Analysis of the correlation between immune cell characteristics and insomnia: a Mendelian randomization study.

Insomnia, recognized as a prevalent sleep disorder, has garnered extensive attention within the realm of public health. Recent studies indicate a close interaction between the immune system and sleep; however, the specific mechanism remains not yet fully understood. Based on the publicly available Genome-Wide Association Study (GWAS) data, we used two-sample Mendelian randomization (MR) analyses to investigate the associations between 731 immune cell traits and insomnia risk. Five MR analysis methods and a comprehensive sensitivity analysis were used to evaluate the reliability of the results. In this study, we identified that 14 immune characteristics among four immune profiles [median fluorescence intensity (MFI), relative cell count (RC), absolute cell count (AC), and morphological parameters (MP)] demonstrated a significant causal association with insomnia. Specifically, eight immune cell characteristics were associated with an increased risk of insomnia, including CD11c+ monocyte% (P < 0.001), CD11c+ HLA DR++ monocyte% (P = 0.004), CD86+ plasmoid dendritic cell (DC) AC (P < 0.001), CD33br HLA DR+ CD14dim AC (P < 0.001), CD8dim AC (P = 0.002), CCR2 on CD14+ CD16- monocyte (P < 0.001), CD39 on monocyte (P < 0.001), and SSC-A on myeloid DC (P < 0.001). Six immune cell characteristics demonstrated protective effects against insomnia, including PB/PC %B cell (P < 0.001), CM CD4+% CD4+ (P < 0.001), T-cell AC (P < 0.001), BAFF-R on IgD- CD38br (P < 0.001), CD16-CD56 on HLA DR+ NK cells (P < 0.001), and CD14 on CD33br HLA DR+ CD14dim (P < 0.001). Our study established the correlation between immune cell characteristics and insomnia, offering a novel theoretical foundation for the concept of sleep-immune cross talk.NEW & NOTEWORTHY This study investigated the association between 731 immune cell characteristics and insomnia using Mendelian randomization, revealing that 14 immune cell characteristics across four groups of immune traits (MFI, RC, AC, and MP) have a significant and causal association with insomnia risk. Our results contribute to the understanding of the sleep-immune cross talk doctrine and offer a new theoretical basis for immune modulation in treating insomnia.

Production and release of 2-MIB in Pseudanabaena: Effects of growth phases on cell characteristics and 2-MIB yield.

2-methylisoborneol (2-MIB), a secondary metabolite produced by cyanobacteria, often causes a musty odour in water, threatening the safety of drinking water supplies. This study investigated the effects of the growth phases on the production of 2-MIB by Pseudanabaena. The effects of cell characteristics on the production and release of 2-MIB were also explored. The total 2-MIB concentration increased during the exponential phase and decreased during the declining phase, which was consistent with the changes in cell density. However, the total 2-MIB yield (1.12-1.27 fg cell-1) of Pseudanabaena did not significantly differ throughout the growth cycle (p > 0.05). Meanwhile, the extracellular 2-MIB yield increased significantly from 0.31 fg cell-1 in the exponential phase to 0.76 fg cell-1 in the declining phase (p < 0.05), and the corresponding proportion of extracellular 2-MIB improved from 25.13% to 59.16% (p < 0.05). The surge in extracellular 2-MIB during the declining phase could be attributed to the breaking of the Pseudanabaena filament, as indicated by the decrease in Dmean during cell ageing. The findings of this study contribute to a more inclusive comprehension and management of musty odour issues resulting from cyanobacteria in the water supply.

Sexual dimorphism in peripheral blood cell characteristics linked to recanalization success of endovascular thrombectomy in acute ischemic stroke.

