Albumin has a long historical involvement in design of media for the successful culture of mammalian cells, in both the research and commercial fields. The potential application of albumins, bovine or human serum albumin, for cell culture is a by-product of the physico-chemical, biochemical and cell-specific properties of the molecule. In this review an analysis of these features of albumin leads to a consideration of the extracellular and intracellular actions of the molecule, and importantly the role of its interactions with numerous ligands or bioactive factors that influence the growth of cells in culture: these include hormones, growth factors, lipids, amino acids, metal ions, reactive oxygen and nitrogen species to name a few. The interaction of albumin with the cell in relation to these co-factors has a potential impact on metabolic and biosynthetic activity, cell proliferation and survival. Application of this knowledge to improve the performance in manufacturing biotechnology and in the emerging uses of cell culture for tissue engineering and stem cell derived therapies is an important prospect.

/pdf/albumin-and-mammalian-cell-culture-implications-for-kyuqrl6j0d.pdf

Albumin and mammalian cell culture: implications for biotechnology applications

Ultrathin chitosan-poly(ethylene glycol) hydrogel films for corneal tissue engineering.

Heterochronic blood sharing rejuvenates old tissues, and most of the studies on how this works focus on young plasma, its fractions, and a few youthful systemic candidates. However, it was not formally established that young blood is necessary for this multi-tissue rejuvenation. Here, using our recently developed small animal blood exchange process, we replaced half of the plasma in mice with saline containing 5% albumin (terming it a "neutral" age blood exchange, NBE) thus diluting the plasma factors and replenishing the albumin that would be diminished if only saline was used. Our data demonstrate that a single NBE suffices to meet or exceed the rejuvenative effects of enhancing muscle repair, reducing liver adiposity and fibrosis, and increasing hippocampal neurogenesis in old mice, all the key outcomes seen after blood heterochronicity. Comparative proteomic analysis on serum from NBE, and from a similar human clinical procedure of therapeutic plasma exchange (TPE), revealed a molecular re-setting of the systemic signaling milieu, interestingly, elevating the levels of some proteins, which broadly coordinate tissue maintenance and repair and promote immune responses. Moreover, a single TPE yielded functional blood rejuvenation, abrogating the typical old serum inhibition of progenitor cell proliferation. Ectopically added albumin does not seem to be the sole determinant of such rejuvenation, and levels of albumin do not decrease with age nor are increased by NBE/TPE. A model of action (supported by a large body of published data) is that significant dilution of autoregulatory proteins that crosstalk to multiple signaling pathways (with their own feedback loops) would, through changes in gene expression, have long-lasting molecular and functional effects that are consistent with our observations. This work improves our understanding of the systemic paradigms of multi-tissue rejuvenation and suggest a novel and immediate use of the FDA approved TPE for improving the health and resilience of older people.

/pdf/rejuvenation-of-three-germ-layers-tissues-by-exchanging-old-3kpn804pz8.pdf

Rejuvenation of three germ layers tissues by exchanging old blood plasma with saline-albumin

Purpose To examine the molecular profile of fetal human retinal progenitor cells (hRPCs) expanded in vitro and those grown in a co-culture system with mouse retina through the analysis of protein and gene expression and neurotransmitter-stimulated calcium dynamics. Methods hRPCS were isolated from human retina of 14 to 18 weeks gestational age (GA) and expanded in vitro. Immunoblot, microarray, and immunocytochemistry (ICC) assays were performed on undifferentiated hRPCs and those co-cultured with mouse retinas for 2 weeks. Cell function was assessed by using calcium imaging. Results The ICC results showed a gradual decrease in the percentages of KI67-, SOX2-, and vimentin-positive cells from passages (P) 1 to P6, whereas a sustained expression of nestin and PAX6 was observed through P6. Microarray analysis of P1 hRPCs showed the expression of early retinal developmental genes: VIM (vimentin), KI67, NES (nestin), PAX6, SOX2, HES5, GNL3, OTX2, DACH1, SIX6, and CHX10 (VSX2). At P6, hRPCs continued to express VIM, KI67, NES, PAX6, SOX2, GNL3, and SIX6. On co-culture, there was a significant increase in the expression of MKI67, PAX6, SOX2, GNL3, SIX3, and RHO (rhodopsin). Calcium imaging showed a functional response to excitatory neurotransmitters. Conclusions Fetal-derived hRPCs show molecular characteristics indicative of a retinal progenitor state up to P6 (latest passage studied). They show a progressive decrease in the expression of immature markers as they reach P6. These cells are functional, respond to excitatory neurotransmitters, and exhibit changes in expression patterns in response to co-culture with mouse retina.

