scholarly journals Viability, yield and expansion capability of feline MSCs obtained from subcutaneous and reproductive organ adipose depots

2021 ◽  
Vol 17 (1) ◽  
Author(s):  
Amy Wysong ◽  
Priscilla Ortiz ◽  
Douglas Bittel ◽  
Lindsey Ott ◽  
Francis Karanu ◽  
...  
Keyword(s):  
Cell Viability ◽  
Subcutaneous Fat ◽  
Enzymatic Digestion ◽  
Reproductive Organs ◽  
Reproductive Organ ◽  
Negative Control ◽  
Allogeneic Mscs ◽  
Donor Tissue ◽  
Fat Depots ◽  
Expansion Capacity

Abstract Background The source of multipotent stromal cells (MSC) can have a significant influence on the health and expansion capacity of the cells. As the applications for allogeneic MSCs in the treatment of feline diseases increase, the location of the initial donor tissue must be analyzed. To date, comparisons have only been made between feline MSCs collected from bone marrow or abdominal fat. This is the first report to compare cells obtained from different adipose depots in the cat with a focus on clinically relevant donor tissues. The tissue was collected from 34 healthy cats undergoing spaying (fat around the ovaries and uterine horn) or subcutaneous fat collected during surgical procedures. Results The amount of starting material is essential to isolate sufficient MSCs. The total tissue yield from the subcutaneous fat was significantly greater than could be obtained from around the reproductive organs, leading to 3 times more MSCs per donor. However, the concentration of MSCs obtained from reproductive fat was higher than from subcutaneous fat. In addition, the viability of the MSCs from the reproductive fat was significantly higher than the subcutaneous fat. Since most spaying occurs in young cats (under 18 months) reproductive fat was collected from adult cats during spaying, illustrating that age did not alter the yield or viability of the MSCs. When sufficient tissue was collected, it was digested either mechanically or enzymatically. Mechanical digestion further decreased the viability and yield of MSCs from subcutaneous fat compared to enzymatic digestion. Biomarkers of stem cell characterization, expansion capacity and function were detected using qPCR. CD70, CD90 and CD105 were all expressed in high levels in the 3 groups. However, the reproductive fat had higher levels of CD73 with the mechanically digested subcutaneous fat having the least. Gata6 was detected in all samples while Sox2 and Sox17 were also detected with higher quantities found in the enzymatically digested subcutaneous fat. Negative control genes of Gata4 and Pdx1 showed no detection prior to 50 cycles. During the first three passages, age of the donor, location of the donor tissue, or digestion protocol had no effect on cell culture doubling times or cell viability. Conclusions While MSCs from reproductive fat had superior cells/tissue weight and initial viability, there were still dramatically fewer cells obtained compared to subcutaneous fat due to the limited amount of tissue surrounding the reproductive organs. Further, in P1-P3 cultures there were no differences noted in doubling time or cell viability between tissue obtained from reproductive or subcutaneous fat depots.

scholarly journals Viability, Yield and Expansion Capability of Feline MSCs Obtained from Subcutaneous and Reproductive Organ Adipose Depots

2020 ◽  
Author(s):  
Amy Wysong ◽  
Priscilla Ortiz ◽  
Douglas Bittel ◽  
Lindsey Ott ◽  
Francis Karanu ◽  
...  
Keyword(s):  
Adipose Tissue ◽  
Subcutaneous Fat ◽  
Enzymatic Digestion ◽  
Reproductive Organs ◽  
Reproductive Organ ◽  
Allogeneic Mscs ◽  
Significant Difference ◽  
And Function ◽  
Expansion Capacity ◽  
Negative Controls