Endovascular thrombectomy (EVT) success to treat acute ischemic stroke varies with factors like stroke etiology and clot composition, which can differ between sexes. We studied if sex-specific blood cell characteristics (BCCs) are related to recanalization success. We analyzed electronic health records of 333 EVT patients from a single intervention center, and extracted 71 BCCs from the Sapphire flow cytometry analyzer. Through Sparse Partial Least Squares Discriminant Analysis, incorporating cross-validation and stability selection, we identified BCCs associated with successful recanalization (TICI 3) in both sexes. Stroke etiology was considered, while controlling for cardiovascular risk factors. Of the patients, successful recanalization was achieved in 51% of women and 49% of men. 21 of the 71 BCCs showed significant differences between sexes  (pFDR-corrected < 0.05). The female-focused recanalization model had lower error rates than both combined [t(192.4) = 5.9, p < 0.001] and male-only models [t(182.6) = - 15.6, p < 0.001]. In women, successful recanalization and cardioembolism were associated with a higher number of reticulocytes, while unsuccessful recanalization and large artery atherosclerosis (LAA) as cause of stroke were associated with a higher mean corpuscular hemoglobin concentration. In men, unsuccessful recanalization and LAA as cause of stroke were associated with a higher coefficient of variance of lymphocyte complexity of the intracellular structure. Sex-specific BCCs related to recanalization success varied and were linked to stroke etiology. This enhanced understanding may facilitate personalized treatment for acute ischemic stroke.

Identification of stemness in primary retinoblastoma cells by analysis of stem-cell phenotypes and tumorigenicity with culture and xenograft models.

Retinoblastoma (RB) is a primary intraocular malignancy in childhood, and may develop relapse and metastatic disease. This study was to identify the stem-cell properties of primary retinoblastoma cells critical to tumorigenesis and metastasis. Primary cells were isolated from fresh human RB tissues after enucleation, and cultured in serum-free or serum-enriched conditions, with two RB cell-lines Weri-RB1 and Y79 for comparison. Proliferation of primary RB cells were well-maintained in serum-free condition of DMEM/F12 medium, and formed stem-cell like spheroids. The immaturity of cultured primary RB cells was demonstrated by tendency of highly expressed stem-cell markers (CD133, Nestin and OCT4) and suppressed mature retinal-cell markers (GFAP, MAP2 and Recoverin). CD133, a neural stem-cell marker being exclusively studied in RB, was found positive in small patches of cells in archival human RB by immunohistochemistry. Meanwhile, at initial isolation, insignificant CD133+ cells were detected by flow-cytometry, and substantial increase of positivity was observed after several days cultivated in serum-free condition. Cultured primary RB cells were engrafted in subretinal region of BALB/c nude mice for assessment of tumorigenicity. Strong tumorigenic activity and extensive progression of the xenograft retinoblastoma was induced by primary cells as compared with the two cell-lines. Again, immunohistochemistry confirmed that the stem-cell markers were emphasized in the xenograft tumor in mice. Our findings demonstrated that in comparison to the well-established RB cell-lines, cultured primary RB cells possess stem-cell like properties with highly expressed stem-cell markers, self-regenerative growth in culture, and strong in vivo oncogenic potentiality.

Nonionic surfactants induced changes in cell characteristics and phenanthrene degradation ability of Sphingomonas sp. GY2B.

Surfactant-mediated bioremediation has been widely applied in decontaminating PAH-polluted sites. However, the impacts of surfactants on the biodegradation of PAHs have been controversial in the past years. To gain a clear insight into the influencing mechanisms, three nonionic surfactants (Tween80, TritonX-100 and Brij30) were selected to systematically investigate their effects on cell surface properties (membrane permeability, functional groups and elements), cell vitality as well as subsequent phenanthrene degradation ability of Sphingomonas sp. GY2B. Results showed that biodegradation of phenanthrene was stimulated by Tween80, slightly inhibited by TritonX-100 and severely inhibited by Brij30, respectively. Positive effect of Tween80 may arise from its role as the additional carbon source for GY2B to increase bacterial growth and activity, as demonstrated by the increasing viable cells in Tween80 amended degradation systems determined by flow cytometry. Although TritonX-100 could inhibit bacterial growth and disrupt cell membrane, its adverse impacts on microbial cells were weaker than Brij30, which may result in its weaker inhibitive extent. Results from this study can provide a rational basis on selecting surfactants for enhancing bioremediation of PAHs.

Advanced Properties of Urine Derived Stem Cells Compared to Adipose Tissue Derived Stem Cells in Terms of Cell Proliferation, Immune Modulation and Multi Differentiation.

