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Lab on a Chip ◽  
2022 ◽  
Author(s):  
Yoshikazu Kameda ◽  
Surachada Chuaychob ◽  
Miwa Tanaka ◽  
Yang Liu ◽  
Ryu Okada ◽  
...  

Three-dimensional (3D) tissue culture is a powerful tool for understanding physiological events. However, 3D tissues still have limitations in their size, culture period, and maturity, which are caused by the...


2021 ◽  
Vol 22 (23) ◽  
pp. 13080
Author(s):  
Kitaru Suzuki ◽  
Jun Fukasawa ◽  
Maiko Miura ◽  
Poon Nian Lim ◽  
Michiyo Honda ◽  
...  

With the limitation of autografts, the development of alternative treatments for bone diseases to alleviate autograft-related complications is highly demanded. In this study, a tissue-engineered bone was formed by culturing rat bone marrow cells (RBMCs) onto porous apatite-fiber scaffolds (AFSs) with three-dimensional (3D) interconnected pores using a radial-flow bioreactor (RFB). Using the optimized flow rate, the effect of different culturing periods on the development of tissue-engineered bone was investigated. The 3D cell culture using RFB was performed for 0, 1 or 2 weeks in a standard medium followed by 0, 1 or 2 weeks in a differentiation medium. Osteoblast differentiation in the tissue-engineered bone was examined by alkaline phosphatase (ALP) and osteocalcin (OC) assays. Furthermore, the tissue-engineered bone was histologically examined by hematoxylin and eosin and alizarin red S stains. We found that the ALP activity and OC content of calcified cells tended to increase with the culture period, and the differentiation of tissue-engineered bone could be controlled by varying the culture period. In addition, the employment of RFB and AFSs provided a favorable 3D environment for cell growth and differentiation. Overall, these results provide valuable insights into the design of tissue-engineered bone for clinical applications.


2021 ◽  
Vol 75 (1) ◽  
Author(s):  
Baoyi Zhang ◽  
Manyi Li ◽  
Maoguo An ◽  
Chenglong Zhi ◽  
Qingcai Li ◽  
...  

AbstractIron (Fe) is an essential component for marine ecosystems, and it is related to the growth of phytoplankton communities and environmental evolution in coastal area. However, the effect of aquaculture activities on sediment Fe levels is not well studied. Fe levels and grain sizes are determined in two cores (respectively Core C in the culture area and Core A in the control area) in Sishili Bay to reveal the influence of cultivation on sediment Fe levels over an extended period. The sediment Fe levels are distinguished in the upper sections (culture period) but equal in the lower sections (non-culture period) of the two cores. The core C has the same Fe levels as Core A before 1950s (non-culture period). However, the sediment Fe levels of Core C increased during 1950s–1970s (the algae culture period) and decreased after the 1970s (shellfish culture period) compared with Core A, indicating the algae and shellfish culture impose opposite effects on sediment Fe levels. Similarly, sediment grain sizes are observed to be finer during the algae culture period but coarser during the shellfish culture period, and the variation of sediment grain sizes because of culture activities is the important factor affecting sediment Fe levels. The slowing down of ocean current due to algae culture causes finer particles and higher Fe levels in sediment. However, during the shellfish culture period, bio-deposition and re-suspension play major roles in coarsening sediment particles and decreasing sediment Fe levels.


2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Joshua Luchan ◽  
Christian Choi ◽  
Rebecca L. Carrier

AbstractInteractions between epithelial and immune cells with the gut microbiota have wide-ranging effects on many aspects of human health. Therefore, there is value in developing in vitro models capable of performing highly controlled studies of such interactions. However, several critical factors that enable long term homeostasis between bacterial and mammalian cultures have yet to be established. In this study, we explored a model consisting of epithelial and immune cells, as well as four different bacterial species (Bacteroides fragilis KLE1958, Escherichia coli MG1655, Lactobacillus rhamnosus KLE2101, or Ruminococcus gnavus KLE1940), over a 50 hour culture period. Interestingly, both obligate and facultative anaerobes grew to similar extents in aerobic culture environments during the co-culture period, likely due to measured microaerobic oxygen levels near the apical surface of the epithelia. It was demonstrated that bacteria elicited reactive oxygen species (ROS) production, and that the resulting oxidative damage heavily contributed to observed epithelial barrier damage in these static cultures. Introduction of a ROS scavenger significantly mitigated oxidative damage, improving cell monolayer integrity and reducing lipid peroxidation, although not to control (bacteria-free culture) levels. These results indicate that monitoring and mitigating ROS accumulation and oxidative damage can enable longer term bacteria-intestinal epithelial cultures, while also highlighting the significance of additional factors that impact homeostasis in mammalian cell-bacteria systems.


