In vitro Growth of the Nasal Septal Cartilage of the Rat in a Serum-Free Culture Medium

1988 ◽  
Vol 131 (3) ◽  
pp. 231-234 ◽  
Author(s):  
Steinar Kvinnsland
1986 ◽  
Vol 16 (4) ◽  
pp. 387-390 ◽  
Author(s):  
P. Vincendeau ◽  
B. Guillemain ◽  
S. Daulouede ◽  
C. Ripert

Blood ◽  
1974 ◽  
Vol 44 (1) ◽  
pp. 77-85 ◽  
Author(s):  
Allan J. Erslev

Abstract Normal rabbits exposed to 0.4 atmospheric pressure for 3 hr will generate about 40-60 U of erythropoietin during a subsequent 3-hr period. If the kidneys were removed from 3-hr hypoxic animals, washed carefully, and perfused for 3 hr by recirculation with a serum-tissue culture mixture, each kidney generated about 14 U of erythropoietin in vitro. Perfusion of normal kidneys did not result in the production of erythropoietin, and only small amounts were generated if the perfusate contained Puromycin. Three-hour hypoxic kidneys perfused for 3 hr with a serum-free tissue culture medium were found to generate about 8 U of erythropoietin per kidney and similar kidneys perfused with saline about 1 U. These results indicate that erythropoietin is synthesized by kidney tissue and not produced by enzymatic activation of a plasma substrate.


2018 ◽  
Vol 48 (7) ◽  
Author(s):  
Renato Fernandes Galdiano Júnior ◽  
Cibele Mantovani ◽  
Eliana Gertrudes de Macedo Lemos

ABSTRACT: The aim of the present study was to evaluate the effects of carbohydrate supplementation on the propagation of the orchid Cattleya schilleriana. The 120-d-old seedlings were subcultured in fructose-, glucose-, or sucrose-supplemented (0, 15, 30, and 45g L-1) ½ MS culture medium (half-strength macronutrient concentrations), using a completely random design with four repetitions per treatment. After 120d of treatment, root number and length, leaf number and length, and fresh weight were evaluated, and seedling survival was evaluated after 75d of acclimatization in a greenhouse. The in vitro growth data were submitted to regression analysis, whereas the percentage survival data were analyzed using ANOVA and Tukey’s test. Both in vitro growth and ex vitro survival were lowest when the plantlets were grown in the absence of a carbohydrate source and highest (>90% survival) when supplemented with glucose. According to our findings, the addition of either glucose (30g L-1) or sucrose (30g L-1) is recommended for mass propagation of C. schilleriana.


2021 ◽  
Vol 8 ◽  
Author(s):  
Yukina Kuwahara ◽  
Karin Yoshizaki ◽  
Hidetaka Nishida ◽  
Hiroaki Kamishina ◽  
Sadatoshi Maeda ◽  
...  

Mesenchymal stem/stromal cells (MSCs) have been used as cell sources for treating dogs with naturally-occurring diseases. Extracellular vesicles (EVs) derived from MSCs are now recognized as pivotal to modulating the immune response and supporting tissue repair. Manufacture of MSC-EVs for clinical application mandates removal of the xeno-proteins, including fetal bovine serum. The objective of this study was to examine whether canine MSCs survived and secreted EVs in serum-free medium (SFM) conditions and to assess the immunomodulatory effect of EVs in vitro. Canine MSCs were found to survive and secrete EVs under SFM conditions. The surface markers of MSCs in the SFM were similar to MSCs in complete culture medium. Canine MSC-EVs had a diameter of ~300 nm and were positive for EV markers. MSC-derived EVs from the serum-free condition reduced the levels of IL-1β by BV-2 cells in response to LPS stimulation. These results warrant further studies of the use of SFM for producing EVs derived from canine MSCs.


2021 ◽  
Vol 51 ◽  
Author(s):  
Kaliana Gottschalk de Freitas ◽  
José Carlos Sorgato ◽  
Jackeline Schultz Soares ◽  
Luan Marlon Ribeiro

ABSTRACT Some factors may influence the in vitro culture of native orchids, especially the culture medium, sealing system and light condition. This study aimed to evaluate the addition of banana pulp and peel in the formulation of culture media, sealing system and irradiance, in the in vitro growth of Cattleya nobilior Rchb. f. A completely randomized design was used in a factorial scheme with five culture media [Murashige & Skoog supplemented or not with banana pulp or peel (100 or 200g L-1)], two light conditions [3,000 K LED lamps (43 µmol m-2 s-1 or 86 µmol m-2 s-1)] and two sealing systems (natural and conventional ventilation), with six replicates of one vial each. The plant height, diameter of the largest pseudobulb, length of the largest root and of the largest leaf, fresh weight and number of leaves, roots and shoots were evaluated. The use of both the banana pulp and peel as additives in the culture medium formulation promoted the C. nobilior growth. The natural ventilation system increased the plant height, pseudobulb diameter and leaf length, while the conventional system promoted tillering.