Adipose tissue stem cells (ADSCs) would be an attractive autologous cell source. However, ADSCs require invasive procedures, and has potential complications. Recently, urine stem cells (USCs) have been proposed as an alternative stem cell source. In this study, we compared USCs and ADSCs collected from the same patients on stem cell characteristics and capacity to differentiate into various cell lineages to provide a useful guideline for selecting the appropriate type of cell source for use in clinical application. The urine samples were collected via urethral catheterization, and adipose tissue was obtained from subcutaneous fat tissue during elective laparoscopic kidney surgery from the same patient (n = 10). Both cells were plated for primary culture. Cell proliferation, colony formation, cell surface markers, immune modulation, chromosome stability and multi-lineage differentiation were analyzed for each USCs and ADSCs at cell passage 3, 5, and 7. USCs showed high cell proliferation rate, enhanced colony forming ability, strong positive for stem cell markers expression, high efficiency for inhibition of immune cell activation compared to ADSCs at cell passage 3, 5, and 7. In chromosome stability analysis, both cells showed normal karyotype through all passages. In analysis of multi-lineage capability, USCs showed higher myogenic, neurogenic, and endogenic differentiation rate, and lower osteogenic, adipogenic, and chondrogenic differentiation rate compared to ADSCs. Therefore, we expect that USC can be an alternative autologous stem cell source for muscle, neuron and endothelial tissue reconstruction instead of ADSCs.

Analysis of the stem cell characteristics of adult stem cells from Arbas white Cashmere goat.

Studies have shown that multipotent adult stem cells possess differentiation characteristics similar to embryonic stem cells and pluripotent stem cells. We aimed to explore these similarities further by examining the expression of the pluripotency and stemness biomarkers, AKP, IL-6, Nanog, Oct-4, Rex-1, Sox-2 and TERT, as well as the triploblastic biomarkers, Sox-1, Myod1 and Gata-6 in adipose-derived stem cells (ADSCs), bone marrow stem cells (BMSCs) and muscle-derived satellite cells (MDSCs). These were isolated from adult Arbas white Cashmere goats and cultured in vitro. Immunocytochemistry, reverse transcription quantitative PCR and Western blotting were used to analyze the protein and mRNA expression of the markers. To investigate the ability of ADSCs, BMSCs and MDSCs to differentiate and cause tumors in vivo they were injected into immunodeficient mice (NOD-SCID). All results were compared to those for mouse embryonic stem cells (mESCs). Immunocytochemistry showed that AKP, IL-6, Nanog, Oct-4, Rex-1 and TERT were expressed in ADSCs, BMSCs and MDSCs, whereas Sox-2 was not. In ADSCs, the expression of IL-6 mRNA was relatively high, followed by Nanog and Oct-4, while Rex-1 and TERT expression were the lowest (P<0.01). In BMSCs, the expression of Rex-1 was relatively high, followed by IL-6, while Oct-4, Nanog and TERT were comparatively low (P<0.01). In MDSCs, the expression of IL-6, Nanog and Oct-4 were relatively high, while TERT was comparatively low (P<0.01). However, no expression of Sox-2 mRNA was detected in any of the three cell lines. The expression of Sox-1, Myod1 and Gata-6 was observed to different degrees in all three cell lines (P<0.01); the expression pattern in MDSCs was different from that in ADSCs and BMSCs. Western blotting indicated that no expression of Sox-2 and Rex-1 protein occurred in ADSCs, BMSCs and MDSCs, while the other five proteins were all expressed to different degrees (P<0.01); the expression pattern was consistent with the mRNA results. In contrast to the mESCs, no teratoma tissue or triploblastic differentiation appendages were formed in the immunodeficient mice after injection of ADSCs, BMSCs and MDSCs. Our results suggest that the three adult goat stem cell types are non-oncogenic and have stemness characteristics similar to embryonic stem cells. Of these, MDSCs were found to exhibit the most ESC-like properties and would make the best candidates for clinical application.

Optimization of stage conversion time and modification of cell metabolism to enhance lipid production of Auxenochlorella pyrenoidosa in two-stage cultivation.

