scholarly journals Endometrial Mesenchymal Stem Cells Isolated from Menstrual Blood by Adherence

2016 ◽  
Vol 2016 ◽  
pp. 1-8 ◽  
Author(s):  
Xue Du ◽  
Qing Yuan ◽  
Ye Qu ◽  
Yuan Zhou ◽  
Jia Bei
Keyword(s):  
Growth Factor ◽  
Alcian Blue ◽  
Gradient Centrifugation ◽  
Menstrual Blood ◽  
Blue Staining ◽  
Enzyme Linked Immunosorbent Assay ◽  
Alcian Blue Staining ◽  
Promising Alternative ◽  
Oil Red O Staining ◽  
Oil Red O

Objective. To find a convenient and efficient way to isolate MSCs from human menstrual blood and to investigate their biological characteristics, proliferative capacity, and secretion levels.Methods. MSCs were isolated from menstrual blood of 3 healthy women using adherence. Cell immunological phenotype was examined by flow cytometry; the adipogenic, osteogenic, and chondrogenic differentiation of MSCs was examined by Oil-Red-O staining, ALP staining, and Alcian Blue staining, respectively; and the secretion of cytokines, including vascular endothelial growth factor (VEGF), hepatocyte growth factor (HGF), and insulin-like growth factor-1 (IGF-1), was detected using enzyme-linked immunosorbent assay.Results. MB-MSCs were successfully isolated from human menstrual blood using adherence. They were positive for CD73, CD105, CD29, and CD44, but negative for CD31 and CD45. The differentiated MB-MSCs were positive for ALP staining, Oil-Red-O staining, and Alcian Blue staining. In addition, they could secrete antiapoptotic cytokines, such as VEGF, IGF-1, and HGF.Conclusion. It is feasible to isolate MSCs from human menstrual blood, thus avoiding invasive procedures and ethical controversies. Adherence could be a promising alternative to the density gradient centrifugation for the isolation of MSCs from menstrual blood.

210 COMPARATIVE CHARACTERIZATION OF MESENCHYMAL STEM CELLS DERIVED FROM MINIATURE PIG SYNOVIUM, SYNOVIAL FLUID AND BONE MARROW

2012 ◽  
Vol 24 (1) ◽  
pp. 217 ◽  
Author(s):  
W. J. Lee ◽  
G. H. Maeng ◽  
R. H. Jeon ◽  
G. J. Rho ◽  
S. L. Lee
Keyword(s):  
Stem Cells ◽  
Bone Marrow ◽  
Blue Staining ◽  
Alcian Blue Staining ◽  
Miniature Pig ◽  
Rt Pcr ◽  
Alizarin Red ◽  
Differentiated Cells ◽  
Oil Red O Staining ◽  
Oil Red O