Abstract Background: The source of multipotent stromal cells (MSC) can have a significant influence on the health and expansion capacity of the cells. As the applications for allogeneic MSCs in the treatment of feline diseases increase, the location of the initial donor tissue must be analyzed. To date, comparisons have only been made between feline MSCs collected from bone marrow or abdominal fat. This is the first report to compare cells obtained from different adipose depots in the cat. The adipose tissue was collected from 32 healthy cats undergoing spaying (fat around the ovaries and uterine horn) or subcutaneous fat collected during surgical procedures. Results: The total tissue yield from the subcutaneous fat was significantly greater than could be obtained from around the reproductive organs, leading to 3 times more total MSCs. However, the density of MSCs obtained from reproductive fat was significantly higher than from subcutaneous fat. In addition, the viability of the MSCs from the reproductive fat was significantly higher than the subcutaneous fat. When sufficient tissue was collected, it was digested either mechanically or enzymatically. Mechanical digestion further decreased the viability and yield of MSCs from subcutaneous fat compared to enzymatic digestion. Biomarkers of stem cell expansion and function were detected using qPCR. Gata6 was detected in all samples similarly regardless of site of harvest or method of digestion. Sox2 and Sox17 were also detected in all samples with higher quantities found in the enzymatically digested subcutaneous fat. Control genes of Gata4 and Pdx1 were detected in very low levels. Negative controls showed no detection prior to 50 cycles. During the first passage there was no statistically significant difference in doubling times or viability. But at P2, MSCs from the enzymatically-digested reproductive fat had the lowest doubling time and a trend towards the highest viability. Conclusion: Feline reproductive adipose tissue is a reasonable source of MSCs for eventual therapeutic applications with superior cell density, viability and expansion

Reproductive Organ Education for Psychic Health Development of Early Age

2018 ◽  
Vol 2 ◽  
Author(s):  
Dwi Darwati
Keyword(s):  
Early Childhood ◽  
Early Childhood Development ◽  
Deviant Behavior ◽  
Reproductive Organs ◽  
Reproductive Organ ◽  
The Body ◽  
Childhood Development ◽  
Early Age ◽  
Digital Era ◽  
Stage Of Development

Reproductive  health education should be given since early childhood by using language that is adapted to the stage of development. If you procrastinate and wait until the teenager it is already too late because in the days of the digital era, as now, all the information can be easily accessed by anyone including children early age. If the early childhood misinformed about their reproductive organs it would disrupt the physical and psychological development due to the wrong behavior in caring for and maintaining reproductive organs. Qur’an as the holy book of Muslims describes the steps of reproduction and  imparting education wisely as well as how to apply such education. This kind of education must be in accordance with the conditions of children and there should not be a lie about it We can also use media and methods such as pictures, songs, tap or other visual  media which can give clearer information, so that children can clearly see parts of the body, their characteristics, and how to treat and care them. The impropriate approach in conveying this kind of knowledge will be very dangerous for children. The provision of early age reproductive organs education can prevent the occurrence of deviant behavior as well as protect children from dangerous influence in early childhood development.

scholarly journals EFFECTS OF BREED AND SEX ON THE PATTERNS OF FAT DEPOSITION AND DISTRIBUTION IN SWINE

1980 ◽  
Vol 60 (2) ◽  
pp. 223-230 ◽  
Author(s):  
S. D. M. JONES ◽  
R. J. RICHMOND ◽  
M. A. PRICE ◽  
R. B. BERG
Keyword(s):  
Subcutaneous Fat ◽  
Fat Distribution ◽  
Allometric Equation ◽  
Muscle Weight ◽  
Constant Weight ◽  
Independent Variables ◽  
Fat Depots ◽  
Wide Range ◽  
Sex Type ◽  
Breed Differences