Molecular characterization of human retinal progenitor cells.

Intact vitreous humor as a potential extracellular matrix hydrogel for cartilage tissue engineering applications.

Intravitreal injection is an important delivery route for studies involving the transplantation of various types of precursor cells to the retina; however, the effect on these cells of exposure to the vitreous microenvironment has not been specifically investigated. Here vitreous humor was evaluated for the potential to influence the proliferation of rat retinal precursor cells in vitro. Cells were isolated at embryonic day 19 and plated in standard proliferation medium in the presence or absence of fluid expressed from porcine vitreous humor. Cellular proliferation at different concentrations of vitreous fluid supplementation was quantified by using a (3)H-thymidine incorporation assay. Active components of vitreous fluid were partially characterized by gel filtration chromatography (GFC) and UV spectral analysis. The effect of each vitreous fraction on proliferation was determined as well. Results showed that addition of 20% vitreous fluid to primary rat retinal cultures significantly increased (3)H-thymidine incorporation compared with growth medium without vitreous supplementation. A vitreous fraction showing growth-promoting activity was localized to a molecular mass range <1000 Da, consistent with ascorbic acid. Ascorbic acid was confirmed in vitreous fluid by UV spectral analysis. Growth-augmenting activity was present in higher molecular mass vitreous fractions, consistent with protein components. Albumin, the major protein in vitreous fluid, was found to augment proliferation. Because vitreous-associated augmentation of retinal precursor proliferation remains an epidermal growth factor-dependent phenomenon, the proliferative status of transplanted cells in the vitreous cavity is likely determined by a combination of factors.

Vitreous humor and albumin augment the proliferation of cultured retinal precursor cells.

Objective To characterize embryonic rat retinal progenitor cells (RPCs) by flow cytometry (FACS), immunofluorescence and (3)H-Thymidine assay in vitro. Methods RPCs were prepared from the retina of embryonic day 19 Sprague Dawley rats and were cultured in DMEM: F12 medium with N2-supplement, epidermal growth factor (EGF), basic fibroblast growth factor (bFGF) and leukemia inhibitory factor (LIF) for 0 to 4 days. Cell proliferation and cluster formation were quantified by (3)H-Thymidine assay and morphometric analysis, respectively. Antibodies used to characterize the RPCs included markers for neural progenitors (Nestin and Sox-2), astrocytes (glial fibrillary acidic protein [GFAP]), horizontal cells (Calretinin), ganglion cells (CD90), rods (Rhodopsin), biopolar cells (PKC) and amacrine cells (Syntaxin) by the use of FACS. Results In primary cells, 22.23%, 16.04% and 19.66% of cells were found to express Nestin, Sox-2 and Calretinin, respectively. GFAP, CD90, Rhodopsin, PKC, Syntaxin were not detected or trail. Meanwhile, more than 90% cells were Nestin positive by immunofluorescence. After 6 days culture in vitro, 43.36% cells express Nestin by FACS. Nestin and GFAP were expressed by use of immunofluorescence. There was significant proliferation of RPCs within the first 4 days as evaluated by (3)H-Thymidine assay. Conclusions The results demonstrate that combine FACS, immunofluorescence and (3)H-Thymidine assay together could characterize RPCs. RPCs proliferate well at the early period of cell culture.

[Isolation, culture and identification of rat retinal progenitor cells in vitro].

