Interference of fixatives and fixation period on the morphologic analysis of ovarian preantral follicles

Zygote ◽  
2021 ◽  
pp. 1-4
Author(s):  
D.C.C. Brito ◽  
L.V.S. Ñaupas ◽  
S.S. Souza ◽  
G.L.H. Alcântara ◽  
J.R. Figueiredo ◽  
...  
Keyword(s):  
Stromal Cells ◽  
Histological Analysis ◽  
Fixation Time ◽  
Preantral Follicles ◽  
Carnoy’S Solution ◽  
Morphological Evaluation ◽  
Morphologic Analysis

Summary Ovine ovarian fragments (3 × 3 × 1 mm) were fixed in neutral buffered formalin (NBF), Carnoy’s solution (CAR), Davidson’s solution (DAV), or paraformaldehyde (PFA) for 12 h or 24 h. After this fixation time, each fragment was prepared for histological analysis. Although fixative and fixation period did not affect follicular and stromal cells density, the percentages of morphologically normal primordial and primary follicles was affected by the fixative type and period of fixation. Paraformaldehyde was not indicated as a fixative for ovarian fragments. Formalin was a suitable fixative only when the period of fixation was 12 h, while Carnoy was efficient after a fixation period of 24 h. In conclusion, the most indicated fixative for the morphological evaluation of ovarian preantral follicles was DAV, regardless of the fixation period, that is 12 or 24 h.

Follicle development in cryopreserved bitch ovarian tissue grafted to immunodeficient mouse

2012 ◽  
Vol 24 (3) ◽  
pp. 461 ◽  
Author(s):  
L. Commin ◽  
S. Buff ◽  
E. Rosset ◽  
C. Galet ◽  
A. Allard ◽  
...  
Keyword(s):  
Stromal Cells ◽  
Histological Analysis ◽  
Smooth Muscle Actin ◽  
Ovarian Tissue ◽  
Follicular Development ◽  
Slow Freezing ◽  
Angiogenic Factor ◽  
Muscle Actin ◽  
Once Daily

The present study evaluated: (1) in vivo follicular development in canine ovarian tissue after slow freezing and xenotransplantation; and (2) the use of erythropoietin (EPO) as an angiogenic factor to optimise the transplantation procedure. Frozen–thawed ovarian tissue from five bitches was grafted into severe combined immunodeficient (SCID) mice (n = 47) treated with or without EPO (500 IU kg–1, once daily for 3 days) (Groups A and B, respectively) and analysed after 0, 1, 8 or 16 weeks. Follicle grade, follicle density, follicle morphology and stromal cells density were assessed by histological analysis, whereas vascularisation of the graft was quantified by immunohistochemistry with anti-α-smooth muscle actin antibody. Despite a massive loss of follicles after grafting, secondary follicle density was higher at 8 and 16 weeks than at 1 week regardless of EPO treatment. EPO significantly improved early follicle morphology and stromal cell density after 8 weeks and blood vessel density at 16 weeks after transplantation (P < 0.05). Intact secondary follicles with more than three granulosa cells layers were observed 16 weeks after transplantation. The results suggest that canine ovarian tissue can be successfully preserved by our slow-freezing protocol because the tissue showed follicular growth after xenotransplantation. EPO treatment did not lessen the massive loss of follicles after transplantation.

Ectopic Osteogenesis of Bone Marrow Stromal Cells Loading in Hydroxyapatite/ β-Tricalcium Phosphate

2007 ◽  
Vol 330-332 ◽  
pp. 1109-1112 ◽  
Author(s):  
Yun Feng Lin ◽  
Ling Wu ◽  
Lei Liu ◽  
Ju Qiao ◽  
Wei Jing ◽  
...  
Keyword(s):  
Stromal Cells ◽  
Tricalcium Phosphate ◽  
Bone Marrow Stromal Cells ◽  
Histological Analysis ◽  
Marrow Stromal Cells ◽  
Cellular Phenotype ◽  
Rt Pcr ◽  
Sd Rats ◽  
Transmission Electron ◽  
Ectopic Bone

