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Differential calcium requirements for growth of mouse skin epithelial and fibroblast cells. 小鼠皮肤上皮细胞和成纤维细胞生长对钙的不同需求。
Pub Date : 1984-09-01 DOI: 10.1111/j.1365-2184.1984.tb00611.x
M F Kulesz-Martin, D Fabian, J S Bertram

Concentration of extracellular Ca++ optimum for growth of cell types of mesodermal origin have been reported to be up to 100-fold higher than concentrations optimal for epidermal or other epithelial lining cells. In order to examine Ca++ requirements of epithelial v. fibroblastic cells derived from a common tissue source, prior to prolonged culture, freshly isolated mouse epidermal keratinocytes, hair follicle cells and dermal fibroblasts were plated at high density or at clonal density in medium ranging from 0.014 to 1.4 mM Ca++. Epithelial skin cells grew best at Ca++ levels below 0.1 mM while dermal fibroblasts grew best at a Ca++ concentration of 1.4 mM. The epithelial cell types exhibited marked morphologic changes in response to Ca++, while the fibroblasts did not. These results suggest that the variations in Ca++ response between lining epithelium and mesenchymal cells resulted from inherent differences in these cell types, but a mechanism for such differential effects has not yet been defined.

据报道,中胚层来源的细胞类型生长的最佳细胞外钙离子浓度比表皮细胞或其他上皮细胞的最佳浓度高100倍。为了检验来自同一组织来源的上皮细胞和成纤维细胞对Ca++的需求,在长时间培养之前,将新鲜分离的小鼠表皮角质形成细胞、毛囊细胞和真皮成纤维细胞以高密度或克隆密度镀在0.014至1.4 mM Ca++的培养基中。在Ca++浓度低于0.1 mM时,皮肤上皮细胞生长最好,而在Ca++浓度为1.4 mM时,真皮成纤维细胞生长最好。上皮细胞类型对Ca++的反应表现出明显的形态变化,而成纤维细胞则没有。这些结果表明,衬里上皮细胞和间充质细胞之间ca2 ++反应的差异是由这些细胞类型的内在差异引起的,但这种差异效应的机制尚未明确。
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引用次数: 28
Liver cell hyperplasia: on the suitability of using the metaphase-arrest technique. 肝细胞增生:中期阻滞技术应用的适用性。
Pub Date : 1984-09-01 DOI: 10.1111/j.1365-2184.1984.tb00610.x
J A Alabi, M R Alison

This study has explored the possibility of applying the metaphase-arrest method with colchicine to two models of induced liver growth in the rat, regenerative growth and phenobarbital-induced growth. At a dosage of 0.5 mg/kg body weight (BW), colchicine caused a linear accumulation of mitoses for up to 90 min when administered at 3 days after the start of phenobarbital treatment; however these mitoses included a number of anaphases and telophases. No anaphase escape was seen when this dose of colchicine was given at various times after partial hepatectomy, though the arrested mitoses were invariably more fragmented and some may have even degenerated beyond recognition as early as 90 min after injection. It is concluded that the optimal dose of stathmokinetic agent is heavily dependent on the relative liver weight, and thus would change continuously during compensatory hyperplasia.

本研究探讨了秋水仙碱中期阻滞法应用于大鼠肝脏再生生长和苯巴比妥诱导生长两种模型的可能性。当剂量为0.5 mg/kg体重(BW)时,在苯巴比妥治疗开始后3天给药时,秋水仙碱引起有丝分裂的线性积累长达90分钟;然而,这些有丝分裂包括许多后期和末期。在部分肝切除术后的不同时间给予这种剂量的秋水仙碱,没有观察到后期逃逸,尽管停止的有丝分裂总是更加碎片化,有些甚至可能早在注射后90分钟就退化到无法辨认。由此可见,稳定剂的最佳剂量与肝脏相对重量有很大关系,因此在代偿性增生过程中稳定剂的最佳剂量会持续变化。
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引用次数: 0
Aspects of statistics in studies of cell proliferation. I. Multistage sampling. 细胞增殖研究中的统计学方面。1、多级采样。
Pub Date : 1984-09-01 DOI: 10.1111/j.1365-2184.1984.tb00613.x
D R Appleton

In cell kinetic experiments, as in many other branches of biological science, we often sample at various stages of an experiment: we may for example take a sample of animals, then from each study a sample of sites, and from each site take replicate observations. This sampling process can be optimized to give maximum precision to an estimated quantity, but care must be taken in analysing data so gathered because the analysis depends on the precise sampling strategy.

