scholarly journals Morphological and functional characteristics of cell culture derived from the mouse nail unit

2017 ◽  
Vol 5 (1) ◽  
pp. 62-66
Author(s):  
O. Kalmukova ◽  
A. Ustymenko ◽  
T. Lutsenko ◽  
P. Klymenko ◽  
V. Kyryk
Keyword(s):  
Cell Culture ◽  
Growth Rate ◽  
Doubling Time ◽  
Cultured Cells ◽  
Primary Cultures ◽  
Population Doubling ◽  
Synthetic Activity ◽  
Population Doubling Time ◽  
Proliferative Potential

Nail unit is a complex anatomical structure that is capable of rapid growth and regeneration throughout the life. Such significant reparative potential is associated with the presence different types of stem and progenitor cells, whose biology remains one of the fundamental issues today. Taking into account the active search for new stem cell sources for cell therapy, the view of the nail unit as a potential site for the localization of undifferentiated cells with stem potency is topical problem.Purpose. The study was conducted with an objective to establish the morphological, morphometric and proliferative characteristics of cultured cells isolated from the mouse nail unit.Materials and methods. Primary cultures of cells were obtained from tissue sampling, which included areas of the proximal nail fold, nail matrix and onychodermis of the FVB mouse nail organ. Cells were cultured in DMEM:F12 medium with 15 % fetal bovine serum during 6 passages. We determined the colony-forming activity, the population growth rate and doubling time, measured the area of cells, nuclei, and calculated the nuclear-cytoplasmic ratio. For cell morphology analysis, we used staining with Bemer’s hematoxylin and eosin, Heidenhain’s iron hematoxylin and May-Grünwald stain.Results. According to the morphological analysis in vitro the cells from mouse nail unit are heterogeneous with high synthetic activity and a low nuclear-cytoplasmic ratio – the features characteristic of the low-differentiated cells. The population doubling time of the culture was 80 ± 6.5 hours on average, the fastest growing cells were at the 4th passage (63 ± 7 hours). The specific growth rate for cell culture is low (0.01 ± 0.0007).The colony forming efficiency at the 5th passage was only 4 %. A significant number of colonies was small with large poorly proliferative cells, which may indicate a production of large numbers of transitional progenitor cells.Conclusion. The obtained cell culture from the mouse nail unit according to the analysis of their morphology, morphometry and proliferative potential is heterogeneous and requires the further development of pure culture technologies for the detailed characterization of separate subpopulations of cells.

Paradoxically slow preadipocyte replication and differentiation in corpulent rats

1990 ◽  
Vol 258 (2) ◽  
pp. E368-E376 ◽  
Author(s):  
G. Shillabeer ◽  
J. M. Forden ◽  
J. C. Russell ◽  
D. C. Lau
Keyword(s):  
Adipose Tissue ◽  
Doubling Time ◽  
Tissue Growth ◽  
Population Doubling ◽  
Population Doubling Time ◽  
Sprague Dawley ◽  
Induced Differentiation ◽  
Sd Rats

We have investigated the in vitro rate of replication and differentiation of preadipocytes derived from lean (+/+) and obese (cp/cp) male JCR:LA-corpulent (cp) rats in an attempt to identify mechanisms that regulate adipose tissue growth. Cp/cp rats were twofold heavier than age-matched lean rats by 9-10 mo. Cp/cp-derived preadipocytes demonstrated an inherently slower rate of replication than +/+ preadipocytes (population doubling time: cp/cp 52.3 +/- 9.6 h vs. +/+ 19.7 +/- 1.6 h), although the preadipocyte pool in the cp/cp was significantly greater. Cp/cp preadipocytes were resistant to hormonally induced differentiation (19.9 +/- 9.4% of cells accumulated lipid) but differentiated when cocultured with mature adipocytes to the same extent as preadipocytes derived from Sprague-Dawley (SD) rats (cp/cp 48.4 +/- 15.2% vs. SD 52.2 +/- 11.9%). In contrast, SD preadipocytes did not differentiate in response to mature adipocytes from +/+ rats (13.8 +/- 5.2%). Our observations suggest that preadipocyte replication and maturation may not be controlled in a coordinated manner.

scholarly journals Development and population growth of Hydra viridissima Pallas, 1766 (Cnidaria, Hydrozoa) in the laboratory