This study was to determine the ectopic osteogenic ability of BMSCs in combination with a scaffolding material comprising hydroxyapatite and β-tricalcium phosphate matrix (HA/β-TCP). BMSCs were obtained from the SD rats and induced to osteogenesis. Then these induced cells were seeded into HA/β-TCP and the constructs were auto-implanted subcutaneously for up to 12 weeks. Histological analysis, immunostaing, RT-PCR and transmission electron microscopy of the retrieved specimens at various intervals showed obvious trends of ectopic bone formation with obvious alteration of cellular phenotype.

252. Morphometric and histological analysis of ovaries from sheep heterozygous for the prolific woodlands allele

2005 ◽  
Vol 17 (9) ◽  
pp. 101
Author(s):  
E. S. Feary ◽  
J. L. Juengel ◽  
P. Smith ◽  
A. R. O'Connell ◽  
G. H. Davis ◽  
...  
Keyword(s):  
Histological Analysis ◽  
Follicular Development ◽  
Wild Type ◽  
Preantral Follicles ◽  
Antral Follicles ◽  
Mechanistic Pathway ◽  
Morphometric Methods ◽  
The Mean ◽  
Large Ovary

Woodlands are a line of Coopworth sheep with a novel, imprinted X-linked fecundity allele resulting in ovulation rates about 0.40 higher than wild-type animals. Daughters of progeny tested sires with and without the gene were studied. Previously, lambs heterozygous for the Woodlands allele were found to have larger ovaries and more antral (i.e. type 5) but not preantral (i.e. types 1–4) follicles than in wild-type contemporaries. The large ovary phenotype was found to be transient and was absent after puberty. However, based on follow-up studies it was evident that the large ovary phenotype was not strongly associated with the Woodlands fecundity allele. Thus, it was uncertain whether animals carrying the Woodlands gene had different follicular populations compared to wild-type controls. To address this question, follicular populations were compared in adult ewes heterozygous for the Woodlands allele with age-matched controls. Using standard morphometric methods and histological analysis, no differences were observed in the mean numbers of types 1, 1a, 2, 3 and 4 preantral follicles between the genotypes. Furthermore, no differences were observed between genotypes in follicular or oocyte diameters for any follicular type. The adult Woodlands carrier ewes had twice as many small type 5 follicles (< 1mm) when compared to wild-type contemporaries although no difference was seen in the numbers of antral follicles > 1mm in diameter. In addition, antrum formation occurred at a smaller follicular diameter in the heterozygous Woodlands animals. Therefore, the increased number of antral follicles observed in both lambs and adult ewes suggests that this difference in pattern of follicular development is associated with the X-linked fecundity allele. This novel phenotype of early antrum formation and larger number of small preantral follicles differs from that observed in sheep with the Inverdale or Booroola mutations, suggesting that a different mechanistic pathway is involved. Acknowledgements: The Marsden Fund, FRST and Ovita.

scholarly journals Fluorescent Nanodiamonds Enable Long-Term Detection of Human Adipose-Derived Stem/Stromal Cells in an In Vivo Chondrogenesis Model Using Decellularized Extracellular Matrices and Fibrin Glue Polymer

Polymers ◽  
2019 ◽  
Vol 11 (9) ◽  
pp. 1391 ◽  
Author(s):  
Yi-Chia Wu ◽  
Ya-Chin Wang ◽  
Wei-Ting Wang ◽  
Hui-Min David Wang ◽  
Hsin-Hung Lin ◽  
...  
Keyword(s):  
Fibrin Glue ◽  
Nude Mice ◽  
Stromal Cells ◽  
Histological Analysis ◽  
Control Group ◽  
Efficient Treatment ◽  
Fluorescent Nanodiamonds ◽  
Experimental Group