The growth and distribution of fat from 163 pig carcasses were compared among five breeds (Duroc × Yorkshire (D × Y), Hampshire × Yorkshire (H × Y), Yorkshire (Y × Y), Yorkshire × Lacombe-Yorkshire (Y × L-Y) and Lacombe × Yorkshire (L × Y)) and two sex-types (barrows and gilts) over a wide range in carcass weight. The growth pattern of fat and the fat depots were estimated from the allometric equation (Y = aXb) using side muscle weight and side fat weight separately as independent variables. Growth coefficients (b) for intermuscular and subcutaneous fat depots were similar for the hindquarter but the intermuscular depot coefficient was slightly higher for the forequarter. The coefficient for body cavity fat was highest in all comparisons. No significant differences were detected for coefficients among breeds and between sexes using both total muscle and total side fat as independent variables. Significant breed and sex-type differences were found in the fat depots at a constant weight of side muscle. This would indicate that breed differences in fatness seemed to be more influenced by the initiation of fattening at different muscle weights than by any inherent differences in rate of fattening. Significant breed differences were also found in the fat depots at a constant fat weight, indicating that breed may influence fat distribution. Sex-type had no effect on fat distribution when the evaluation was made at constant fatness.

scholarly journals Large increases in adipose triacylglycerol flux in Cushingoid CRH-Tg mice are explained by futile cycling

2013 ◽  
Vol 304 (3) ◽  
pp. E282-E293 ◽  
Author(s):  
Charles Harris ◽  
Donald J. Roohk ◽  
Mark Fitch ◽  
Benjamin M. Boudignon ◽  
Bernard P. Halloran ◽  
...  
Keyword(s):  
Protein Synthesis ◽  
Collagen Synthesis ◽  
Muscle Protein ◽  
Subcutaneous Fat ◽  
Skin Thickness ◽  
Breakdown Rate ◽  
Triglyceride Synthesis ◽  
Fat Depots ◽  
Breakdown Rates ◽  
Skin Collagen

Glucocorticoids are extremely effective anti-inflammatory therapies, but their clinical use is limited due to severe side effects, including osteoporosis, muscle wasting, fat redistribution, and skin thinning. Here we use heavy water labeling and mass spectrometry to measure fluxes through metabolic pathways impacted by glucocorticoids. We combine these methods with measurements of body composition in corticotropin-releasing hormone (CRH)-transgenic (Tg)+ mice that have chronically elevated, endogenously produced corticosterone and a phenotype that closely mimics Cushing's disease in humans. CRH-Tg+ mice had increased adipose mass, adipose triglyceride synthesis, and greatly increased triglyceride/fatty acid cycling in subcutaneous and abdominal fat depots and increased de novo lipogenesis in the abdominal depot. In bone, CRH-Tg+ mice had decreased bone mass, absolute collagen synthesis rates, and collagen breakdown rate. In skin, CRH-Tg+ mice had decreased skin thickness and absolute collagen synthesis rates but no decrease in the collagen breakdown rate. In muscle, CRH-Tg+ mice had decreased muscle mass and absolute protein synthesis but no decrease in the protein breakdown rate. We conclude that chronic exposure to endogenous glucocorticoid excess in mice is associated with ongoing decreases in bone collagen, skin collagen, and muscle protein synthesis without compensatory reduction (coupling) of breakdown rates in skin and muscle. Both of these actions contribute to reduced protein pool sizes. We also conclude that increased cycling between triglycerides and free fatty acids occurs in both abdominal and subcutaneous fat depots in CRH-Tg+ mice. CRH-Tg mice have both increased lipolysis and increased triglyceride synthesis in adipose tissue.

scholarly journals Evaluation of coconut water neutralized by different agents on the viability of human fibroblasts: an in vitro study

2016 ◽  
Vol 45 (4) ◽  
pp. 234-239 ◽  
Author(s):  
Priscilla Barbosa Ferreira SOARES ◽  
Aletheia Moraes ROCHA ◽  
Manuella Verdinelli de Paula REIS ◽  
Camilla Christian Gomes MOURA ◽  
Carlos José SOARES
Keyword(s):  
Cell Viability ◽  
Tap Water ◽  
Human Fibroblasts ◽  
In Vitro Study ◽  
Positive Control ◽  
Negative Control ◽  
Coconut Water ◽  
Ph Adjustment ◽  
Adjustment Method