Traditionally, the time of maximum biomass concentration in stage I is the widely adopted stage conversion time in two-stage microalgae culture. This study challenges this conventional approach, demonstrating that the optimal stage conversion time in stage I is 72 h rather than 120 h for achieving maximum biomass concentration. A comparison of cell characteristics revealed that algal cells at 72 h exhibited better growth potential, leading to a higher biomass concentration after transfer to stage II and, consequently, increased lipid productivity. Moreover, the use of phosphorus repletion (5-fold) in stage II directed carbon flux toward biomass growth and lipid accumulation, thereby enhancing lipid productivity. By optimizing the stage conversion time and implementing phosphorus repletion, the mean lipid productivity of Auxenochlorella pyrenoidosa cultured under autotrophy-nitrogen starvation and autotrophy-high light conditions increased by 31 % and 60 %, respectively. This study underscores the importance of reevaluating the currently widely used stage conversion time.

Brain pericyte biology: from physiopathological mechanisms to potential therapeutic applications in ischemic stroke.

Pericytes play an indispensable role in various organs and biological processes, such as promoting angiogenesis, regulating microvascular blood flow, and participating in immune responses. Therefore, in this review, we will first introduce the discovery and development of pericytes, identification methods and functional characteristics, then focus on brain pericytes, on the one hand, to summarize the functions of brain pericytes under physiological conditions, mainly discussing from the aspects of stem cell characteristics, contractile characteristics and paracrine characteristics; on the other hand, to summarize the role of brain pericytes under pathological conditions, mainly taking ischemic stroke as an example. Finally, we will discuss and analyze the application and development of pericytes as therapeutic targets, providing the research basis and direction for future microvascular diseases, especially ischemic stroke treatment.

Human bone marrow stromal cells: the impact of anticoagulants on stem cell properties.

Background: Bone marrow stromal cells (BMSCs) are the source of multipotent stem cells, which are important for regenerative medicine and diagnostic purposes. The isolation of human BMSCs from the bone marrow (BM) cavity using BM aspiration applies the method with collection into tubes containing anticoagulants. Interactions with anticoagulants may affect the characteristics and composition of isolated BMSCs in the culture. Thus, we investigated how anticoagulants in isolation procedures and cultivation affect BMSC molecular characteristics. Methods: BM donors (age: 48-85 years) were recruited from the hematology clinic. BM aspirates were obtained from the iliac crest and divided into tubes coated with ethylenediaminetetraacetic acid (EDTA) or heparin anticoagulants. Isolated BMSCs were analyzed by flow cytometry and RNA-seq analysis. Further cellular and molecular characterizations of BMSCs including CFU, proliferation and differentiation assays, cytometry, bioenergetic assays, metabolomics, immunostaining, and RT-qPCR were performed. Results: The paired samples of isolated BMSCs obtained from the same patient showed increased cellular yield in heparin vs. EDTA samples, accompanied by the increased number of CFU colonies. However, no significant changes in molecular characteristics were found between heparin- and EDTA-isolated BMSCs. On the other hand, RNA-seq analysis revealed an increased expression of genes involved in nucleotide metabolism and cellular metabolism in cultivated vs. non-cultivated BMSCs regardless of the anticoagulant, while genes involved in inflammation and chromatin remodeling were decreased in cultivated vs. non-cultivated BMSCs. Conclusion: The type of anticoagulant in BMSC isolation did not have a significant impact on molecular characteristics and cellular composition, while in vitro cultivation caused the major change in the transcriptomics of BMSCs, which is important for future protocols using BMSCs in regenerative medicine and clinics.

Characteristics of peripheral blood cells are independently related to major adverse cardiovascular events after carotid endarterectomy.