Mesenchymal stem cells (MSC) are a valuable cell source for cartilage tissue engineering because MSC derived from various tissues readily differentiate into skeletal cell and chondrogenic lineages. In this study, we compared the cellular characteristics of synovium (SN) and synovial fluid (SF)-derived MSC with bone marrow (BM)-derived MSC of miniature pig. Biopsies of SN and BM were collected and SF was flushed with saline solution from 6-month-old miniature pig (T-type, PWG Micro-pig, PWG Genetics Korea, South Korea). Cells were isolated from collected tissues and cultured in advanced-DMEM supplemented with 10% fetal bovine serum at 38.5°C in a humidified atmosphere of 5% CO2 in air. The cells were then evaluated for their expression of MSC-specific markers, including CD29, CD44 and CD90 using flow cytometry. The expression of early transcriptional factors, such as Oct3/4, Nanog and Sox2 was evaluated by immunocytochemical staining and reverse transcription-PCR (RT-PCR). Multilineage differentiation of MSC were induced under conditions conductive for osteogenic, adipogenic and chondrogenic lineages and then evaluated by von Kossa and Alizarin Red S staining, Oil red O staining and Alcian Blue staining, respectively. Differentiated cells were further analysed for the expression of lineage specific markers by RT-PCR. Statistical analysis was performed using one-way ANOVA by SPSS. The SN-, SF- and BM-MSC were observed to be positive for MSC specific markers, such as CD29 (99 ± 0.2, 96 ± 0.5 and 98 ± 0.2, respectively), CD44 (97 ± 0.3, 96 ± 0.6 and 98 ± 0.5, respectively) and CD90 (95 ± 0.5, 92 ± 0.2 and 96 ± 0.3, respectively); however, haematopoietic marker CD45 (2.0 ± 2.1, 3.0 ± 1.3 and 2.0 ± 3.0, respectively) was barely detected. In all MSC, early transcription factors (Oct3/4, Nanog and Sox2) were expressed by immnocytochemical staining and the transcripts were detected by RT-PCR. Following exposure to the specific differentiation medium, all these MSC were capable of differentiating into mesenchymal lineages, such as osteogenic, adiopogenic and chondrogenic as assessed by von Kossa and Alizarin Red S staining, Oil red O staining and and Alcian Blue staining, respectively. In addition, differentiated cells from all MSC expressed the marker genes specific to osteocytes (osteonectin, Runx2), adipocytes (aP2, PRAR-γ2) and chondrocytes (aggrecan, collagen type 2) by RT-PCR. The results of this study suggested that cells isolated from miniature pig articular tissues, such as SN and SF have similar characteristics of MSCs and their differentiation ability was comparable to BM-MSC. Hence, it is possible to establish MSCs from SN and SF as alternate sources during the biopsy of synovial tissues for arthritis diagnosis. Further studies are being carried out to evaluate their in vivo differentiation potential into chondrocytes. This study was supported by Rural Development Administration (grant No. 20110701-305-533-001-02-00) and National Research Foundation of Korea (grant No. 2011-0010252) of the Republic of Korea.

scholarly journals LARYNGOTRACHEITIS VIRUS IN CHICKENS

1967 ◽  
Vol 125 (3) ◽  
pp. 409-428 ◽  
Author(s):  
Betsy G. Bang ◽  
Frederik B. Bang
Keyword(s):  
Alcian Blue ◽  
Plasma Cells ◽  
Giant Cells ◽  
Functional Restoration ◽  
Nasal Fossa ◽  
Blue Staining ◽  
Alcian Blue Staining ◽  
Cell Nuclei ◽  
Histochemical Change ◽  
Laryngotracheitis Virus

Infectious laryngotracheitis can be produced in chickens as an experimental model of severe nonfatal rhinitis and sinusitis. Inoculated intranasally into unanesthetized baby chicks it remains limited to the nasal fossa, produces acute desquamation of all nasal epithelia, results in functional recovery of the respiratory epithelium, but leaves important residual abnormalities. From the earliest recognizable lesions through 4½ months' convalescence, the principal changes are as follows: 1. Initial lesions, or small syncytia of intranuclear "inclusions", first identifiable in the mucociliated cells of the shallowest portion of the epithelium at about 21 hr postinoculum (the inner surface of the maxillary conchal scroll). 2. Acute sloughing, (about 3 to 7 days), marked by: (a) spread of lesions from cell to cell via multinucleated "giant cells" which progressively slough and desquamate respiratory, olfactory, and sinus epithelia, epithelial neural elements and blood vessels; (b) appearance of numbers of eosinophilic leukocytes along the basement membrane at the sites of lesions just previous to sloughing; intensive infiltration of the submucosa with small lymphocytes after sloughing begins; (c) histochemical change in the intracellular mucus of the cells which comprise the syncytia: this mucus stains with Alcian blue alone when stained with AB-PAS; and (d) all cartilages of the maxillary conchae become flaccid, and the cell nuclei and matrix lose both basophilic and Alcian blue staining properties, effects which recede by about the 8th day. 3. Repair (about 8 to 21 days), marked by rapid initial spread of a sheet of epithelial cells over the infiltrated subrmucosa, appearance of numbers of plasma cells circulating in the tissues, formation of encapsulated secondary nodules, and mucosal adhesions. 4. Convalescence (about 1 to 4½ months when experiments terminated), marked by functional restoration of the mucociliary lining of the nasal fossa. However, at 4½ months eight specimens all show complete metaplasia of the olfactory organ (end nerves, supporting cells, and glands of Bowman) to mucociliated epithelium, all show abnormal formation and alignment of mucous acini, and about 50% have severe persistent sinusitis.