2008 ◽  
Vol 68 (2) ◽  
pp. 379-383 ◽  
Author(s):  
FC. Massaro ◽  
O. Rocha
Keyword(s):  
Growth Rate ◽  
Population Growth ◽  
Generation Time ◽  
Doubling Time ◽  
Test Organism ◽  
Laboratory Culture ◽  
Individual Growth ◽  
Population Doubling ◽  
Population Doubling Time ◽  
The Individual

Hydras, the most representative freshwater Cnidaria, are of common occurrence in bodies of water in every continent except Antarctica. This study was planned with the aim of maintaining a population of Hydra viridissima in laboratory culture to enable the determination of the individual and population growth-rates of this species, as well as its population doubling time and generation time, with a view to employing these common animals as test-organisms in ecotoxicological assays. The organisms were maintained in reconstituted water at 20 ± 2 °C, illuminated at 800 lux with a photoperiod of 12 hours light: 12 hours dark, and were fed on neonates of the cladoceran Ceriodaphnia silvestrii (3 or 4 neonates per hydra, 3 times a week). The individual growth-rate (k) of the species was 0.43, the maximum length of the column 2.53 mm and the generation time 6.6 ± 1.5 days on average. The hydra population showed an intrinsic growth-rate (r) of 0.0468, according to the fitted curve, and a doubling time of 14.8 ± 2.63 days. Hydra viridissima is easy to grow in the laboratory and performs well in the conditions used in this study. It is thus a promising candidate test-organism for ecotoxicological studies.

Cell fate, morphogenetic movement and population kinetics of embryonic endoderm at the time of germ layer formation in the mouse

Development ◽  
1987 ◽  
Vol 101 (3) ◽  
pp. 627-652 ◽  
Author(s):  
K.A. Lawson ◽  
R.A. Pedersen
Keyword(s):  
Cell Fate ◽  
Doubling Time ◽  
Primitive Streak ◽  
Yolk Sac ◽  
Population Doubling ◽  
Population Doubling Time ◽  
Germ Layer ◽  
Posterior Half ◽  
Visceral Yolk Sac

The fate of the embryonic endoderm (generally called visceral embryonic endoderm) of prestreak and early primitive streak stages of the mouse embryo was studied in vitro by microinjecting horseradish peroxidase into single axial endoderm cells of 6.7-day-old embryos and tracing the labelled descendants either through gastrulation (1 day of culture) or to early somite stages (2 days of culture). Descendants of endoderm cells from the anterior half of the axis were found at the extreme cranial end of the embryo after 1 day and in the visceral yolk sac endoderm after 2 days, i.e. they were displaced anteriorly and anterolaterally. Descendants of cells originating over and near the anterior end of the early primitive streak, i.e. posterior to the distal tip of the egg cylinder, were found after 1 day over the entire embryonic axis and after 2 days in the embryonic endoderm at the anterior intestinal portal, in the foregut, along the trunk and postnodally, as well as anteriorly and posteriorly in the visceral yolk sac. Endoderm covering the posterior half of the early primitive streak contributed to postnodal endoderm after 1 day (at the late streak stage) and mainly to posterior visceral yolk sac endoderm after 2 days. Clonal descendants of axial endoderm were located after 2 days either over the embryo or in the yolk sac; the few exceptions spanned the caudal end of the embryo and the posterior yolk sac. The clonal analysis also showed that the endoderm layer along the posterior half of the axis of prestreak- and early-streak-stage embryos is heterogeneous in its germ layer fate. Whereas the germ layer location of descendants from anterior sites did not differ after 1 day from that expected from the initial controls (approx. 90% exclusively in endoderm), only 62% of the successfully injected posterior sites resulted in labelled cells exclusively in endoderm; the remainder contributed partially or entirely to ectoderm and mesoderm. This loss from the endoderm layer was compensated by posterior-derived cells that remained in endoderm having more surviving descendants (8.4 h population doubling time) than did anterior-derived cells (10.5 h population doubling time). There was no indication of cell death at the prestreak and early streak stages; at least 93% of the cells were proliferating and more than half of the total axial population were in, or had completed, a third cell cycle after 22 h culture.(ABSTRACT TRUNCATED AT 400 WORDS)

scholarly journals Molecular Aspects of Adipose-Derived Stromal Cell Senescence in a Long-Term Culture: A Potential Role of Inflammatory Pathways