Clinically available materials, including allogeneic irradiated costal cartilage and fibrin glue polymer, were used as scaffolds for in vivo chondrogenic differentiation of human adipose-derived stem/stromal cells (hASCs) in the attempt to develop a more efficient treatment over current methods. Current studies include the use of growth-factor stimulation, tissue engineering, and biocompatible materials; however, most methods involve complicated processes and pose clinical limitations. In this report, the xenografts in the experimental group composed of a diced decellularized cartilage extracellular matrix (ECM), hASCs, and fibrin glue polymer were implanted into the subcutaneous layer of nude mice, and the results were compared with two groups of controls; one control group received implantation of decellularized cartilage ECM and fibrin glue polymer, and the other control group received implantation of hASCs mixed with fibrin glue polymer. To evaluate whether hASCs had in vivo chondrogenesis in the xenografts, hASCs were labeled with fluorescent nanodiamonds (FNDs), a biocompatible and photostable nanomaterial, to allow for long-term detection and histological analysis. Increased cellularity, glycosaminoglycan, and collagen deposition were found by the histological examination in the experimental group compared with control groups. With the background-free detection technique and time-gated fluorescence imaging, the numbers and locations of the FND-labeled hASCs could be detected by confocal microscopy. The chondrocyte-specific markers, such as aggrecan and type II collagen, were colocalized with cells containing signals of FNDs which indicated in vivo chondrogenesis of hASCs. Taken together, functional in vivo chondrogenesis of the hASCs could be achieved by clinically available decellularized cartilage ECM and fibrin glue polymer in the nude mice model without in vitro chondrogenic induction. The fluorescent signals of FNDs in hASCs can be detected in histological analysis, such as hematoxylin and eosin staining (H&E staining) without the interference of the autofluorescence. Our study may warrant future clinical applications of the combination of decellular cartilage ECM, fibrin glue polymer, and hASCs for cartilage repair.

scholarly journals Morphological evaluation duringin vitrochondrogenesis of dental pulp stromal cells

2012 ◽  
Vol 37 (1) ◽  
pp. 34 ◽  
Author(s):  
Choo-Ryung Chung ◽  
Ha-Na Kim ◽  
Yeul Park ◽  
Min-Jeong Kim ◽  
Young-Ju Oh ◽  
...  
Keyword(s):  
Stromal Cells ◽  
Dental Pulp ◽  
Morphological Evaluation

scholarly journals 7-T MRI tracking of mesenchymal stromal cells after lung injection in a rat model

2020 ◽  
Vol 4 (1) ◽  
Author(s):  
Stefania Rizzo ◽  
Francesco Padelli ◽  
Elena Rinaldi ◽  
Daniela Gioeni ◽  
Domenico Aquino ◽  
...  
Keyword(s):  
Mesenchymal Stromal Cells ◽  
Rat Model ◽  
Stromal Cells ◽  
Histological Analysis ◽  
Structured Illumination ◽  
Structured Illumination Microscopy ◽  
Target Organs ◽  
Magnetic Resonance Imaging Mri ◽  
Linear Decay

Abstract Background Mesenchymal stromal cells (MSCs) are able to migrate and engraft at sites of inflammation, injuries, and tumours, but little is known about their fate after local injection. The purpose of this study is to perform MSC tracking, combining in vivo 7-T magnetic resonance imaging (MRI) and histological assessment, following lung injection in a rat model. Methods Five lungs were injected with ferumoxide-labelled MSCs and five with perfluorocarbon-labelled MSCs and underwent 7-T MRI. MRI acquisitions were recorded immediately (T0), at 24 h (T24) and/or 48 h (T48) after injection. For each rat, labelled cells were assessed in the main organs by MRI. Target organs were harvested under sterile conditions from rats sacrificed 0, 24, or 48 h after injection and fixed for histological analysis via confocal and structured illumination microscopy. Results Ferumoxide-labelled MSCs were not detectable in the lungs, whereas they were not visible in the distant sites. Perfluorocarbon-labelled MSCs were seen in 5/5 injected lungs at T0, in 1/2 at T24, and in 1/3 at T48. The fluorine signal in the liver was seen in 3/5 at T0, in 1/2 at T24, and in 2/3 at T48. Post-mortem histology confirmed the presence of MSCs in the injected lung. Conclusions Ferumoxide-labelled cells were not seen at distant sites; a linear decay of injected perfluorocarbon-labelled MSCs was observed at T0, T24, and T48 in the lung. In more than half of the experiments, perfluorocarbon-labelled MSCs scattering to the liver was observed, with a similar decay over time as observed in the lung.