Abstract Objective This study evaluated four types of pH adjustment of the coconut water (CW) on viability of human fibroblasts (HFF). Material and method Natural and industrialized CW were adjusted to pH 7.0 using: (1) Sodium Hidroxide (NaOH), (2) Sodium bicarbonate (NaHCO3), (3) Triethanolamine (C6H15NO3), (4) 2-Amino-2-Methil-1-Propanol (C4H11NO). Fibroblasts were plated at 2×104/ well in 96 well plates and maintained in the CW solutions for 2 h and 4 h. Positive control was represented by HFF maintained in DMEM and the negative control by tap water. Cell viability was analyzed by MTT formazan method. Data were analyzed by 3-way ANOVA followed by Tukey’s and Dunnet’s test. Result There are no significant effect on the cell viability regarding type of CW, period of evaluation, and the interactions between CW and period of evaluation, CW and pH adjustment method, pH adjustment method and period of evaluation (p>0.05). Conclusion The product used for CW pH adjustment did not influenced HFF viability, thought there are a tendency of better performance in natural CW.

scholarly journals Characterization of Human Knee and Chin Adipose-Derived Stromal Cells

2015 ◽  
Vol 2015 ◽  
pp. 1-11 ◽  
Author(s):  
Magali Kouidhi ◽  
Phi Villageois ◽  
Carine M. Mounier ◽  
Corinne Ménigot ◽  
Yves Rival ◽  
...  
Keyword(s):  
Stromal Cells ◽  
Expression Profiles ◽  
Donor Site ◽  
Subcutaneous Fat ◽  
Gene Expression Profiles ◽  
Fat Grafting ◽  
Human Knee ◽  
Fat Depots ◽  
Adipose Derived Stromal Cells ◽  
Molecular Matching

Animal study findings have revealed that individual fat depots are not functionally equivalent and have different embryonic origins depending on the anatomic location. Mouse bone regeneration studies have also shown that it is essential to match theHoxcode of transplanted cells and host tissues to achieve correct repair. However, subcutaneous fat depots from any donor site are often used in autologous fat grafting. Our study was thus carried out to determine the embryonic origins of human facial (chin) and limb (knee) fat depots and whether they had similar features and molecular matching patterns. Paired chin and knee fat depots were harvested from 11 subjects and gene expression profiles were determined by DNA microarray analyses. Adipose-derived stromal cells (ASCs) from both sites were isolated and analyzed for their capacity to proliferate, form clones, and differentiate. Chin and knee fat depots expressed a differentHOXcode and could have different embryonic origins. ASCs displayed a different phenotype, with chin-ASCs having the potential to differentiate into brown-like adipocytes, whereas knee-ASCs differentiated into white adipocytes. These results highlighted different features for these two fat sites and indicated that donor site selection might be an important factor to be considered when applying adipose tissue in cell-based therapies.

scholarly journals Association of fibroblast growth factor 21 with metabolic phenotype and fat depots in young adults with abdominal obesity

2020 ◽  
Vol 22 (12) ◽  
pp. 23-30
Author(s):  
Ekaterina A. Zheleznova ◽  
◽  
Juliya V. Zhernakova ◽  
Merab A. Shariia ◽  
Nataliia V. Blinova ◽  
...  
Keyword(s):  
Body Mass Index ◽  
Growth Factor ◽  
Waist Circumference ◽  
Young People ◽  
Fibroblast Growth Factor ◽  
Subcutaneous Fat ◽  
Metabolic Phenotype ◽  
Fibroblast Growth Factor 21 ◽  
Fibroblast Growth ◽  
Fat Depots