2020 ◽  
Vol 29 ◽  
pp. 096368972091734 ◽  
Author(s):  
Marta Pokrywczynska ◽  
Małgorzata Maj ◽  
Tomasz Kloskowski ◽  
Monika Buhl ◽  
Daria Balcerczyk ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Stem Cells ◽  
Doubling Time ◽  
Population Doubling ◽  
Population Doubling Time ◽  
Proliferative Potential ◽  
Differentiation Potential ◽  
Long Term Culture ◽  
Stromal Stem Cells

Long-term culture of mesenchymal stromal/stem cells in vitro leads to their senescence. It is very important to define the maximal passage to which the mesenchymal stromal/stem cells maintain their regenerative properties and can be used for cellular therapies and construction of neo-organs for clinical application. Adipose-derived stromal/stem cells were isolated from porcine adipose tissue. Immunophenotype, population doubling time, viability using bromodeoxyuridine assay, MTT assay, clonogencity, β-galactosidase activity, specific senescence-associated gene expression, apoptosis, and cell cycle of adipose-derived mesenchymal stromal/stem cells (AD-MSCs) were analyzed. All analyses were performed through 12 passages (P). Decreasing viability and proliferative potential of AD-MSCs with subsequent passages together with prolonged population doubling time were observed. Expression of β-galactosidase gradually increased after P6. Differentiation potential of AD-MSCs into adipogenic, chondrogenic, and osteogenic lineages decreased at the end of culture (P10). No changes in the cell cycle, the number of apoptotic cells and expression of specific AD-MSC markers during the long-term culture were revealed. Molecular analysis showed increased expression of genes involved in activation of inflammatory response. AD-MSCs can be cultured for in vivo applications without loss of their properties up to P6.

scholarly journals Cryopreservation of Iranian Markhoz Goat Fibroblast Cells as an Endangered National Genetic Resource

2021 ◽  
Author(s):  
Zahra Elyasi Gorji ◽  
Parvaneh Farzaneh ◽  
Ahmad Nasimian ◽  
Meysam Ganjibakhsh ◽  
Mehrnaz Izadpanah ◽  
...  
Keyword(s):  
Cell Lines ◽  
Genetic Resource ◽  
Cultured Cells ◽  
Local Population ◽  
Capra Hircus ◽  
Population Doubling ◽  
Population Doubling Time ◽  
Fibroblast Cells ◽  
Primary Fibroblast

Abstract The sustainable use of local animals is being eroded annually. Thus, a strategic vision for the conservation of biodiversity is of far-reaching emphasis to deal with unprecedented challenges in the local population growth process facing in the future. This study aimed to establish, characterize and cryopreserve endangered Markhoz goat (Capra hircus) fibroblast cell lines in vitro. These primary fibroblast cells were isolated from 58 Iranian Markhoz goats and individually cultured by explant technique in DMEM media supplemented with 10 % FBS. The cultured cell lines were morphologically consistent with fibroblast cells. The population doubling time for DMEM-cultured cells were 23±0.5 h. Chromosomal analysis indicated a total chromosome number of 2n = 60 with >95% frequency. Experimental assays for bacteria, fungi, yeast, and mycoplasma were reported negative. The efficiencies of VSV-G (pMDG) and lentiviral pCSGW vectors encoding fluorescent proteins showed an approximate value of 65%. Species identification for each sample was performed and confirmed correct goat cell banking without any miss- and cross-contamination. This study demonstrated the successful establishment of genetically stable fibroblast bank as a valuable genetic resource for endangered Iranian Markhoz goat breed.

scholarly journals AKTIVITAS EKSTRAK TEMULAWAK (Curcuma xanthorrhiza Roxb.) TERHADAP PROLIFERASI DAN DIFERENSIASI SEL OTAK BESAR ANAK TIKUS BERUMUR TIGA HARI SECARA IN VITRO

2011 ◽  
Vol 1 (2) ◽  
pp. 1-9
Author(s):  
Min Rahminiwati ◽  
Ita Juwita ◽  
Ani Murtisari ◽  
Latifah K Darusman
Keyword(s):  
Doubling Time ◽  
Population Doubling ◽  
Population Doubling Time ◽  
Curcuma Xanthorrhiza