scholarly journals Effect of Ethylene Glycol on Structural Integrity at Each Stage of Preantral Follicle Development Post Vitrification of Rat Ovary-Histological Analysis

2021 ◽  
Vol 28 (4) ◽  
pp. 304-311
Author(s):  
Nova Anita ◽  
Abinawanto Abinawanto ◽  
Ahmad Aulia Jusuf ◽  
Anom Bowolaksono ◽  
Huriyah Adani Saoemi
Keyword(s):  
Ethylene Glycol ◽  
Structural Integrity ◽  
Histological Analysis ◽  
Follicle Development ◽  
Preantral Follicle ◽  
Preantral Follicles ◽  
Software Analysis ◽  
Ovarian Cortex ◽  
Rat Ovary ◽  
Grade 3

The structure of follicular tissue affects the ability to maintain the structural integrity of follicles against cryoinjury post-vitrification. Histological analysis was conducted on the structural integrity of each stage of preantral follicles post-vitrification using 7.5% and 15.0% doses of ethylene glycol (EG), and ovarian sections with HE staining were observed using an Olympus CX21 microscope connected to Optilab 3.0 lens and Image Raster software. Analysis was conducted on the ovarian cortex in the tracing line area using polygon measure tools to obtain follicle density (follicles/mm2) and follicle index (%) data. The result showed that the EG group 7.5% (KP1) increased follicle density compared to the vitrified group (KKV) in primordial (15.83±1.77) and primary (22.94±8.51) stages. Meanwhile, KP2 (EG 15%) was in primordial (41.92±6.45), primary (11.69±1.95), secondary (33.48±3.63), and tertiary (5.93±0.69) stages. KP1 increased grade 3 follicle index compared to KKV in primary (27.66±2.34), secondary (32.41±6.99), and tertiary (25.00±5.00) stages. Meanwhile, KP2 was in primary (26.87±6.68) and tertiary (25.00±5.00) stages. Both doses of 7.5% and 15.0% EG were able to maintain structural integrity at certain stages of preantral follicles. Secondary and tertiary follicles are the best stages in maintaining grade 3 follicular integrity with the addition of 7.5% EG.

Betaine-loaded CaCO3 microparticles improve survival of vitrified feline preantral follicles through higher mitochondrial activity and decreased reactive oxygen species

2020 ◽  
Vol 32 (5) ◽  
pp. 531
Author(s):  
D. C. C. Brito ◽  
S. F. S. Domingues ◽  
A. P. R. Rodrigues ◽  
L. M. Silva ◽  
K. A. Alves ◽  
...  
Keyword(s):  
Reactive Oxygen Species ◽  
Ascorbic Acid ◽  
Stromal Cells ◽  
Morphological Analysis ◽  
Ovarian Tissue ◽  
Reactive Oxygen ◽  
Mitochondrial Activity ◽  
Oxygen Species ◽  
Preantral Follicles ◽  
Sexually Mature

Ovary fragments from six sexually mature cats were vitrified in the presence or absence of betaine or ascorbic acid, loaded (7.4 or 74µM betaine; 20 or 200µM ascorbic acid) or not (1mM betaine or 0.3mM ascorbic acid) into CaCO3 microparticles, and assessed for follicular morphology, oxidative stress and mitochondrial activity Feline ovarian tissue was successfully preserved after vitrification in the presence of 74µM betaine loaded in CaCO3 microparticles, as confirmed by morphological analysis and the density of preantral follicles and stromal cells, as well as by the increased mitochondrial activity and decreased production of reactive oxygen species.