在细胞动力学实验中,正如在生物科学的许多其他分支中一样,我们经常在实验的不同阶段取样:例如,我们可以取动物样本,然后从每次研究中取一个地点样本,并从每个地点进行重复观察。这种采样过程可以优化,使估计的数量具有最大的精度,但在分析这样收集的数据时必须小心,因为分析依赖于精确的采样策略。
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引用次数: 2
The metaphase-arrest technique applied to human cervical epithelium. I. Technique and dose-response studies. 中期阻滞技术在人宫颈上皮中的应用。1 .技术和剂量反应研究。
Pub Date : 1984-09-01 DOI: 10.1111/j.1365-2184.1984.tb00609.x
D Ireland, J M Monaghan

In order to study cell proliferation in intact human cervical epithelium, a technique involving metaphase arrest has been utilized. Metaphase accumulation is observed following intraepithelial administration of vincristine sulphate, at a predetermined optimum dose of 50 micrograms/ml. A significant delay before the onset of stathmokinetic activity is demonstrated; thereafter linearity of accumulation is apparent over a 4-hr period following injection. The technique appears applicable to the estimation and comparison of cell production rates in vivo of normal cervical epithelium, cervical intraepithelial neoplasia, wart-affected cervical epithelium and early invasive carcinoma. However, individual values are likely to be imprecise in isolation, indicating the need to study relatively large numbers of subjects in each group. Some practical difficulties are discussed.

为了研究完整人宫颈上皮细胞的增殖,采用了一种涉及中期阻滞的技术。在预先确定的最佳剂量为50微克/毫升的硫酸长春新碱上皮内给药后,观察到中期积累。在静动力活动开始前有明显的延迟;此后,在注射后的4小时内,积累的线性是明显的。该技术似乎适用于正常宫颈上皮、宫颈上皮内瘤变、疣感染宫颈上皮和早期浸润性癌的体内细胞生成率的估计和比较。然而,单独的个体值可能是不精确的,这表明需要在每组中研究相对大量的受试者。讨论了一些实际困难。
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引用次数: 1
Mirex-induced adaptive liver growth: a corticosterone-mediated response. mirex诱导的适应性肝脏生长:皮质酮介导的反应。
Pub Date : 1984-09-01 DOI: 10.1111/j.1365-2184.1984.tb00605.x
J D Yarbrough, L D Brown, J M Grimley

The adaptive liver growth response was investigated in intact and adrenalectomized rats. When adult male rats were given a single oral dose of mirex (100 mg/kg body weight) there was a 72% increase in relative liver weight (RLW) in 72 hr. Based on [3H]-thymidine [( 3H]TdR) incorporation into hepatic DNA, there was also a wave of DNA synthesis which peaked at 48 hr and decreased to essentially control values by 96 hr post mirex dose. In mirex-dosed adrenalectomised (Adx) animals, the RLW was increased by only 38% and there was sustained DNA synthesis. When mirex-dosed Adx rats were given corticosterone supplements, the RLW response was similar to the RLW response in intact mirex-dosed rats. However, the 48-hr DNA synthesis peak seen in intact mirex-dosed rats was eliminated. From these data it is suggested that mirex-induced adaptive liver growth has two components: a hypertrophic component which is mediated by corticosterone, and a hyperplastic component which is independent of corticosterone.

研究了正常大鼠和去肾上腺大鼠肝脏的适应性生长反应。成年雄性大鼠单次口服米雷克斯(100 mg/kg体重),72小时内相对肝脏重量(RLW)增加72%。基于[3H]-胸腺嘧啶[(3H) TdR)掺入肝脏DNA,也有一波DNA合成,在48小时达到峰值,在给药96小时后下降到基本控制值。在混合剂量肾上腺切除(Adx)动物中,RLW仅增加38%,并且DNA合成持续。当混合剂量的Adx大鼠给予皮质酮补充剂时,RLW反应与完整混合剂量大鼠的RLW反应相似。然而,在完整的mirex剂量大鼠中看到的48小时DNA合成峰被消除。从这些数据表明,混合液诱导的适应性肝脏生长有两个组成部分:由皮质酮介导的肥厚成分和独立于皮质酮的增生性成分。
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引用次数: 14
Progressive increase in polyamine levels in 9L cells in vitro during the cell cycle: comparison between cells isolated by centrifugal elutriation and cells grown in synchrony. 体外9L细胞在细胞周期中多胺水平的逐渐增加:离心洗脱分离的细胞与同步生长的细胞的比较。
Pub Date : 1984-09-01 DOI: 10.1111/j.1365-2184.1984.tb00602.x
S M Oredsson, J W Gray, L J Marton

