Development of embryonic rat eyes in organ culture

Development ◽  
1968 ◽  
Vol 19 (3) ◽  
pp. 397-405
Author(s):  
R. Christy Armstrong ◽  
Joel J. Elias

Previous studies on the differentiation of whole embryonic eyes in culture have been carried out mainly in the chick (Strangeways & Fell, 1926; Dorris, 1938; Harrison & Berry, 1959) and in one case in the rat (Tansley, 1933). Early postnatal intact eyes of rats and mice were cultured by Lucas & Trowell (1958), while more recently intact and trypsinized early eye rudiments (9 and 10 days) of mouse embryos have been grown in various modified organ culture systems (Muthukkaruppan, 1965). In all these experiments the media employed were either entirely biological or chemically defined media supplemented with biological media. Also, with the exception of the work done by Muthukkaruppan (1965), eye differentiation was well advanced at the start of the culture period. The present study was designed to test the feasibility of observing early embryonic rat eye development in organ culture with a totally defined medium.

1972 ◽  
Vol 18 (7) ◽  
pp. 1087-1090 ◽  
Author(s):  
C. P. Kenny ◽  
B. B. Diena ◽  
R. Wallace ◽  
L. Greenberg

Neisseria Chemically Defined Medium (NCDM) has been used routinely in our laboratory for a variety of purposes. The present report describes the development of NCDM agar, wherein the NCDM base is sterilized by filtration and defined supplements and agar are added. The medium is transparent and both meningococci and gonococci grow within 72 h. When grown on NCDM agar, Types 2 and 3 gonococcal colonies tend to revert to Type 1. The serological grouping of meningococci with specific antisera is not affected by growth on this medium.Parallel investigations on the growth of these species in liquid NCDM demonstrated that the yield of Neisseria gonorrhoeae is enhanced when the medium is sterilized by filtration.


2016 ◽  
Vol 26 (4) ◽  
pp. 269-276
Author(s):  
Jinfeng Shao ◽  
Marcelo F.M. Marcondes ◽  
Vitor Oliveira ◽  
Jaap Broos

Chemically defined media for growth of <i>Lactococcus lactis</i> strains contain about 50 components, making them laborious and expensive growth media. However, they are crucial for metabolism studies as well as for expression of heterologous proteins labeled with unnatural amino acids. In particular, the <i>L. lactis</i> Trp auxotroph PA1002, overexpressing the tryptophanyl tRNA synthetase enzyme of <i>L. lactis</i>, is very suitable for the biosynthetic incorporation of Trp analogs in proteins because of its most relaxed substrate specificity reported towards Trp analogs. Here we present two much simpler defined media for <i>L. lactis</i>, which consist of only 24 or 31 components, respectively, and with which the <i>L. lactis</i> Trp auxotroph shows similar growth characteristics as with a 50-component chemically defined medium. Importantly, the expression levels of two recombinant proteins used for evaluation were up to 2-3 times higher in these new media than in the 50-component medium, without affecting the Trp analog incorporation efficiency. Taken together, the simplest chemically defined media reported so far for <i>L. lactis</i> are presented. Since<i> L. lactis</i> also shows auxotrophy for Arg, His, Ile, Leu Val, and Met, our simplified media may also be useful for the biosynthetic incorporation of analogs of these five amino acids.


2008 ◽  
Vol 60 (4) ◽  
pp. 509-517 ◽  
Author(s):  
Jelena Lozo ◽  
Jelena Begovic ◽  
B. Jovcic ◽  
Natasa Golic ◽  
L. Topisirovic

The purpose of this study was to determine the ability of natural isolates of lactobacilli from different ecological niches to grow in a chemically defined medium in the presence or absence of sulphur-containing amino acids, methionine and/or cysteine. The obtained results indicate that cysteine is essential for growth of L. paracasei subsp. paracasei BGHN14 and BGSJ2-8, while methionine is essential for isolates BGHN40, BGCG31, and BGHV54T of the species L. plantarum. Methionine is also essential for growth of L. rhamnosus BGHV58T. Other analyzed strains, such as L. plantarum BGSJ3-18, BGZB19, BGHV52Ta, and BGHV43T, require the presence of both amino acids for their growth.