Histochemical observations on the tegumentary epithelium and interproglottidal glands of Moniezia expansa (Rud., 1805) (Cestoda, Cyclophyllidea)

Parasitology ◽  
1969 ◽  
Vol 59 (3) ◽  
pp. 505-518 ◽  
Author(s):  
R. E. Howells ◽  
D. A. Erasmus
Keyword(s):  
Alcian Blue ◽  
Regional Differences ◽  
Succinic Dehydrogenase ◽  
Neck Region ◽  
Secretory Activity ◽  
Blue Staining ◽  
Alcian Blue Staining ◽  
Light Microscope Level ◽  
Gland Cells ◽  
Moniezia Expansa

Regional differences in the tegumentary tissue of Moniezia expansa, as revealed at the light-microscope level by histological and histochemical techniques, are described and evidence for secretory activity by the interproglottidal glands is presented.In very immature proglottides the interproglottidal glands are at the ‘precryptic’ stage. Gland cells may be differentiated from other tegumentary cells by their high RNA content and in certain gland cells the presence of an alcian blue staining material.In mature proglottides the glands consist of rosette-like clusters of cells around crypt-like intuckings of the tegument. Two types of cells are found in the gland, small alcian blue-staining cells which are most numerous in the neck region of the crypt, and larger cells, the predominant gland cells, which do not stain with alcian blue but possess non-specific esterase activity. No other tegumentary cells in Moniezia exhibit this activity. Esterase and phosphatase activity is found in the tegument and crypt of the glands and in the interproglottidal folds.The non-enzyme histochemistry confirms and extends the observations of previous workers.Cytochrome oxidase and succinic dehydrogenase were detected in the tegumentary cells and tegument. Very strong reactions were given in the neck and scolex, with a progressive diminution of activity posteriorly along the strobila. Very low activities were recorded in the tegument of the glands.

Dual action of sonic hedgehog on chondrocyte hypertrophy:retrovirus mediated ectopic sonic hedgehog expression in limb bud micromass culture induces novel cartilage nodules that are positive for alkaline phosphatase and type X collagen

1997 ◽  
Vol 110 (21) ◽  
pp. 2691-2701 ◽  
Author(s):  
N.S. Stott ◽  
C.M. Chuong
Keyword(s):  
Alkaline Phosphatase ◽  
Gene Family ◽  
Alcian Blue ◽  
Sonic Hedgehog ◽  
Ectopic Expression ◽  
Limb Bud ◽  
Blue Staining ◽  
Alcian Blue Staining ◽  
Type X Collagen ◽  
Chondrocyte Maturation

Members of the vertebrate hedgehog gene family (HH) are involved in patterning and modulation of differentiation. Recently it has been shown that ectopic expression of HH gene family members in vivo blocks chondrocyte maturation through activation of a parathyroid hormone related peptide (PTHrP) dependent negative regulatory loop in the perichondrium. However, the direct effect of HH on chondrocyte maturation has not been tested. Here, we studied the effect of retroviral overexpression of the chicken sonic hedgehog gene (Shh) on the growth and maturation of limb bud cells in micromass cultures. Shh is neither expressed nor required for the initiation of cellular condensation in normal micromass cultures. With Shh over-expression, micromass cultures developed novel tightly whorled nodules in addition to the normal Alcian Blue positive cartilage nodules. We characterized the new nodules and showed that they are strongly positive for alkaline phosphatase, enriched in type X collagen and weakly positive for Alcian Blue staining. Shh overexpression also increased cell proliferation, but this cannot account for the formation of the new nodules. This current study shows that misexpression of Shh in in vitro chondrogenic cultures promotes characteristics of hypertrophic chondrocytes. Thus HH has two complementary functions; a direct positive effect on chondrocyte hypertrophy in the absence of PTHrP pathway, and an indirect negative feedback loop through PTHrP to prevent other less differentiated chondrocytes from becoming hypertrophic. These two complementary actions of HH coordinate the progression of cartilage maturation.