Kurkumin yang terdapat dalam rimpang temulawak, selain dapat menginduksi terjadinya proliferasi sel progenitor pada otak tikus dewasa juga dapat menghambat kerja enzim tirosinkinaseyang berperan penting dalam mengatur pertumbuhan dan diferensiasi sel. Meskipun demikianrespon sel saraf terhadap ekstrak temulawak pada masa pertumbuhan perlu kajian lebih lanjut. Efekekstrak temulawak terhadap proliferasi dan diferensiasi sel otak besar atau serebrum pada masapertumbuhan anak diteliti pada sel otak anak tikus Spague Dawley berumur tiga hari yangditumbuhkan dalam media DMEM (Dulbeccos Modified Eagles Medium). Perlakuan dikelompokkan dalam kelompok kontrol positif (mDMEM+30 g/mL asiaticoside (AC), kontrolnegatif (mDMEM), kelompok yang memperoleh ekstrak temulawak (CZ) 100 ppm (mDMEM+100ppm CZ), CZ 200 ppm (mDMEM+200 ppm CZ), dan CZ 400 ppm (mDMEM+400 ppm CZ). Kultur diinkubasi pada suhu 37 o C dalam inkubator CO 5 % selama 6 hari. Parameter yang diamatiadalah population doubling time, komposisi sel saraf dan sel glia, panjang akson dan dendrite yangdiukur masing masing menggunakan hemositometer, pewarnaan Hematoxyilin Eosin (HE) danmikrometer. Hasil penelitian menunjukkan ekstrak temulawak pada konsentrasi 100 ppmmemperlambat prolfperasi, pada konsentrasi 400 ppm meningkatkan diferensiasi sel yangditunjukkan dengan meningkatnya ratio sel glia terhadap sel saraf dan mempengaruhi panjangakson dan dendrite.Kata kunci : Curcuma xanthorrhiza Roxb., neuron, sel glia, dendrite

Research on Biological Characteristics of Chronic Myelogenous Leukemia Patients’ Bone Marrow Mesenchymal Stem Cells.

Blood ◽  
2007 ◽  
Vol 110 (11) ◽  
pp. 4535-4535
Author(s):  
Xiao Chen Chen ◽  
De Pei Wu ◽  
Feng Chen ◽  
Wei Rong Chang
Keyword(s):  
Bone Marrow ◽  
Doubling Time ◽  
Tumor Volume ◽  
K562 Cells ◽  
Myelogenous Leukemia ◽  
Population Doubling ◽  
Population Doubling Time ◽  
Proliferation And Apoptosis ◽  
Marrow Mesenchymal Stem Cells

Abstract Objective To isolate and culture bone marrow mesenchymal stem cells (MSC) from chronic myelogenous leukemia (CML) patients and examine their functional characteristics; To investigate their biological characteristics like proliferation, apoptosis and tumorigenicity;To observe their effects on proliferation and apoptosis of K562 cells. Methods Bone marrow was extracted from the anterior superior iliac spines of 20 patients with CML. MSC were isolated and cultured in vitro and subcultured for three to five generations. Cell morphology was observed under microscope. The ultrastructure was observed with electron microscope. The immunophenotype was detected by flow cytometry (FCM). Different agents were used to induce the MSC to differentiate into osteocyte and adipocyte, and Von Kossa staining, oil-red staining was used to examine the ability of differentiation. The growth curve, cell cycle and apoptosis were investigated. MSC were cultured in soft agar for 2 weeks to observe the clone growth. BALB/C nude mice were inoculated with MSC to observe the tumorigenicity. MSC and K562 cells were cocultured in vitro and coinjected into subcutaneous of BALB/C nude mice, to observe the effects of MSC on proliferation and apoptosis of K562 cells. Results Fibroblast-like, positive in CD44, CD73 and CD90,and negative in CD34, CD45 and HLA-DR, the CML derived MSC could differentiate into osteocyte and adipocyte. The MSC showed normal ultrastructure. After 2 weeks’ culture, no clone was formed from the MSC. Four weeks after, no tumor was seen in the mice inoculated with MSC. The population doubling time of P3 MSC is (32.61±1.54)h. The population doubling time of P4 MSC is (32.59±1.23)h, The population doubling time of P5 MSC is (32.41±0.75)h. To investigate the cell cycle of the third passage MSC by FCM, there were(93.67%±1.66%)cells in phase G0/G1,(6.33%±1.66%)in phase S+G2+M. There weren’t any apoptosis observed. K562 cells adhensively cultivated with MSC were accelerated and the population doubling time decreased([29.59±0.46]h vs [37.49±2.19]h, P<0.05),cells in G0/G1 decreased ([16.43%±1.67%] vs [32.23%±3.35%], P<0.01),cells in phase S increased ([69.63%±3.09%] vs [59.37%±4.40%], P<0.05),and those in G2/M increased([13.93%±1.45%] vs [8.40%±1.05%], P<0.01),compared with that cultivated in suspension. The apoptosis in both conditions were not observed. Coinjected with MSC, K562 cells developed tumors in 100% of the mice by (12.00±0.82)d compared to (15.50±0.58)d when implanted alone, indicating a statistically earlier onset of tumor growth in presence of MSC (P<0.01). The tumor volume and tumor weight were also increased when K562 cells coinjected with MSC compared with implanted alone (K562+MSC vs K562):tumor volume([75.70±7.30]mm2 vs [37.38±2.39]mm2, P<0.01),tumor weight([0.64±0.08]g vs [0.32±0.06]g, P<0.01). Conclusion MSC from the bone marrow of CML patients can be isolated and cultured;MSC derived from CML seem to have the abilities of powerful proliferation in vitro and multiple differentiation and have no tumorigenicity. The MSC can accelerate the proliferation of K562 cells both in vitro and vivo.