Centrifugal elutriation was used to separate 9L rat brain tumour cells into fractions enriched in the G1, S, or G2/M phases of the cell cycle. Cells enriched in early G1 phase were recultured, grown in synchrony, and harvested periodically for analysis of their DNA distribution and polyamine content. Mathematical analysis of the DNA distributions indicated that excellent synchrony was obtained with low dispersion throughout the cell cycle. Polyamine accumulation began at the time of seeding, and intracellular levels of putrescine, spermidine, and spermine increased continuously during the cell cycle. In cells in the G2/M phase of the cell cycle, putrescine and spermidine levels were twice as high as in cells in the G1 phase. DNA distribution and polyamine levels were also analysed in cells taken directly from the various elutriation fractions enriched in G1, S, or G2/M. Because we did not obtain pure S or G2/M populations by elutriation or by harvesting synchronized cells, a mathematical procedure--which assumed that the measured polyamine levels for any population were linearly related to the fraction of cells in the G1, S, and G2/M phases times the polyamine levels in these phases and that polyamine levels did not vary within these phases--was used to estimate 'true' phase-specific polyamine levels (levels to be expected if perfect synchrony were achieved). Estimated 'true' phase-specific polyamine levels calculated from the data obtained from cells either sorted by elutriation or obtained from synchronously growing cultures were very similar.

采用离心洗脱将9L大鼠脑肿瘤细胞分离成细胞周期G1、S或G2/M期富集的部分。重新培养G1期早期富集的细胞,同步生长,并定期收获以分析其DNA分布和多胺含量。DNA分布的数学分析表明,在整个细胞周期中获得了良好的同步性和低分散。多胺积累始于播种时期,细胞内腐胺、亚精胺和精胺水平在细胞周期中不断增加。在细胞周期G2/M期的细胞中,腐胺和亚精胺的水平是G1期细胞的两倍。还分析了直接从G1、S或G2/M富集的不同洗脱组分中提取的细胞中的DNA分布和多胺水平。由于我们没有通过洗脱或收获同步细胞获得纯S或G2/M群体,因此使用数学程序(假设任何群体的测量多胺水平与G1, S和G2/M阶段的细胞比例乘以这些阶段的多胺水平线性相关,并且多胺水平在这些阶段内没有变化)来估计“真正的”阶段特异性多胺水平(如果达到完美同步则预期的水平)。从通过洗脱分选的细胞或从同步培养获得的细胞中获得的数据计算出的“真实”相特异性多胺水平非常相似。
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引用次数: 9
Changes in the DNA labelling index after beta irradiation of the mouse epidermis. 小鼠表皮辐照后DNA标记指数的变化。
Pub Date : 1984-09-01 DOI: 10.1111/j.1365-2184.1984.tb00606.x
L S Hansen, J E Coggle, J Wells, M W Charles

This study looked at the changes in the interfollicular DNA labelling index (LI) with time after strontium-90/yttrium-90 beta irradiation of approximately 100 mm2 of mouse flank skin, after a dose of 100 Gy which produces transitory moist desquamation. Within 24 hr of such a dose the LI of the irradiated area was essentially zero (0.07 +/- 0.03%), whilst those of the side area and of the control area were 15.0 +/- 2.6% and 21.4 +/- 2.7%, respectively. The LI of the side and the control areas then fell within 3-5 days to approximately 4% and approximately 2% respectively, whilst that of the irradiated area rose rapidly to a peak value of 30.2 +/- 1.7% at 10 days post-irradiation. There was a 20% reduction in the diameter of the area with detectable radiation damage within 5 days, and this is primarily due to cell proliferation and migration from the unirradiated margins of the field. In contrast, between days 10 and 20 the major source of repopulation is probably derived from local migration and proliferation of surviving hair follicle basal cells within the irradiated field.