Background POD places a heavy burden on the healthcare system as the number of elderly people undergoing surgery is increasing annually because of the aging population. As a large country with a severely aging population, China's elderly population has reached 267 million. There has been no summary analysis of the pooled incidence of POD in the elderly Chinese population. Methods Systematic search databases included PubMed, Web of Science, EMBASE, Cochrane Library Databases, China Knowledge Resource Integrated Database (CNKI), Chinese Biomedical Database (CBM), WanFang Database, and Chinese Science and Technology Periodicals (VIP). The retrieval time ranged from the database's establishment to February 8, 2023. The pooled incidence of delirium after non-cardiac surgery was calculated using a random effects model. Meta-regression, subgroup, and sensitivity analyses were used to explore the source of heterogeneity. Results A total of 52 studies met the inclusion criteria, involving 18,410 participants. The pooled incidence of delirium after non-cardiac surgery in the elderly Chinese population was 18.6% (95% CI: 16.4–20.8%). The meta-regression results revealed anesthesia method and year of publication as a source of heterogeneity. In the subgroup analysis, the gender subgroup revealed a POD incidence of 19.6% (95% CI: 16.9–22.3%) in males and 18.3% (95% CI: 15.7–20.9%) in females. The year of publication subgroup analysis revealed a POD incidence of 20.3% (95% CI: 17.4–23.3%) after 2018 and 14.6 (95% CI: 11.6–17.6%) in 2018 and before. In the subgroup of surgical types, the incidence of hip fracture surgery POD was 20.7% (95% CI: 17.6–24.3%), the incidence of non-cardiac surgery POD was 18.4% (95% CI: 11.8–25.1%), the incidence of orthopedic surgery POD was 16.6% (95% CI: 11.8–21.5%), the incidence of abdominal neoplasms surgery POD was 14.3% (95% CI: 7.6–21.1%); the incidence of abdominal surgery POD was 13.9% (95% CI: 6.4–21.4%). The anesthesia methods subgroup revealed a POD incidence of 21.5% (95% CI: 17.9–25.1%) for general anesthesia, 15.0% (95% CI: 10.6–19.3%) for intraspinal anesthesia, and 8.3% (95% CI: 10.6–19.3%) for regional anesthesia. The measurement tool subgroup revealed a POD incidence of 19.3% (95% CI: 16.7–21.9%) with CAM and 16.8% (95% CI: 12.6–21.0%) with DSM. The sample size subgroup revealed a POD incidence of 19.4% (95% CI: 16.8–22.1%) for patients ≤ 500 and 15.3% (95% CI: 11.0–19.7%) for patients > 500. The sensitivity analysis suggested that the pooled incidence of postoperative delirium in this study was stable. Conclusion Our systematic review of the incidence of delirium after non-cardiac surgery in elderly Chinese patients revealed a high incidence of postoperative delirium. Except for cardiac surgery, the incidence of postoperative delirium was higher for hip fracture surgery than for other types of surgery. However, this finding must be further explored in future large-sample studies. Systematic review registration https://www.crd.york.ac.uk/prospero/, identifier: PROSPERO CRD42023397883.

/pdf/incidence-of-delirium-after-non-cardiac-surgery-in-the-2dx18c8s.pdf

Incidence of delirium after non-cardiac surgery in the Chinese elderly population: a systematic review and meta-analysis

Aqueous humor has been shown to influence the proliferation of various ocular cell types, but the effect on immature retinal cells is not known. Here, the effect of pig aqueous humor on the proliferation of rat retinal precursor cells (RPCs) was investigated. RPCs were prepared from embryonic day 19 Sprague-Dawley rats and cultured in the presence or absence of aqueous humor from healthy pigs along with a medium consisting of Dulbecco's modified Eagle's medium:Ham's F-12 medium, N2 supplement, and epidermal growth factor. Proliferation was quantified by [(3)H]thymidine incorporation under different treatment conditions, and any associated morphological changes were noted. Potential active components of porcine aqueous humor were partially characterized by gel filtration chromatography, and the effect on RPC proliferation was determined. Results showed that adding 20% aqueous humor increased [(3)H]thymidine incorporation by as much as 317%, as compared with controls. Aqueous supplementation also increased both the number and size of RPC spherical aggregates ("spheres") over the first 4 days, consistent with increased proliferative activity. Using gel filtration and the in vitro proliferation assay, the growth-promoting activity of aqueous humor was localized to two different molecular mass ranges, namely, around 30 kDa and less than 1 kDa. Ascorbic acid was present in the lower molecular mass fraction, and use of this molecule reproduced some, but not all, of the proliferative activity present in aqueous humor. These results highlight the potential role of soluble factors present in the cellular microenvironment with respect to modulation of endogenous progenitor cell activity.

https://stemcellsjournals.onlinelibrary.wiley.com/doi/pdf/10.1634/stemcells.2006-0103

Aqueous humor enhances the proliferation of rat retinal precursor cells in culture, and this effect is partially reproduced by ascorbic acid.

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Vitreous humor and albumin augment the proliferation of cultured retinal precursor cells.

[Isolation, culture and identification of rat retinal progenitor cells in vitro].

Incidence of delirium after non-cardiac surgery in the Chinese elderly population: a systematic review and meta-analysis

Aqueous humor enhances the proliferation of rat retinal precursor cells in culture, and this effect is partially reproduced by ascorbic acid.