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We evaluated the inhibitory effects of the probiotic Lactobacillus species on different phases of Candida albicans biofilm development. Quantification of biofilm growth and ultrastructural analyses were performed on C. albicans biofilms treated with Lactobacillus rhamnosus , Lactobacillus casei , and Lactobacillus acidophilus planktonic cell suspensions as well as their supernatants. Planktonic lactobacilli induced a significant reduction ( p  < 0.05) in the number of biofilm cells (25.5–61.8 %) depending on the probiotic strain and the biofilm phase. L. rhamnosus supernatants had no significant effect on the mature biofilm ( p  > 0.05), but significantly reduced the early stages of Candida biofilm formation ( p  < 0.01). Microscopic analyses revealed that L. rhamnosus suspensions reduced Candida hyphal differentiation, leading to a predominance of budding growth. All lactobacilli negatively impacted C. albicans yeast-to-hyphae differentiation and biofilm formation. The inhibitory effects of the probiotic Lactobacillus on C. albicans entailed both cell-cell interactions and secretion of exometabolites that may impact on pathogenic attributes associated with C. albicans colonization on host surfaces and yeast filamentation. This study clarifies, for the first time, the mechanics of how Lactobacillus species may antagonize C. albicans host colonization. Our data elucidate the inhibitory mechanisms that define the probiotic candicidal activity of lactobacilli, thus supporting their utility as an adjunctive therapeutic mode against mucosal candidal infections.

Cell dielectric property measurement holds significant potential for application in cell detection and diagnosis due to its label-free and noninvasive nature. In this study, we developed a biosensor designed to measure the permittivity of liquid samples, particularly cell suspensions at the nanoliter scale, utilizing microwave and millimeter wave coplanar waveguides in conjunction with a microchannel. This biosensor facilitates the measurement of scattering parameters within a frequency domain ranging from 1 GHz to 110 GHz. The obtained scattering parameters are then converted into dielectric constants using specific algorithms. A cell capture structure within the microchannel ensures that cell suspensions remain stable within the measurement zone. The feasibility of this biosensor was confirmed by comparison with a commercial Keysight probe. We measured the dielectric constants of three different cell suspensions (HepG2, A549, MCF-7) using our biosensor. We also counted the number of cells captured in multiple measurements for each cell type and compared the corresponding changes in permittivity. The results indicated that the real part of the permittivity of HepG2 cells is 0.2–0.8 lower than that of the other two cell types. The difference between A549 and MCF-7 was relatively minor, only 0.2–0.4. The fluctuations in the dielectric spectrum caused by changes in cell numbers during measurements were smaller than the differences observed between different cell types. Thus, the sensor is suitable for measuring cell suspensions and can be utilized for label-free, noninvasive studies in identifying biological cell suspensions.

Plant secondary biosynthetic pathways are exceedingly inducible by elicitors and facilitate enhanced metabolite production using plant cell tissue and organ cultures. Elicitors can regulate large number of control points and trigger the expression of key genes with increased cellular activities at biochemical and molecular level involving signal compounds. A large number of chemical elicitors viz: jasmonic acid (JA), methyl jasmonate (MeJA), salicylic acid (SA), acetyl salicylic acid (ASA), ethylene (ET) and ethrel (Ethe), heavy metals (HM), many types of chemical compounds (natural or synthetic) and their combinations are used for elicitation studies. Cell suspensions and hairy roots are commonly used culture systems followed by adventitious roots and multiple shoots for elicitation experiments. Amongst the elicitors and concentrations used 100 µM MeJA was found optimum for secondary metabolite enhancement in majority of experiments compared to SA and JA. Elicitor treatments promoted yield enhancements starting from 1.0 to maximum of 2230-fold across plant species studied. Elicitors singly with media additives, combination of elicitors and elicitors other than signal compounds along with hormones were found beneficial for enhanced secondary metabolite production. Further, a combination of target gene over expression and elicitor treatment also supported higher secondary product yield. The present communication presents information exclusively about the use of chemical elicitors for secondary metabolites production in vitro covering approximately more than a decade of research at one place in one review. Further, this extensive appraisal will be useful for the understanding and manipulation of secondary metabolites for enhanced production in vitro.

Although zebrafish are commonly used to study intestinal mucosal immunity, no dedicated procedure for isolating immune cells from zebrafish intestines is currently available. A speedy and simple operating approach for preparing cell suspension from mucosa has been devised to better understanding of intestinal cellular immunity in zebrafish. The mucosal villi were separated away from the muscle layer by repeated blows. The complete deprivation of mucosa was done and evidenced by HE and results. Higher expression of both innate ( , , and ) and adaptive immune genes ( , , , and ) was revealed compared to cells obtained by typical mesh rubbing. The cytometric results also revealed that the tested operation group had a higher concentration and viability. Further, fluorescent-labelled immune cells from 3mo , , , and , were isolated and evaluated for the proportion, and immune cells' type could be inferred from the expression of marker genes. The transcriptomic data demonstrated that the intestinal immune cell suspension made using the new technique was enriched in immune-related genes and pathways, including , and , as well as pattern recognition receptor signaling and cytokine-cytokine receptor interaction. In addition, the low expression of DEG for the adherent and close junctions indicated less muscular contamination. Also, lower expression of gel-forming mucus-associated genes in the mucosal cell suspension was consistent with the current less viscous cell suspension. To apply and validate the developed manipulation, enteritis was induced by soybean meal diet, and immune cell suspensions were analyzed by flow cytometry and qPCR. The finding that in enteritis samples, there was inflammatory increase of neutrophils and macrophages, was in line with upregulated cytokines ( and ) and cell markers ( and ). As a result, the current work created a realistic technique for studying intestinal immune cells in zebrafish. The immune cells acquired may aid in further research and knowledge of intestinal illness at the cellular level.