Fibroblast Growth Factor 21 (FGF21) is a hormone-like protein involved in the regulation of energy balance and glucose and lipid homeostasis. The study of the association of this factor with the metabolic phenotype – metabolically healthy (MHAO) and metabolically unhealthy abdominal obesity (AO) and different fat depots (visceral, subcutaneous, epicardial, perivascular) in young people is of undoubted scientific and practical interest. Aim. To determine serum FGF21 levels and match it with the distribution of adipose tissue in young people with AO. Outcomes and methods. The study enrolled 132 people (mean age 37.59±6.35 years). 3 groups were formed: 0th – 16 conditionally healthy volunteers; 1st –46 people of 40 years [34; 43] with MHAO; 2nd – 70 people of 40 years [35; 44] with metabolic syndrome (MS). All subjects underwent measurement of height, body weight, waist circumference, calculation of body mass index. The FGF21 levels (ELISA KIT, BCM Diagnostics, Germany), lipid profile, 2-hour glucose tolerance test, glucose, insulin, leptin, adiponectin levels and HOMA-IR were assessed. Daily monitoring of blood pressure was performed. The volumes of subcutaneous, visceral, perivascular, epicardial fat, as well as subcutaneous fat to visceral fat ratio were determined with computed tomography. Additionally, for subanalysis, all patients (132 people, mean age 37.59±6.35 years) were divided into 6 groups depending on the presence of AO and the number of risk factors (RF): AO-0/FR-0 (n=16); AO-1/FR-0 (n=3); AO-1/FR-1 (n=40); AO-1/FR-2 (n=37); AO-1/FR-3 (n=14); AO-1/FR-4 (n=5). In each group, FGF21 levels was assessed. Results. The FGF21 levels was significantly higher in the groups of persons with MHAO (294.4 pg/ml) and MS (245.7 pg/ml) compared with the control group (110.2 pg/ml); p=0.04 and p=0.05, respectively. According to the correlation analysis data, there was significant weak association of FGF21 with age (r=0.22, p≤0.05), waist circumference (r=0.18, p≤0.05), hip circumference (r=0.26, p≤0.05), body mass index (r=0.3, p≤0.01). FGF21 was found to be associated with vis-ceral (r=0.2, p≤0.05) and subcutaneous (r=0.2, p≤0.05) fat depots. A significant association of FGF21 with triglycerides (r=0.21, p≤0.05) and leptin (r=0.24, p≤0.05) was registered. The FGF21 level ≥345.8 pg/ml reflected a 3-fold increase in the risk of MS in young people (AuROC 0.74, sensitivity 78.6%, specificity 75.0%, p<0.0001). The FGF21 levels ≥294.4 pg/ml was a risk marker for MHAO (AuROC 0,70, sensitivity 67.4%, specificity 75.0%, p<0.0001). According to the results of subanalysis, a significant (p<0.01) increase in the FGF21 concentration was revealed in the groups with an increase in the number of MS components. Conclusions. The FGF21 levels increases with the worsening of the metabolic phenotype; its increase is seen long before the formation of MS (in persons with MHAO). FGF21 in young people is associated with visceral and subcutaneous fat depots, triglyceride levels and leptin. FGF21≥345.8 pg/ml can be considered a predictor of MS in young people, but further research is required.

scholarly journals The key role of a glucagon-like peptide-1 receptor agonist in body fat redistribution

2019 ◽  
Vol 240 (2) ◽  
pp. 271-286 ◽  
Author(s):  
Li Zhao ◽  
Chunfang Zhu ◽  
Meng Lu ◽  
Chi Chen ◽  
Xiaomin Nie ◽  
...  
Keyword(s):  
Lipid Metabolism ◽  
Body Fat ◽  
Visceral Fat ◽  
Subcutaneous Fat ◽  
Control Group ◽  
Acid Oxidation ◽  
High Dose ◽  
Fat Depots ◽  
Glucagon Like Peptide ◽  
Glucagon Like Peptide 1

