Current status and challenges for cell-cultured milk technology: a systematic review.

Cellular agriculture is an innovative technology for manufacturing sustainable agricultural products as an alternative to traditional agriculture. While most cellular agriculture is predominantly centered on the production of cultured meat, there is a growing demand for an understanding of the production techniques involved in dairy products within cellular agriculture. This review focuses on the current status of cellular agriculture in the dairy sector and technical challenges for cell-cultured milk production. Cellular agriculture technology in the dairy sector has been classified into fermentation-based and animal cell culture-based cellular agriculture. Currently, various companies synthesize milk components through precision fermentation technology. Nevertheless, several startup companies are pursuing animal cell-based technology, driven by public concerns regarding genetically modified organisms in precision fermentation technology. Hence, this review offers an up-to-date exploration of animal cell-based cellular agriculture to produce milk components, specifically emphasizing the structural, functional, and productive aspects of mammary epithelial cells, providing new information for industry and academia.

The Role of Progesterone in Elf5 Activation and Milk Component Synthesis for Cell-Cultured Milk Production in MAC-T Cells.

Prolactin is essential for mammary gland development and lactation. Progesterone also induces ductal branching and alveolar formation via initial secretory differentiation within the mammary gland. Herein, we aimed to evaluate the role of progesterone as a prolactin substitute for the production of cell-cultured milk components in MAC-T cells. Cells were treated with various hormones such as prolactin (PRL), progesterone (P4), 17ß-estradiol (E2), cortisol (COR), and insulin (INS) for 5 d. MAC-T cells cultured in a P4 differentiation media (2500 ng/mL of P4, 25 ng/mL of E2, 25 ng/mL of COR, and 25 ng/mL of INS) showed similar levels of E74-like factor 5 (Elf5) and milk component synthesis (α-casein, ß-casein, α-lactalbumin, ß-lactoglobulin, and triglycerides) compared to those cultured in a PRL differentiation media (5000 ng/mL of PRL, 500 ng/mL of CORT, and 50 ng/mL of INS). The levels of α-casein and triglycerides in the optimal P4 differentiation media were present at comparable levels to those in the PRL differentiation media. Our results demonstrated that P4 induces the activation of Elf5 and the synthesis of milk components in MAC-T cells, similar to PRL. Therefore, P4 may be used as an effective substitute of PRL for cell-cultured milk production in in vitro frameworks.

Optimizing hormonal and amino acid combinations for enhanced cell proliferation and cell cycle progression in bovine mammary epithelial cells.

OBJECTIVE: The number of bovine mammary epithelial cells (BMECs) is closely associated with the quantity of milk production in dairy cows; however, the optimal levels and the combined effects of hormones and essential amino acids (EAAs) on cell proliferation are not completely understood. Thus, the purpose of this study was to determine the optimal combination of individual hormones and EAAs for cell proliferation and related signaling pathways in BMECs. METHODS: Immortalized BMECs (MAC-T) were treated with six hormones (insulin, cortisol, progesterone, estrone, 17ß-estradiol, and epidermal growth factor) and ten EAAs (arginine, histidine, leucine, isoleucine, threonine, tryptophan, lysine, methionine, phenylalanine, and valine) for 24 h. RESULTS: Cells were cultured in a medium containing 10% fetal bovine serum (FBS) as FBS supplemented at a concentration of 10% to 50% showed a comparable increase in cell proliferation rate. The optimized combination of four hormones (insulin, cortisol, progesterone, and 17ß-estradiol) and 20% of a mixture of ten EAAs led to the highest cell proliferation rate, which led to a significant increase in cell cycle progression at the S and G2/M phases, in the protein levels of proliferating cell nuclear antigen and cyclin B1, cell nucleus staining, and in cell numbers. CONCLUSION: The optimal combination of hormones and EAAs increased BMEC proliferation by enhancing cell cycle progression in the S and G/2M phases. Our findings indicate that optimizing hormone and amino acid levels has the potential to enhance milk production, both in cell culture settings by promoting increased cell numbers, and in dairy cows by regulating feed intake.

Development of ß-Cyclodextrin/Konjac-Based Emulsion Gel for a Pork Backfat Substitute in Emulsion-Type Sausage.

Emulsion gel has been used to replace animal fats in meat products. Konjac is a widely used gelling agent; however, its low emulsion stability limits its use in meat products. This study aimed to examine the quality characteristics of ß-cyclodextrin (CD)-supplemented konjac-based emulsion gel (KEG) (CD-KEG) and its application as a fat substitute in emulsion-type sausages. The supplementation of CD increased hydrogen bonds and hydrophobic interactions with konjac and oil in the gels, respectively. Additionally, CD increased the structural complexity and strength of KEG. Since adding more than 6% of CD to KEG did not increase the gel strength, 6% CD-added KEG was adopted to substitute for pork backfat in manufacturing low-fat emulsion-type sausages. The following formulations of the sausages were prepared: pork backfat 20% (PF20); pork backfat 10% + KEG 10% (KEG10); KEG 20% (KEG20); pork backfat 10% + CD-KEG 10% (CD-KEG10); CD-KEG 20% (CD-KEG20); and pork backfat 5% (PF5). The CD-KEG20 formulation exhibited higher viscosity and viscoelasticity than KEG20, which suggested that CD improves the rheological properties and the thermal stability of meat batter. Additionally, CD-KEG20 showed similar emulsion stability, cooking yield and texture parameters compared with PF20. Therefore, 6% CD-added KEG is a suitable fat substitute for preparing low-fat emulsion-type sausages.