Background Successful usage of autologous skin cell suspension (ASCS) has been demonstrated in some clinical trials. However, its efficacy and safety have not been verified. This latest systematic review and meta‐analysis aim to examine the effects of autologous epidermal cell suspensions in re‐epithelialization of skin lesions. Methods Relevant articles were retrieved from PubMed, Embase, Cochrane Database, Web of Science, International Clinical Trials Registry Platform, China National Knowledge Infrastructureris, VIP Database for Chinese Technical Periodicals and Wanfang database. The primary output measure was the healing time, and the secondary outputs were effective rate, size of donor site for treatment, size of study treatment area, operation time, pain scores, repigmentation, complications, scar scale scores and satisfaction scores. Data were pooled and expressed as relative risk (RR), mean difference (MD) and standardized mean difference (SMD) with a 95% confidence interval (CI). Results Thirty‐one studies were included in this systematic review and meta‐analysis, with 914 patients who received autologous epidermal cell suspensions (treatment group) and 883 patients who received standard care or placebo (control group). The pooled data from all included studies demonstrated that the treatment group has significantly reduced healing time (SMD = −0.86; 95% CI: −1.59–0.14; p = 0.02, I2 = 95%), size of donar site for treatment (MD = −115.41; 95% CI: −128.74–102.09; p<0.001, I2 = 89%), operation time (MD = 25.35; 95% CI: 23.42–27.29; p<0.001, I2 = 100%), pain scores (SMD = −1.88; 95% CI: −2.86–0.90; p = 0.0002, I2 = 89%) and complications (RR = 0.59; 95% CI: 0.36–0.96; p = 0.03, I2 = 66%), as well as significantly increased effective rate (RR = 1.20; 95% CI: 1.01–1.42; p = 0.04, I2 = 77%). There were no significant differences in the size of study treatment area, repigmentation, scar scale scores and satisfaction scores between the two groups. Conclusion Our meta‐analysis showed that autologous epidermal cell suspensions is beneficial for re‐epithelialization of skin lesions as they significantly reduce the healing time, size of donar site for treatment, operation time, pain scores and complications, as well as increased effective rate. However, this intervention has minimal impact on size of treatment area, repigmentation, scar scale scores and satisfaction scores.

The large-scale production of plant-derived secondary metabolites (PDSM) in bioreactors to meet the increasing demand for bioactive compounds for the treatment and prevention of degenerative diseases is nowadays considered an engineering challenge due to the large number of operational factors that need to be considered during their design and scale-up. The plant cell suspension culture (CSC) has presented numerous benefits over other technologies, such as the conventional whole-plant extraction, not only for avoiding the overexploitation of plant species, but also for achieving better yields and having excellent scaling-up attributes. The selection of the bioreactor configuration depends on intrinsic cell culture properties and engineering considerations related to the effect of operating conditions on thermodynamics, kinetics, and transport phenomena, which together are essential for accomplishing the large-scale production of PDSM. To this end, this review, firstly, provides a comprehensive appraisement of PDSM, essentially those with demonstrated importance and utilization in pharmaceutical industries. Then, special attention is given to PDSM obtained out of CSC. Finally, engineering aspects related to the bioreactor configuration for CSC stating the effect of the operating conditions on kinetics and transport phenomena and, hence, on the cell viability and production of PDSM are presented accordingly. The engineering analysis of the reviewed bioreactor configurations for CSC will pave the way for future research focused on their scaling up, to produce high value-added PDSM.

The aim of this study is to determine the optimal hormone concentration and suspension culture conditions for callus induction in Cassia mimosoides herb. The leaves of aseptic seedlings were used as explants to determine the effects of various concentrations of 6-BA (6-benzylamino purine) and NAA (1-naphthaleneacetic acid) on the callus induction rate through single-factor experiments. The study examined the effect of initial suspension cell inoculation and inoculum on cell biomass. Using a hormone concentration combination of 3.0 mg L-1 BA and 1.0 mg L-1 NAA, the callus induction rate of the tender leaves of the aseptic seedlings reached 100% with good callus status. In single-factor suspension cell culture experiments, an initial inoculum of 2 g and an inoculum volume of 2 mL were most conducive to the increase in cell biomass. The content of emodin in the suspension cells and the original herb of Cassia mimosoides herb was measured by high-performance liquid chromatography (HPLC). The results indicated that at the same mass, the content of emodin in the suspension cells was 12.22-fold higher compared with that in the original herb. The results indicate that the suspension cells cultured by this method can increase the content of emodin in Cassia mimosoides herb, thus providing a more convenient and efficient method to extract emodin. The small cell clusters and single cells obtained in suspension culture can be directly used for protoplast isolation and also serve as direct receptors for genetic transformation. This provides a foundation for large-scale cell culture and the genetic transformation of Cassia mimosoides herb suspension culture.