Animals ◽  
2021 ◽  
Vol 11 (12) ◽  
pp. 3347
Author(s):  
Konstantina Stamperna ◽  
Themistoklis Giannoulis ◽  
Eleni Dovolou ◽  
Maria Kalemkeridou ◽  
Ioannis Nanas ◽  
...  

The aims of the present study were to examine the effects of HSP70 addition in the in vitro culture medium of day 3 embryos on their developmental competence and quality. Bovine oocytes (n = 1442) were in vitro matured, inseminated and cultured for the first two days according to standardized methods. The presumptive zygotes were randomly allocated in three experimental groups: Control, C (embryos cultured at 39 °C throughout the culture period), group C41 (temperature was raised to 41 °C from the 48th to 72nd h post insemination (p.i.) and then it returned at 39 °C for the remaining culture period), and group H41 (the temperature modification was the same as in C41 and during heat exposure, HSP70 was added in the culture medium). Cleavage and embryo yield were assessed 48 h p.i. and on days 7, 8, 9, respectively and gene expression in day 7 blastocysts was assessed by RT-PCR. Blastocyst yield was the highest in group C39; and higher in group H41 compared to group C41. From the gene expression analyses, altered expression of 11 genes was detected among groups. The analysis of the orchestrated patterns of gene expression differed between groups. The results of this study confirm the devastating effects of heat stress on embryo development and provide evidence that HSP70 addition at the critical stages can partly counterbalance, without neutralizing, the negative effects of the heat insult on embryos, acting mainly through mechanisms related to energy deployment.


Plant Methods ◽  
2021 ◽  
Vol 17 (1) ◽  
Author(s):  
Bo-Kook Jang ◽  
Ju-Sung Cho ◽  
Shin-Ho Kang ◽  
Cheol Hee Lee

Abstract Background Liquid suspension culture efficiently proliferates plant cells and can be applied to ferns because it rapidly increases the fresh weight of gametophytes. This study assessed gametophyte proliferation and sporophyte production of Pteridium aquilinum var. latiusculum using a suspension culture method. Results The growth curve linear phase of gametophyte cells was confirmed between 9 and 18 days of culture, and the subculture cycle was determined to be 2 weeks. A double-strength MS medium (fresh weight, 18.0 g) containing 2% sucrose and NH4+:NO3− (120 mM, 40:80) was found to be the optimal liquid medium. Gametophytes obtained after suspension culture for 18 days did not normally form sporophytes in an ex vitro soil environment. However, this issue was resolved after changing the culture type or extending the culture period to 6 weeks. A short suspension culture period increased the fresh weight of fragmented and homogenized gametophytes but yielded numerous relatively immature gametophytes (globular forms of branching gametophytes, BG). Furthermore, differences in gametophyte morphogenesis and development were indicated by changes in endogenous phytohormone content. BG with immature development exhibited high accumulation of zeatin, jasmonic acid, and salicylic acid, and relatively low levels of abscisic acid and indole-3-acetic acid. The immature development of gametophytes directly affected sporophyte formation. Conclusions This study maximized the advantages of liquid suspension culture using eastern bracken gametophytes and provides data to resolve any associated issues, thus facilitating efficient bracken production.


2021 ◽  
Vol 6 (13) ◽  
pp. 91-100
Author(s):  
Esra Balcioglu ◽  
Munevver Baran ◽  
Mehtap Nisari ◽  
Ozge Goktepe ◽  
Pinar Bilgici ◽  
...  

Background: Ionizing radiation poses a threat to the early embryo possibly leading to prenatal death, growth retardation, organ malformation, or mental retardation. It is important, assessment of any adverse effects of radiation upon the embryo. This study aimed to evaluate the outcomes of embryos irradiated with 1Gy doses in vitro and investigate hematopoiesis in the yolk sac of the irradiated embryos. Materials and methods: In the study, the experimental group of rats was be exposed to total body ionizing radiation on days 8.5th of gestation. All embryos in the control and radiation group cultured from gestation day 9.5 to 11.5 were alive at the end of the culture period. After 48 hours culture period, the embryos from each group were harvested and analyzed morphologically. Histological evaluation of the vWF+ cell number was performed in vivo. Results: The results showed that the embryonic growth and development during organogenesis decreased in the radiation exposed embryos when compared to control embryos. Additionally, the immunofluorescent examination showed that the vWF+ cell number reduced in the yolk sac of embryos exposed to ionizing radiation. Conclusion: Consequently, these findings support the conclusion that 1 Gy ionizing irradiation may increase prenatal death, intrauterine growth restriction on embryonic development when ionizing irradiation decreases the vWF+ cell number in the yolk sac compared to control embryos. This research related to radiation was the first study using the in vitro embryo culture technique; thus, future studies that will be performed by using different doses of radiation will contribute to the literature.