Effects of duck fat and κ-carrageenan as replacements for beef fat and pork backfat in frankfurters.

OBJECTIVE: Frankfurters are emulsion-type sausages that are widely consumed worldwide. However, some concerns regarding negative health effects have been raised because of the high fat content and the type of fat. This study aimed to evaluate the effects of duck fat and κ-carrageenan as replacements for beef fat and pork backfat in frankfurters. METHODS: The different formulations for the frankfurters were as follows: 20% beef fat (BF), 20% pork backfat (PBF), 20% duck fat (DF), 20% soybean oil (SO), 20% duck fat/1% κ-carrageenan (DFC), and 20% soybean oil/1% κ-carrageenan (SOC). Physicochemical (fatty acid profile, color, rheological properties, cooking loss, water holding capacity, emulsion stability, and texture profile analysis), oxidative stability and sensory properties of frankfurters were evaluated. RESULTS: Duck fat and κ-carrageenan improved rheological properties of meat batter, and physicochemical properties (emulsion stability, cooking loss, and hardness) of frankfurters. Moreover, duck fat added-frankfurters (DF and DFC) had higher oxidative stability than that of soybean-added frankfurters (SO and SOC) during refrigerated storage for 28 days. In sensory evaluation, flavor, texture, and overall acceptability of DFC were acceptable to untrained panelists. CONCLUSION: Our data suggest that duck fat and κ-carrageenan can replace beef fat and pork backfat in frankfurters. Duck fat and κ-carrageenan contributed to improve the physicochemical properties and oxidative stability while maintaining sensory properties. Therefore, the use of duck fat and κ-carrageenan may be a suitable alternative for replacing beef fat or pork backfat in frankfurters.

Toxicity Assessment of a Single Dose of Poly(ethylene glycol) Diglycidyl Ether (PEGDE) Administered Subcutaneously in Mice.

Poly(ethylene glycol) diglycidyl ether (PEGDE) is widely used to cross-link polymers, particularly in the pharmaceutical and biomaterial sectors. However, the subcutaneous toxicity of PEGDE has not yet been assessed. PEGDE samples (500-40,000 µg/mouse) were subcutaneously injected into the paraspinal dorsum of BALB/c male mice. Cage-side observations were carried out with measurement of organ weight, body weight variation, and feed intake, as well as histopathological characterization on day 28 post-exposure. Mice that received 40,000 µg of PEGDE showed severe toxic response and had to be euthanized. Subcutaneous injection of PEGDE did not alter feed intake and organ weight; however, the body weight variation of mice injected with 20,000 µg of PEGDE was significantly lower than that of the other groups. Exposure to 10,000 and 20,000 µg of PEGDE induced epidermal ulcer formation and hair loss. The histology of skin tissue in mice administered with 20,000 µg of PEGDE showed re-epithelialized or unhealed wounds. However, the liver, spleen, and kidneys were histologically normal. Collectively, PEGDE, particularly above 10,000 µg/mouse, caused subcutaneous toxicity with ulceration, but no toxicity in the other organs. These results may indicate the optimal concentration of subcutaneously injected PEGDE.

Early experience with a thermo-expandable stent (memokath) for the management of recurrent urethral stricture.

PURPOSE: To report our early experience with thermo-expandable urethral stents (Memokath) for the management of recurrent urethral stricture and to assess the efficacy of urethral stents. MATERIALS AND METHODS: Between March 2012 and February 2013, 13 patients with recurrent urethral stricture after several attempts with direct visual internal urethrotomy (DVIU) or failed urethroplasty underwent DVIU with thermally expandable, nickel-titanium alloy urethral stent (Memokath) insertion. Follow-up study time points were at 1, 3, 6, 9, and 12 months after stent insertion. Follow-up evaluation included uroflowmetry, retrograde urethrogram, plain radiography, and urinalysis. RESULTS: The mean patient age was 47.7 years (range, 18 to 74 years). The mean urethral stricture length was 5.54 cm (range, 1 to 12 cm). There were six patients with bulbar, four patients with proximal penile, one patient with distal penile, and two patients with whole penile urethral strictures, respectively. The overall success rate was 69% (9/13) and the mean postoperative peak flow rate was 17.7 mL/s (range, 6 to 28 mL/s). Major complications occurred in four patients including one patient (7.7%) with urethrocutaneous fistula induced by the stent and three patients with urethral hyperplasia. The mean follow-up duration was 8.4 months. CONCLUSIONS: Our initial clinical experience indicates that thermo-expandable stents can be another temporary management option for recurrent urethral stricture patients who are unfit for or refuse urethroplasty. Distal or whole penile urethral stricture can be factors predicting poor results.

Solitary testicular metastasis of prostate cancer mimicking primary testicular cancer.

We report a rare case of testicular metastasis from prostate cancer. A 68-year-old patient presented with a right testicular mass with discomfort. He had a history of robot-assisted laparoscopic radical prostatectomy and had received adjuvant radiation therapy and had been treated with androgen deprivation therapy continuously at another institution. We performed a right inguinal orchiectomy. The testicular mass was diagnosed as a metastasis from prostate carcinoma.