Taxodium hybrid ‘zhongshanshan’ is an improved variety with significant applications in landscaping, ecological construction and beach afforestation in China, which has a high market demand. Although somatic embryogenesis system of Taxodium hybrid ‘zhongshanshan’ has been successfully established, it currently lacks automation and large-scale production capabilities. Therefore, we urgently need a method that can increase the output of somatic embryos (SEs) to meet the demands of large-scale production. In this study, we utilized embryogenic tissue (ET) of Taxodium hybrid ‘zhongshanshan’ as research material to evaluate the effects of initial ET culture time, inoculum density, suspension culture period and orbiting speed on the suspension culture. We observed that the proliferation of ET exhibited an S-shaped curve, with the exponential phase occurring approximately on the 9th day. The cell proliferation and somatic embryogenesis efficiency were influenced by the inoculum density and suspension culture period of ET, reaching their maximum when the inoculum density was 60 mg mL −1 and suspension cultured for 1 week. Additionally, an orbiting speed of 110 rpm proved to be optimal for our suspension culture, yielding the highest fresh weight (FW), dry weight (DW), higher settled cell volume (SCV), and a well-developed embryonic suspensor mass (ESM) structure. The investigation of this suspension culture system provides valuable insights and foundations for further automation and scale culture of somatic embryogenesis in Taxodium hybrid ‘zhongshanshan’. Key message The present study successfully established an efficient embryogenic cell suspension culture system of Taxodium hybrid ‘zhongshanshan’ for the first time.

Rice cell suspension culture (RCSC) is one of the most widely studied plant cell culture systems next to tobacco and carrot. Simple cell culture techniques, scalability and high genetic transformation potential make RCSC an ideal platform to produce high-value recombinant proteins and plant specialized metabolites (PSM). Our understanding of the rice genome and its genetic regulation makes RCSC amenable to efficient genetic engineering with precision genome editing tools such as CRISPR/Cas. Further, the metabolic pool of RCSC can be harnessed and bioengineered to produce recombinant proteins and PSM. This review highlights the studies performed on transgenic RCSC and potential of this platform to synthesize PSM. Recent advancements in RCSC-mediated production and yield enhancement of bioactives using precision molecular biology tools such as CRISPR/Cas, media optimization and challenges associated with the establishment of RCSC are also summarized. This effort is to put the spotlight back on RCSC, which can become an attractive alternative to existing transgenic plant cell suspension culture systems.Key MessageNew developments in transgenic rice cell suspension culture and subsequent bioprocess optimization have potential to advance the field of plant-based biopharmaceutical production. Further use of precise genetic engineering tools can leverage the prospects of rice cell suspension culture for molecular pharming.

Elicitors trigger complex signaling cascades within plants, enhancing secondary metabolite production. The present study investigated the impact of various elicitors on the augmentation of picrosides (P-I, P-II, P-III) and their precursors (vanillic acid, caffeic acid, cinnamic acid, catalpol, and aucubin) metabolites in Picrorhiza kurroa cell suspension cultures. Four elicitors; methyl jasmonate (Me-JA; 50–150 µM), salicylic acid (SA; 50–150 mg/L), chitosan (CHT; 50–150 mg/L), and yeast extract (YE; 50–150 mg/L) were applied to leaf (LFSC) and rhizome (RHSC) cell suspensions on the 21st day of culture and incubated up to 192 h. In LFSC, the highest contents of P-I (7.90 mg/g DW) and P-III (0.69 mg/g DW) were at Me-JA 150 µM, while P-II (3.99 mg/g DW) was at Me-JA 50 µM. YE 150 mg/L increased vanillic acid (0.29 mg/g DW), and Me-JA 100 µM increased caffeic acid (0.06 mg/g DW) and cinnamic acid (0.19 mg/g DW). CHT 100 mg/L maximized catalpol (5.91 mg/g DW), and aucubin (0.29 mg/g DW) peaked with Me-JA 50 µM, YE 150 mg/L, and CHT 150 mg/L. In RHSC, Me-JA 150 µM yielded the highest levels of P-I (7.74 mg/g DW), P-II (4.59 mg/g DW), cinnamic acid (0.19 mg/g DW), and aucubin (0.29 mg/g DW). The study indicated that Me-JA is particularly effective in enhancing picrosides accumulation in P. kurroa cell cultures. In conclusion, elicitors significantly boost the production of valuable secondary metabolites, particularly in plant cell culture of P. kurroa .