Glucagon-like peptide-1 receptor agonists (GLP-1RAs) are an ideal therapy for type 2 diabetes and, as of recently, for obesity. In contrast to visceral fat, subcutaneous fat appears to be protective against metabolic diseases. Here, we aimed to explore whether liraglutide, a GLP-1RA, could redistribute body fat via regulating lipid metabolism in different fat depots. After being fed a high-fat diet for 8 weeks, 50 male Wistar and Goto-Kakizaki rats were randomly divided into a normal control group, a diabetic control group, low- and high-dose liraglutide-treated groups and a diet-control group. Different doses of liraglutide (400 μg/kg/day or 1200 μg/kg/day) or an equal volume of normal saline were administered to the rats subcutaneously once a day for 12 weeks. Body composition and body fat deposition were measured by dual-energy X-ray absorptiometry and MRI. Isotope tracers were infused to explore lipid metabolism in different fat depots. Quantitative real-time PCR and Western blot analyses were conducted to evaluate the expression of adipose-related genes. The results showed that liraglutide decreased visceral fat and relatively increased subcutaneous fat. Lipogenesis was reduced in visceral white adipose tissue (WAT) but was elevated in subcutaneous WAT. Lipolysis was also attenuated, and fatty acid oxidation was enhanced. The mRNA expression levels of adipose-related genes in different tissues displayed similar trends after liraglutide treatment. In addition, the expression of browning-related genes was upregulated in subcutaneous WAT. Taken together, the results suggested that liraglutide potentially redistributes body fat and promotes browning remodeling in subcutaneous WAT to improve metabolic disorders.

Carcass composition in four sheep breeds: the importance of type of breed and stage of maturity

1980 ◽  
Vol 30 (1) ◽  
pp. 135-152 ◽  
Author(s):  
J. D. Wood ◽  
H. J. H. MacFie ◽  
R. W. Pomeroy ◽  
D. J. Twinn
Keyword(s):  
Subcutaneous Fat ◽  
Carcass Composition ◽  
Carcass Weight ◽  
Relative Growth ◽  
Fat Depots ◽  
Intermuscular Fat ◽  
The Mean ◽  
Total Fat ◽  
Omental Fat ◽  
Mature Size

ABSTRACTIn order to investigate the effects of type of breed on carcass composition, an examination was made of 361 lambs from four breeds: Clun Forest and Colbred (termed ewe breeds); and Suffolk and Hampshire (termed ram breeds). The animals were in four carcass weight groups averaging 15, 17, 19 and 21 kg.Percentage subcutaneous fat was influenced more by carcass weight than by breed, whereas both carcass weight and breed had similar effects on percentage lean. At the mean carcass weight of 18 kg, Colbreds, the leanest breed, had a similar value for percentage lean (about 57 % of carcass tissue weight) to the carcasses over all breeds weighing 15 kg; and Cluns, the fattest breed, had a similar value (about 54%) to those weighing 21 kg. Since the ram breeds were intermediate in composition between the two ewe breeds there was no effect of type of breed on carcass composition. The breed differences were related to eventual mature size and to the stage of maturity at each carcass weight, as judged by body length and bone weight measurements. However, Colbreds were bigger and leaner than published estimates of their mature weight suggested. Humerus weight was a good predictor of lean or total fat weight, explaining 83 % ofvariation when used as a predictor along with carcass weight.Type of breed had a marked effect on internal fat deposition, the ewe breeds having heavier weights of both kidney knob and channel fat (KKCF) and caul fat (omental fat) than the ram breeds; and on the length oflimb bones, the ewe breeds having longer but thinner bones than the ram breeds. The order of the relative growth of the tissues and fat depots was: subcutaneous fat > caul fat > KKCF > intermuscular fat > lean > bone. Therefore, the internal fat depots were later maturing than intermuscular fat.The percentage of prime cuts in the carcass was not affected by carcass weight. Colbreds had significantly lower values than the other breeds. Suffolks had the lowest lean to bone ratio.

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