Background and aims: Patients who underwent carotid endarterectomy (CEA) still have a residual risk of 13% of developing a major adverse cardiovascular event (MACE) within 3 years. Inflammatory processes leading up to MACE are not fully understood. Therefore, we examined blood cell characteristics (BCCs), possibly reflecting inflammatory processes, in relation to MACE to identify BCCs that may contribute to an increased risk. Methods: We analyzed 75 pretreatment BCCs from the Sapphire analyzer, and clinical data from the Athero-Express biobank in relation to MACE after CEA using Random Survival Forests, and a Generalized Additive Survival Model. To understand biological mechanisms, we related the identified variables to intraplaque hemorrhage (IPH). Results: Of 783 patients, 97 (12%) developed MACE within 3 years after CEA. Red blood cell distribution width (RDW) (HR 1.23 [1.02, 1.68], p = 0.022), CV of lymphocyte size (LACV) (HR 0.78 [0.63, 0.99], p = 0.043), neutrophil complexity of the intracellular structure (NIMN) (HR 0.80 [0.64, 0.98], p = 0.033), mean neutrophil size (NAMN) (HR 0.67 [0.55, 0.83], p < 0.001), mean corpuscular volume (MCV) (HR 1.35 [1.09, 1.66], p = 0.005), eGFR (HR 0.65 [0.52, 0.80], p < 0.001); and HDL-cholesterol (HR 0.62 [0.45, 0.85], p = 0.003) were related to MACE. NAMN was related to IPH (OR 0.83 [0.71-0.98], p = 0.02). Conclusions: This is the first study to present a higher RDW and MCV and lower LACV, NIMN and NAMN as biomarkers reflecting inflammatory processes that may contribute to an increased risk of MACE after CEA.

Role of PrPC in Cancer Stem Cell Characteristics and Drug Resistance in Colon Cancer Cells.

BACKGROUND/AIM: Cancer stem cell characteristics and drug resistance of colorectal cancer are associated with failure of cancer treatment. In this study, we investigated the effects of PrPC on cancer stem cell characteristics, migration, invasion, and drug resistance of 5FU-resistant CRC cells. MATERIALS AND METHODS: PrPC negative and PrPC positive cells were isolated from 5FU-resistant CRC cells using magnetic activated cell sorting. Sphere formation, cancer stem cell marker expression, migration, invasion, and drug resistance were analyzed. RESULTS: PrPC positive cells showed increased sphere formation capacity and increased expression of cancer stem cell markers compared to PrPC negative cells. In addition, PrPC positive cells showed increased migration, invasion and drug resistance compared to PrPC negative cells. Furthermore, knockdown of PrPC abolished these effects. CONCLUSION: PrPC expression is important in CRC cell behavior, such as sphere formation, migration, invasion, and drug resistance. PrPC is an important therapeutic target for the treatment of CRC.

Growth and Stem Cell Characteristics of Tendon-Derived Cells with Different Initial Seeding Densities: An In Vitro Study in Mouse Flexor Tendon Cells.

Tendon stem/progenitor cells (TSPCs) are considered promising seed cells for tendon regeneration. Previous studies reported that a low seeding density favors TSPC growth, whereas a high seeding density favors tenocyte growth. We aimed to distinguish TSPCs from tenocytes by seeding tendon-derived cells at a density gradient. In this study, tendon-derived cells were isolated from flexor digitorum profundus tendons of mice and seeded at the initial densities of 50, 500, 5,000, and 50,000/cm2. We found that distinct cell colonies were formed from cells with initial seeding densities of 50 and 500/cm2, but colonies were not discernible for cells seeded at 5,000 and 50,000/cm2. There was a positive correlation between cell proliferation rate and seeding density, but a negative correlation between cell senescence and seeding density. The cell proliferation rate decreased gradually during serial passages. All cells exhibited restricted differentiation potentials, and expressed stem cell markers and relatively high levels of tenogenic markers without notable differences among cells seeded at different densities. We concluded that a pure population of TSPCs could not be isolated from mouse digital flexor tendons through culturing cells at a density gradient. Cells seeded at low densities had very limited proliferative ability and did not show more prominent stem cell characteristics when compared with cells seeded at high densities.

A Rising Star in Pancreatic Diseases: Pancreatic Stellate Cells.

