  
Embryonic Mouse ◽  
Cortical Afferents ◽  
Mouse Cortex ◽  
Carbocyanine Dye ◽  
Increased Sensitivity

We have begun to look directly at small numbers of afferent axons to early generated neurons that form the preplate in the developing mouse cortex. The carbocyanine dye Dil (1’1, dioctadecyl-3,3,3’3’-tetramethyl-indocarbocyanine) has proved especially useful for this goal. DiI labels axons and their terminals with greater sensitivity and without some of the disadvantages of axon filling with HRP. The increased sensitivity provided by labeling embryonic axons with DiI has given us new insights into the development of cortical afferents. For instance, we reported originally that afferents from the thalamus were present below the cortex as early as embryonic day 15 (E15) based on HRP injections into mouse embryos. By using DiI placements into the thalamus in aldehyde-fixed brains, we now know that thalamic fibers reach the cortex 24 hrs earlier.

Localization of Intracisternal a Particle Antigens in Neoplastic Cells and Preimplantation Mouse Embryos

1980 ◽  
Vol 38 ◽  
pp. 696-697
Author(s):  
Thomas T.F. Huang ◽  
Patricia G. Calarco
Keyword(s):  
Endoplasmic Reticulum ◽  
Normal Development ◽  
Mouse Embryos ◽  
Neoplastic Cells ◽  
Large Numbers ◽  
Preimplantation Mouse Embryos ◽  
Preimplantation Mouse ◽  
Intracisternal A Particle ◽  
Normal Feature

The stage specific appearance of a retravirus, termed the Intracisternal A particle (IAP) is a normal feature of early preimplantation development. To date, all feral and laboratory strains of Mus musculus and even Asian species such as Mus cervicolor and Mus pahari express the particles during the 2-8 cell stages. IAP form by budding into the endoplasmic reticulum and appear singly or as groups of donut-shaped particles within the cisternae (fig. 1). IAP are also produced in large numbers in several neoplastic cells such as certain plasmacytomas and rhabdomyosarcomas. The role of IAP, either in normal development or in neoplastic behavior, is unknown.

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