Thyroid Follicular Hyperplasia Associated With Massive Extracellular Mucin Deposition

2017 ◽  
Vol 25 (6) ◽  
pp. 533-535 ◽  
Author(s):  
Francesco Nesa ◽  
Luca Poggi ◽  
Stefano Ferrero ◽  
Alessandro Del Gobbo
Keyword(s):  
Alcian Blue ◽  
Periodic Acid ◽  
The Other ◽  
Blue Staining ◽  
Alcian Blue Staining ◽  
Thyroid Tumors ◽  
Stromal Compartment ◽  
Follicular Hyperplasia ◽  
Nodular Hyperplasia ◽  
Thyroid Parenchyma

Extensive extracellular mucin deposition is a rare pathological thyroid condition with 6 cases described in literature so far. We report another case of a 67-year-old woman, discussing histopathological features, and review the literature. Our findings showed a diffuse mucin deposition in the stromal compartment of thyroid parenchyma. Histochemical stainings showed positivity for Alcian blue staining, but not for periodic acid–Shiff staining. Our case is peculiar because this mucin deposition was associated with benign nodular hyperplasia, in contrast with the other 6 reports, which described the same stromal alterations associated with benign or malignant thyroid tumors.

scholarly journals Alizarin Red S-Alcian Blue Staining for Regenerated tail of Common House Gecko (Hemidactylus frenatus)

2018 ◽  
Vol 7 (2) ◽  
pp. 57-59
Author(s):  
Rakhmiyati Rakhmiyati ◽  
Muhammad Ja’far Luthfi
Keyword(s):  
Alcian Blue ◽  
Histochemical Staining ◽  
Alizarin Red S ◽  
Blue Staining ◽  
Alcian Blue Staining ◽  
Tail Regeneration ◽  
Alizarin Red ◽  
Regenerate Tail ◽  
Hemidactylus Frenatus ◽  
Common House

Common House Gecko (Hemidactylus frenatus) is one of reptiles that have ability to autotomy their tails. Tail autotomy is a mechanism to protect it self from predators. After the tail broke, there will be wound healing on the tail which is then followed by a tail regeneration event. Original tail and regenerate tail is very different morphologically and anatomically. The original tail is composed of bones while the tail of the regenerate is composed of cartilage. Histochemical staining using Alizarin Red-S Alcian Blue was done to differentiate bone and cartilage. This method will stained bones red while the cartilage will stained blue.

The Pathobiology of Human Neuromas: An Electron Microscopic and Biochemical Study

1985 ◽  
Vol 10 (1) ◽  
pp. 49-53 ◽  
Author(s):  
MARIE A. BADALAMENTE ◽  
L. C. HURST ◽  
J. ELLSTEIN ◽  
C. A. McDEVITT
Keyword(s):  
Alcian Blue ◽  
Biochemical Study ◽  
Blue Staining ◽  
Alcian Blue Staining ◽  
Post Injury ◽  
Electron Microscopic ◽  
Biochemical Analyses

The formation of neuroma scar after trauma or neurorrhaphy is believed to be mediated by the response of collagen forming fibroblasts. In this study of twenty human neuromas, myofibroblasts were identified as a component of the scar. These cells occurred singly or as aggregates. There was a qualitative increase of myofibroblasts during the period from two to six months post-injury. From six months to one and one-half years post-injury, numbers and aggregations of myofibroblasts diminished, concurrent with collagen proliferation. Ultrastructural alcian blue staining and biochemical analyses revealed a glucosamine—glycosaminoglycan matrix within neuromas when compared to control nerves. Myofibroblasts appear to play a part in the pathobiology of human neuromas.

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