Growth kinetics and phenotypic markers expression in human dermal stem cells

2021 ◽  
Vol 16 (12) ◽  
pp. 24-29
Author(s):  
Vinutha Eshwara Swamy ◽  
Nikhil Shetty ◽  
Jayaprakasha Shetty ◽  
Veena Shetty ◽  
Tonita Noronha ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Stem Cells ◽  
Growth Kinetics ◽  
Primary Cultures ◽  
Autologous Transplantation ◽  
Explant Culture ◽  
Population Doubling ◽  
Population Doubling Time ◽  
Proliferative Potential ◽  
Phenotypic Markers

Human dermal stem cells (DSCs) have generated significant interest in the field of regenerative medicine due to their prospects of autologous transplantation. The present study evaluated the growth kinetics and phenotypic markers expression in human DSCs. The primary cultures of DSCs (n=3) were established by explant culture and characterization of the cells was carried out by assessing morphology, viability, proliferation rate, population doubling time (PDT), cell cycle status and the expression of cell surface markers such as CD29, CD73, CD90 and CD166. The cells released from tissue explants showed spindleshaped fibroblast morphology with the mean percentage viability varying between 93.43% and 100% from passages 1 to 4. DSCs displayed a strong and steady proliferative potential with an average PDT of 42.55 hrs. Cell cycle profile of DSCs demonstrated the majority of cells (59.80% to 76.29%) at G0/G1 phase. Further, the phenotypic profile of markers confirmed the stromal origin of DSCs by exhibiting positivity for CD29, CD73, CD90 and CD166. In conclusion, the growth kinetics and expression of phenotypic markers are consistent with the notion that skin dermis contains a population of stem cells and can serve as a potential autologous source for therapeutic applications.

Creating our Future — Some Concerns of an Environmentalist

1994 ◽  
Vol 1 (1) ◽  
pp. 83-86
Author(s):  
George Lewis
Keyword(s):  
Growth Rate ◽  
Population Growth ◽  
Rapid Growth ◽  
Human Population ◽  
Doubling Time ◽  
Gold Coast ◽  
Population Doubling ◽  
Western World ◽  
Population Doubling Time ◽  
The Media

Australia is often described as having the fastest human population growth in the western world at 1.4 per cent per annum (ABS 1993). Queensland's growth rate in comparison is 2.52 per cent while the Gold Coast is currently experiencing 6.1 per cent which is equivalent to a population doubling time of 11 years. While such rapid growth is lauded in the media and discussed in glowing terms by economists, engineers and entrepreneurs, there are many others who in recent years have queried the wisdom of such rapidly increasing growth. In 1992 a group calling themselves The Union of Concerned Scientists, ‘comprising 1575 scientists from 69 countries, including a majority of Nobel laureates’ (Caswell 1) issued a ‘Warning to Humanity’ regarding the accelerating damage threatening humanity's global support systems. According to this group, ‘no more than one or a few decades remain before the chance to avert the threats we now confront will be lost and the prospects for humanity immeasurably diminished’. They refer specifically to over-consumption, poverty and spiralling populations: all of which we have in south-east Queensland.

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