2005 ◽  
Vol 17 (2) ◽  
pp. 217 ◽  
Author(s):  
C. Daniaux ◽  
B. Verhaeghe ◽  
I. Donnay

Serum in embryo culture medium may be a potential cause of abnormal accumulation of lipid droplets, which is correlated to a higher sensitivity to cryopreservation. Moreover, serum may introduce pathogens. With the aim of developing a serum-free culture medium, we first (Experiment 1) investigated the effect of adding ITS (5 μg/mL insulin, 5 μg/mL transferrin, 5 ng/mL selenium) as a serum substitute in SOF medium on embryos cultured in large groups (20 embryos per culture drop of 20 μL) and we then (Experiment 2) analyzed the effect of adding BSA. In this second experiment, our serum-free culture media were also tested on embryos cultured in small numbers (5 embryos per drop of 20 μL) in order to mimic ovum pickup (OPU) conditions. Embryos were obtained from slaughterhouse oocytes, matured in vitro for 24 h in a serum-free enriched 199 medium (Donnay et al. 2004 Reprod. Fertil. Dev. 16, 274) containing ITS, and fertilized for 18 h. In experiment 1, embryos were cultured in SOF (Holm et al. 1999 Theriogenology 52, 683–700) supplemented with 0.1 mg/mL polyvinylpyrrolidane (PVP) without (SOF) or with ITS (SOF-ITS), or with 5% FCS (SOF-FCS). Cavitation occurred earlier in presence of serum (Table). Adding ITS to SOF increased blastocyst rates at Day 7 and Day 8 post-insemination (p.i.) and also the hatching rate. In experiment 2, embryos were cultured in SOF-FCS, SOF-ITS, or SOF-ITS supplemented with 4 mg/mL fatty acid free BSA (SOF-ITS-BSA). Within each condition, no differences were observed for blastocyst and hatching rates between embryos cultured in large or in small groups. Adding BSA to SOF-ITS increased blastocyst rate at Day 6 p.i. and also the hatching rate. At Days 7 and 8 p.i., blastocyst rates were higher in SOF-FCS than in SOF-ITS and tended to be higher than in SOF-ITS-BSA, especially for embryos cultured in small groups. Cell numbers of the resulting embryos were unaffected. These results indicate that: (1) ITS as supplement to SOF medium promotes embryo development in vitro. (2) BSA as protein supplement to SOF-ITS medium accelerates blastulation and improves hatching rate. (3) SOF-ITS and SOF-ITS-BSA are two serum-free culture media that can sustain development of embryos, also when cultured in small number, even though SOF-FCS tended to afford better rates of development. Further studies will include evaluation of other quality parameters including resistance to cryopreservation. This work was supported by the Ministery of Agriculture of the Region wallonne de Belgique.


2006 ◽  
Vol 72 (7) ◽  
pp. 5097-5099 ◽  
Author(s):  
Phyllis M. O'Donnell ◽  
Hernan Aviles ◽  
Mark Lyte ◽  
Gerald Sonnenfeld

ABSTRACT Norepinephrine is a stress hormone that enhances bacterial growth. We examined the effects of a small inoculum on the norepinephrine-induced growth of species previously reported to be unaffected by norepinephrine. The results indicated that a reduced inoculum density is essential for observing norepinephrine-induced effects. Additional studies using serum-free media suggested that transferrin plays a role in norepinephrine-induced growth.


2006 ◽  
Vol 18 (2) ◽  
pp. 188
Author(s):  
F. George ◽  
C. Daniaux ◽  
G. Genicot ◽  
F. Focant ◽  
B. Verhaeghe ◽  
...  