Author(s):  
Deepa Pathipati ◽  
Srinivasa Prasad Chigurupati ◽  
A.V.N. Siva Kumar ◽  
B. Punya Kumari ◽  
R.V. Suresh Kumar

Background: In in vitro culture systems IGF-I was known to promote follicular maturation, granulosa cell proliferation and increase overall cellular function. The expression patterns of developmentally important markers for IGF-I are expressed differentially during the transition of preantral follicles to the ovulatory stage. Therefore, we aimed to investigate the influence of time specific addition of Insulin like growth factor-I (IGF-I) on in vitro development of sheep preantral follicles (PFs’). Methods: Mechanically isolated sheep preantral follicles were cultured for six days either in TCM 199B or standard medium supplemented with IGF-I at different points during the culture period and were subsequently subjected to in vitro maturation (IVM) for additional 24hrs to evaluate different follicular development parameters. Conclusion: Based on the results, it is concluded that: 1) IGF-I supplementation was needed during early rather than later stages of culture period, therefore, role of IGF-I in time/stage specific manner in cultured preantral follicles is confirmed. 2) Supplementation of TCM 199B with IGF-I simultaneously for the first two days followed by TCM 199B alone without any growth factor (s) in later days (Treatment 1) supported better development of PFs’ in vitro.


Author(s):  
Shahla Babaki ◽  
Saeed Zavareh ◽  
Parisa Farrokh ◽  
Meysam Nasiri

Background: Wnt signaling pathway plays critical role in ovarian follicle development. Therefore, the aim of this study was to evaluate the effects of vitrification on the expression of Wnt pathway related genes in preantral follicles (PFs). Methods: Isolated PFs (n=982) of 14-16 day old female mice (n=45: 15 for each group) were divided into fresh (n=265), toxicity (n=272), and vitrified (n=265). The mRNA levels of Wnt2, Wnt4, Lrp5 and Fzd3 were evaluated by real-time PCR on the 2nd and 6th days of culture period. One-way ANOVA was conducted to analyze the data. Post hoc Tukey's HSD was used for multiple comparisons and p-value less than 0.05 was considered statistically significant. Results: The developmental parameters of fresh PFs were significantly higher than those of vitrified (p<0.001). There were no differences between fresh and vitrified PFs on the 2nd day of culture (p<0.001). Wnt4 expression levels decreased significantly in vitrified groups compared with fresh ones (p<0.001). Fzd3 and Lrp expression levels increased significantly in vitrified groups compared with those in the fresh group on the 2nd day (p<0.001). On the 6th day of culture period, the expression levels of Wnt2 and Fzd3 increased significantly in vitrified group compared to those of fresh group (p<0.001). Moreover, the expression levels of Wnt4 and Lrp increased significantly in toxicity groups compared to those of the control group (p<0.001). Conclusion: Vitrification increase the expression levels of Wnt2, Lrp and Fzd3 genes of PFs during in vitro culture


Author(s):  
Alish Debbarma ◽  
Stephen Sampath Kumar ◽  
J. V. Senthil Kumar ◽  
Anupam Tripura

The study was conducted to evaluate the effect of stocking density influencing the growth of Hybrid Tilapia (Red strain) in cages for culture period of 120 days. The experiment is conducted in Thanjavur Center for sustainable Aquaculture, Sorrokottai, Tamil Nadu, India. Hybrid Tilapia fingerling ABW of (3.3+0.01 g) were stocked at 20/m3 (T1), 30/m3 (T2) and 40/m3 (T3) and fed 3 times daily (9:00, 12:00 and 17:00 H) with a commercial food 40% protein gradually reduce to 35 to 30% protein and stock was sampled fortnight. Among three stocking densities, 20 fish/m3 was found to be the best for the growth in cages, while the FCR and net biomass production were found higher in high stocking densities fishes (30, 40 fish/m3). The best FCR was 1.08 +0.06 from T1 (20 fish/m3) with 92.5+0.04% of FCE. The highest net biomass production was 19 kg (3.16 kg/m3) from T2 (30 fish/m3). Present study reveal that stocking density has negative effect on bio-growth parameters but biomass production has positive effect with stocking density.


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