Pancreatic stellate cell (PSC) is a type of pluripotent cell located between pancreatic lobules and the surrounding area of acinars. When activated, PSC can be transformed into myofibroblast-like cell. A number of evidences suggest that activated PSC is the main source of the accumulation of extracellular matrix (ECM) protein under the pathological conditions, which lead to pancreatic fibrosis in chronic pancreatitis and pancreatic cancer. Recent studies have found that PSC also plays an important role in the endocrine cell function, islet fibrosis and diabetes. In order to provide new strategies for the treatment of pancreatic diseases, this paper systematically summarizes the recent researches about the biological behaviors of PSC, including its stem/progenitor cell characteristics, secreted exosomes, cellular senescence, epithelial mesenchymal transformation (EMT), energy metabolism and direct mechanical reprogramming.

Simultaneous Cr(VI) removal and 2,2',4,4'-tetrabromodiphenyl ether (BDE-47) biodegradation by Pseudomonas aeruginosa in liquid medium.

Simultaneous Cr(VI) removal and 2,2',4,4'-tetra brominated diphenyl ether (BDE-47) biodegradation by Pseudomonas aeruginosa in liquid medium were investigated in this study, with the goal of elucidating the interaction between concomitant pollutants Cr(VI) and BDE-47 during microbial remediation. The experimental results revealed that the degradation efficiency of 1 mg L(-1) BDE-47 by 60 mg L(-1) biomass achieved 51.3% within 7 d when 2 mg L(-1) Cr(VI) coexisted. The degradation efficiency was accelerated at low concentrations of Cr(VI) (≤5 mg L(-1)), but inhibited at higher levels (≥10 mg L(-1)). Cr(VI) of 2 mg L(-1) facilitated the secretion of rhamnolipid from the strain, altered cell surface hydrophobicity and cell membrane permeability, and promoted intracellular BDE-47 accumulation, thus improving BDE-47 biotransformation. In addition, the stimulation of intracellular enzyme synthesis by 2 mg L(-1) Cr(VI) contributed to more BDE-47 elimination in the cells. The achievement of BDE-47 biodegradation was coupled with cell growth, enzyme extraction, cell membrane permeability change, and ATPase activity increase. The study also indicated that the improvement of Cr(VI) removal in BDE-47/Cr(VI) co-contaminated condition was mostly due to the increasing synthesis of extracellular enzyme in the presence of low concentrations of BDE-47. The whole study demonstrated that P. aeruginosa was available for the removal of toxic Cr(VI) and degradation of BDE-47 simultaneously in the liquid.

Stem cell properties and neural differentiation of sheep amniotic epithelial cells.

This study was designed to verify the stem cell properties of sheep amniotic epithelial cells and their capacity for neural differentiation. Immunofluorescence microscopy and reverse transcription-PCR revealed that the sheep amniotic epithelial cells were positive for the embryonic stem cell marker proteins SSEA-1, SSEA-3, SSEA-4, TRA-1-60 and TRA-1-81, and the totipotency-associated genes Oct-4, Sox-2 and Rex-1, but negative for Nanog. Amniotic epithelial cells expressed ß-III-tubulin, glial fibrillary acidic protein, nestin and microtubule-associated protein-2 at 28 days after induction with serum-free neurobasal-A medium containing B-27. Thus, sheep amniotic epithelial cells could differentiate into neurons expressing ß-III-tubulin and microtubule-associated protein-2, and glial-like cells expressing glial fibrillary acidic protein, under specific conditions.

Do the Fibroblasts Contained in Early Passage MSC Population Adversely Affect the Characteristics of Stem Cell Population Obtained from Human Placenta?