1996 ◽  
Vol 70 (2) ◽  
pp. 173-175 ◽  
Author(s):  
H. Hata

AbstractThird-stage larvae of Angiostrongylus costaricensis were cultured to young adult stages in Waymouth's chemically defined medium MB 752/l, which comprises higher concentrations of the essential components histidine, lysine, methionine, tryptophan, choline chloride and glucose than various other chemically defined media. The present study has shown that choline chloride and tryptophan are required at relatively higher concentrations for worm development than those of the other essential components.


2005 ◽  
Vol 17 (2) ◽  
pp. 193
Author(s):  
G. Horvath ◽  
L. Solti ◽  
G. Seidel

Addition of cholesterol-loaded cyclodextrin (CLC) can increase sperm cryosurvival (Purdy et al. 2000 Cryobiology 48, 36–45). The purpose of this study was to determine if cryosurvival of vitrified oocytes could be improved by incubation with CLC prior to vitrification. Slaughterhouse-derived cumulus oocyte complexes were matured in a chemically defined medium with fatty acid-free BSA and hormones for 21 h followed by partial cumulus removal with 100 U/mL hyaluronidase and gentle pipetting. For an additional hour, oocytes were placed into maturation medium supplemented with 0.5% PVA instead of BSA with or without 2.5 mg/mL CLC. At 22 h after the start of maturation, oocytes were transferred to handling media containing 20% FCS or 0.5% PVA in TCM-199 + HEPES (HTCM-199). Oocytes with approximately 3 layers of cumulus were vitrified in two steps. First, they were exposed to VS1 (10% ethylene glycol (EG), 10% DMSO, 6% PVP, or 20% FCS, in HTCM-199) for 30 s, then exposed to VS2 (20% EG, 20% DMSO, 6% PVP, or 20% FCS, 0.48 M galactose in HTCM-199) for 25 s, loaded into cryoloops in groups of five, and plunged into liquid nitrogen. Rapidly warmed oocytes were moved stepwise through 0.5, 0.25, 0.125, and 0 M galactose in HTCM-199 + 20% FCS, 3 min each. All procedures were conducted at 39°C. Warmed oocytes were placed in maturation medium for an additional hour, fertilized with semen from 3 bulls, 3 replicates each, and cultured according to standard procedures (Zhang et al. 2003 Theriogenology 60, 1657–1663). For each replicate, 30 oocytes were assigned to the following treatments: A: chemically-defined media with PVA for the last hour of maturation, handling and vitrification; B: same as A except CLC treatment, for 1 h before vitrification; C: chemically defined media for maturation, but with 20% FCS for HM, VS1 and VS2. Data were analyzed by ANOVA. CLC treatment resulted in higher cleavage rates and 8- to 16-cell embryo production, but not higher blastocyst (bl) production (Table 1). Non-vitrified oocytes developed better than vitrified ones (means: cleavage, 76%; 8- to 16-cell, 64%; bl D8, 21%; bl D9, 24%). Further studies with vitrification of cholesterol-loaded cyclodextrin-treated oocytes and chemically defined media are warranted. Table 1. Development of vitrified oocytes (LS means ± SE)


Animals ◽  
2021 ◽  
Vol 11 (5) ◽  
pp. 1414
Author(s):  
Josep M. Cambra ◽  
Emilio A. Martinez ◽  
Heriberto Rodriguez-Martinez ◽  
Maria A. Gil ◽  
Cristina Cuello

The development of chemically defined media is a growing trend in in vitro embryo production (IVP). Recently, traditional undefined culture medium with bovine serum albumin (BSA) has been successfully replaced by a chemically defined medium using substances with embryotrophic properties such as platelet factor 4 (PF4). Although the use of this medium sustains IVP, the impact of defined media on the embryonic transcriptome has not been fully elucidated. This study analyzed the transcriptome of porcine IVP blastocysts, cultured in defined (PF4 group) and undefined media (BSA group) by microarrays. In vivo-derived blastocysts (IVV group) were used as a standard of maximum embryo quality. The results showed no differentially expressed genes (DEG) between the PF4 and BSA groups. However, a total of 2780 and 2577 DEGs were detected when comparing the PF4 or the BSA group with the IVV group, respectively. Most of these genes were common in both in vitro groups (2132) and present in some enriched pathways, such as cell cycle, lysosome and/or metabolic pathways. These results show that IVP conditions strongly affect embryo transcriptome and that the defined culture medium with PF4 is a guaranteed replacement for traditional culture with BSA.


Sign in / Sign up

Export Citation Format

Share Document