scholarly journals Medium and time conservation affect follicular morphology and superoxide dismutase enzyme activity of bovine preantral follicles during storage at 4ºC

2020 ◽  
Vol 41 (4) ◽  
pp. 1227
Author(s):  
Maiara Aline Gonçalves Ramos ◽  
Fabio Gallas Leivas ◽  
Daniele Missio ◽  
Francielli Weber Santos Cibin ◽  
Antônio Carlos Galarça Guimarães ◽  
...  
Keyword(s):  
Superoxide Dismutase ◽  
Saline Solution ◽  
The Other ◽  
Histological Evaluation ◽  
Control Group ◽  
Sod Activity ◽  
Preantral Follicles ◽  
Morphological Evaluation ◽  
Contrast Preservation

The aim of this study was to evaluate the morphology and superoxide dismutase enzyme (SOD) activity of bovine preantral follicles (PFs) preserved in TCM 199, saline solution or PBS at different conservation periods. Cow ovaries (n=6) were divided into 7 fragments. One small piece of each ovarian fragment was randomly removed to evaluate SOD activity, while the remainder was immediately fixed for morphological evaluation as a control group. The other 6 fragments were randomly distributed in tubes containing TCM 199, saline solution, or PBS and maintained at 4ºC for 6 or 24 h. For histological evaluation, the fragments were fixed in Carnoy and stained with PAS-hematoxylin, following being classified PFs in relation to their follicular morphology in normal or degenerated. Determination of SOD activity was based on the ability to inhibit autoxidation of adrenaline in adrenochrome. Evaluation of follicular morphology showed that follicles preserved in TCM 199 for 6 h did not differ from the control (P > 0.05). In contrast, preservation in saline solution and PBS for 6 or 24 h and TCM 199 for 24 h decreased normal PFs compared to the control (P < 0.05). SOD showed a lower activity in ovarian cortical tissue kept in TCM 199 for 6 h and saline solution for 24 h than in the other groups. Our study shows that incubation using TCM 199 at 4°C for 6 h can be used to efficiently conserve female bovine PFs in situ.

scholarly journals Structural and Ultrastructural Morphological Evaluation of Giant Anteater (Myrmecophaga Tridactyla) Prostate Gland

2021 ◽  
Author(s):  
Fernanda B C de Moura ◽  
Letícia H. T. S. Sampaio ◽  
Priscila E. Kobayashi ◽  
Renee Laufer-Amorim ◽  
João Carlos P. Ferreira ◽  
...  
Keyword(s):  
Stromal Cells ◽  
Periodic Acid ◽  
Prostate Gland ◽  
Ultrastructural Feature ◽  
Secretory Cells ◽  
Morphological Description ◽  
Basal Cells ◽  
Periodic Acid Schiff ◽  
Morphological Evaluation ◽  
Young Subjects

Abstract The giant anteater (Myrmecophaga tridactyla) is a vulnerable species from Central and South Americas, that is considered possibly extinct in Belize, Guatemala, El Salvador, and Uruguay. Due to the species conservations and reproduction’s importance, this research aimed to characterize the morphology, histochemical, immunohistochemical, and ultrastructural feature of the giant anteater prostate gland. For this, we collected 11 giant anteater prostate glands and performed macroscopic, morphological, histochemical, immunohistochemical, and ultrastructural analysis. Nine-prostate glands from adult and two from young subjects were studied. Grossly, the adult giant anteater prostate gland is divided in two distinct zones; the central zones composed mainly of ducts and the peripheral zones of acini formed by secretory cells. The secretory cells showed positive periodic acid–Schiff staining. Furthermore, the immunohistochemical characterization revealed a similar human prostate pattern, with p63 staining basal cells, UPIII superficial cells of prostatic urethra, AR expressing nucleus of secretory and stromal cells, and PSA staining prostatic epithelial cells. Overall, our research provided an indepth morphological description of the giant anteater prostate gland, providing a valuable information for futures studies focused on giant anteater conservation.

Sign in / Sign up

Export Citation Format

Share Document