BACKGROUND AND OBJECTIVES: Mesenchymal stem cells (MSCs) have been obtained from various human tissues by harvesting plastic adherent fibroblast-like cell population. For potential use in regeneration medicine, early passage MSC population is preferred to avoid cell senescence. The early passage adherent cell population contains MSCs as well as fibroblasts, however, the significance of the contained fibroblasts has not been well investigated. Thus, we investigated the stem cell characteristics of the early passage MSC population with and without fibroblasts depletion. METHODS AND RESULTS: We obtained adherent cell populations from full term placenta at passage 2∼3 and divided them into two subpopulations: fibroblasts depleted (popFD) and non-depleted population (popFND) using magnetic cell sorting method. The two subpopulations were compared in terms of cell morphology, potential for long term culture, colony forming ability, and tri-lineage differentiation for adipogenic, chondrogenic, and osteogenic differentiation. The percentage of fibroblasts contained in the early passage MSC population was 5.3% (2.9∼8.4). Both the popFD and popFND was spindle shaped from early passages and maintained long term culture up to 20∼22 passages. CFU-F assay showed no difference between the subpopulations. Overall, tri-lineage differentiation showed a tendency of better differentiation potential of popFND than popFD. CONCLUSIONS: We confirmed that fibroblasts are contained in early population of placenta-derived MSCs obtained by current method. This study revealed that the contained fibroblasts in early passage MSC population do not adversely affect the properties of MSCs in terms of cell morphology, potential for long term culture, colony forming ability, and tri-lineage differentiation.

Advanced Properties of Urine Derived Stem Cells Compared to Adipose Tissue Derived Stem Cells in Terms of Cell Proliferation, Immune Modulation and Multi Differentiation

Adipose tissue stem cells (ADSCs) would be an attractive autologous cell source. However, ADSCs require invasive procedures, and has potential complications. Recently, urine stem cells (USCs) have been proposed as an alternative stem cell source. In this study, we compared USCs and ADSCs collected from the same patients on stem cell characteristics and capacity to differentiate into various cell lineages to provide a useful guideline for selecting the appropriate type of cell source for use in clinical application. The urine samples were collected via urethral catheterization, and adipose tissue was obtained from subcutaneous fat tissue during elective laparoscopic kidney surgery from the same patient (n = 10). Both cells were plated for primary culture. Cell proliferation, colony formation, cell surface markers, immune modulation, chromosome stability and multi-lineage differentiation were analyzed for each USCs and ADSCs at cell passage 3, 5, and 7. USCs showed high cell proliferation rate, enhanced colony forming ability, strong positive for stem cell markers expression, high efficiency for inhibition of immune cell activation compared to ADSCs at cell passage 3, 5, and 7. In chromosome stability analysis, both cells showed normal karyotype through all passages. In analysis of multi-lineage capability, USCs showed higher myogenic, neurogenic, and endogenic differentiation rate, and lower osteogenic, adipogenic, and chondrogenic differentiation rate compared to ADSCs. Therefore, we expect that USC can be an alternative autologous stem cell source for muscle, neuron and endothelial tissue reconstruction instead of ADSCs.

Do the Fibroblasts Contained in Early Passage MSC Population Adversely Affect the Characteristics of Stem Cell Population Obtained from Human Placenta?

BACKGROUND AND OBJECTIVES: Mesenchymal stem cells (MSCs) have been obtained from various human tissues by harvesting plastic adherent fibroblast-like cell population. For potential use in regeneration medicine, early passage MSC population is preferred to avoid cell senescence. The early passage adherent cell population contains MSCs as well as fibroblasts, however, the significance of the contained fibroblasts has not been well investigated. Thus, we investigated the stem cell characteristics of the early passage MSC population with and without fibroblasts depletion. METHODS AND RESULTS: We obtained adherent cell populations from full term placenta at passage 2~3 and divided them into two subpopulations: fibroblasts depleted (popFD) and non-depleted population (popFND) using magnetic cell sorting method. The two subpopulations were compared in terms of cell morphology, potential for long term culture, colony forming ability, and tri-lineage differentiation for adipogenic, chondrogenic, and osteogenic differentiation. The percentage of fibroblasts contained in the early passage MSC population was 5.3% (2.9~8.4). Both the popFD and popFND was spindle shaped from early passages and maintained long term culture up to 20~22 passages. CFU-F assay showed no difference between the subpopulations. Overall, tri-lineage differentiation showed a tendency of better differentiation potential of popFND than popFD. CONCLUSIONS: We confirmed that fibroblasts are contained in early population of placenta-derived MSCs obtained by current method. This study revealed that the contained fibroblasts in early passage MSC population do not adversely affect the properties of MSCs in terms of cell morphology, potential for long term culture, colony forming ability, and tri-lineage differentiation.