In vitro-produced (IVP) bovine blastocysts are known to be more sensitive to cryopreservation than their in vivo counterparts. Removing serum from the culture medium decreases sanitary risk and could improve embryo resistance to cryopreservation by preventing the accumulation of intracellular lipids. Our objectives were to evaluate the lipid content, resistance to cryopreservation, and sex ratio of IVP embryos cultured in a serum-free system. Oocytes from slaughterhouse ovaries were matured in a serum-free enriched medium (Donnay et al. 2004 Reprod. Fertil. Dev. 16, 274) and cultured in 5% O2 in modified SOF supplemented with 5% FCS (FCS) or with insulin-transferrin-selenium (ITS) and 0.1 mg/mL polyvinylpyrrolidone (PVP) (ITS-PVP) or 4 mg/mL BSA (ITS-BSA) (Daniaux et al. 2005 Reprod. Fertil. Dev. 17, 217). Day 5 morulae were stained with the fluorescent dye Nile Red in order to evaluate their lipid content (Genicot et al. 2005 Theriogenology 63, 1181). Day 7 blastocysts (diameter ≥160 µm) were selected, classified according to their size, and frozen in HEPES-SOF containing 1.5 M ethylene glycol, 0.1 M sucrose, and 1.8 mg/mL wheat peptones (George et al. 2002 Reproduction 29, 51). The lipid content was significantly lower in morulae cultured in ITS-BSA compared with the two other media (320 ± 10 arbitrary fluorescence units vs. 383 ± 12 in FCS and 406 ± 10 in ITS-PVP; n = 271; ANOVA2: P < 0.01). After cryopreservation, a higher total hatching rate was found 24 h post-thawing in blastocysts cultured in ITS-BSA and for both serum-free conditions at 48 h (Table 1). In particular, embryos ≤180 µm cultured in FCS were less resistant to cryopreservation than embryos of the same size produced without serum. Expanded blastocysts cultured in ITS-BSA were sexed by PCR (Grisart et al. 1995 Theriogenology 43, 1097) and a higher proportion of male embryos was found (62.7%; n = 51). In conclusion, a complete serum-free system was set up from oocyte maturation to embryo cryopreservation that gave high quality embryos resistant to cryo-preservation. Embryos produced in ITS-BSA presented a lower lipid content, but a shift of the expanded blastocyst sex ratio toward males was observed. Table 1. Hatching rates post-thawing as a function of the blastocyst size and the culture medium


2008 ◽  
Vol 139 (2_suppl) ◽  
pp. P87-P88
Author(s):  
Angela Chang ◽  
Sage August ◽  
Barbara L Schumacher ◽  
Williams Gregory ◽  
Robert L Sah ◽  
...  

Problem Tissue engineering of human nasal septal cartilage represents an alternative technique for creating large quantities of autologous material for use in reconstructive surgery of the head and neck. Septal neocartilage constructs developed in vitro by the alginate method have demonstrated cartilaginous extracellular matrix production, but their biocompatibility and development in vivo remains largely unknown. Methods A murine model was used to examine the behavior of neocartilage constructs in vivo. Chondrocytes collected from donors undergoing septoplasty were expanded in monolayer and suspended in alginate beads for three-dimensional culture in media containing human serum and growth factors. After in vitro incubation for 5 weeks, the neocartilage constructs were implanted subcutaneously in the dorsum of athymic mice for 30 days (n=3). The mice were sacrificed and the constructs were explanted for assessment of cell viability, gross morphology, and histology. Results The mice survived and tolerated the implant well. Infection and extrusion were not observed. Neocartilage constructs maintained their general shape and size, and demonstrated cell viability after implantation. Explanted constructs were firm and opaque, sharing closer semblance to native septal tissue relative to the gelatinous, translucent pre-implant constructs. On hematoxylin and eosin staining, the explanted constructs exhibited distinct morphologies characteristic of native tissue, which were not observed in pre-implant constructs. Conclusion Neocartilage constructs are viable in an in vivo murine model. The morphologic and histologic features of explanted constructs more closely resemble native septal tissue when compared to pre-implant constructs. Significance Septal neocartilage constructs are biocompatible and demonstrate potential for in vivo maturation with eventual clinical application.


2010 ◽  
Vol 106 (5) ◽  
pp. 1249-1252 ◽  
Author(s):  
Igor de Almeida Rodrigues ◽  
Bianca Alcântara da Silva ◽  
André Luis Souza dos Santos ◽  
Alane Beatriz Vermelho ◽  
Celuta Sales Alviano ◽  
...  

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