本研究观察了100 Gy剂量的锶-90/钇-90 β照射约100 mm2小鼠侧腹皮肤后,滤泡间DNA标记指数(LI)随时间的变化,该剂量会产生短暂的湿脱皮。在该剂量24小时内,照射区LI基本为零(0.07 +/- 0.03%),而侧区和对照区LI分别为15.0 +/- 2.6%和21.4 +/- 2.7%。侧面和对照区的LI在3-5天内分别下降至约4%和约2%,而照射区LI在照射后10天迅速上升至30.2 +/- 1.7%的峰值。在5天内,可检测到辐射损伤的区域直径缩小了20%,这主要是由于细胞从未受辐射的区域边缘增殖和迁移。相比之下,在第10天至第20天之间,重新种群的主要来源可能是在辐照场内幸存的毛囊基底细胞的局部迁移和增殖。
{"title":"Changes in the DNA labelling index after beta irradiation of the mouse epidermis.","authors":"L S Hansen,&nbsp;J E Coggle,&nbsp;J Wells,&nbsp;M W Charles","doi":"10.1111/j.1365-2184.1984.tb00606.x","DOIUrl":"https://doi.org/10.1111/j.1365-2184.1984.tb00606.x","url":null,"abstract":"<p><p>This study looked at the changes in the interfollicular DNA labelling index (LI) with time after strontium-90/yttrium-90 beta irradiation of approximately 100 mm2 of mouse flank skin, after a dose of 100 Gy which produces transitory moist desquamation. Within 24 hr of such a dose the LI of the irradiated area was essentially zero (0.07 +/- 0.03%), whilst those of the side area and of the control area were 15.0 +/- 2.6% and 21.4 +/- 2.7%, respectively. The LI of the side and the control areas then fell within 3-5 days to approximately 4% and approximately 2% respectively, whilst that of the irradiated area rose rapidly to a peak value of 30.2 +/- 1.7% at 10 days post-irradiation. There was a 20% reduction in the diameter of the area with detectable radiation damage within 5 days, and this is primarily due to cell proliferation and migration from the unirradiated margins of the field. In contrast, between days 10 and 20 the major source of repopulation is probably derived from local migration and proliferation of surviving hair follicle basal cells within the irradiated field.</p>","PeriodicalId":75682,"journal":{"name":"Cell and tissue kinetics","volume":"17 5","pages":"475-81"},"PeriodicalIF":0.0,"publicationDate":"1984-09-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1111/j.1365-2184.1984.tb00606.x","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"17526931","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
引用次数: 0
Can nocodazole, an inhibitor of microtubule formation, be used to synchronize mammalian cells? Accumulation of cells in mitosis studied by two parametric flow cytometry using acridine orange and by DNA distribution analysis. nocodazole是一种微管形成抑制剂,可以用于同步哺乳动物细胞吗?用吖啶橙双参数流式细胞术和DNA分布分析研究了有丝分裂过程中细胞的积累。
Pub Date : 1984-01-01
M Nüsse, H J Egner

Nocodazole, a temporary inhibitor of microtubule formation, has been used to partly synchronize Ehrlich ascites tumour cells growing in suspension. The gradual entry of cells into mitosis and into the next cell cycle without division during drug treatment has been studied by flow cytometric determination of mitotic cells, analysing red and green fluorescence after low pH treatment and acridine orange staining. Determination of the mitotic index (MI) by this method has been combined with DNA distribution analysis to measure cell-cycle phase durations in asynchronous populations growing in the presence of the drug. With synchronized cells, it was shown that in the concentration range 0.4-4.0 micrograms/l, cells could only be arrested in mitosis for about 7 hr and at 0.04 microgram/ml, for about 5 hr. After these time intervals, the DNA content in nocodazole-blocked cells was found to be increased, and, in parallel, the ratio of red and green fluorescence was found to have changed, showing entry of cells into a next cell cycle without division (polyploidization). It was therefore only possible to partially synchronize an asynchronous population by nocodazole. However, a presynchronized population, e.g. selected G1 cells or metabolically blocked G1/S cells, were readily and without harmful effect resynchronized in M phase by a short treatment (0.4 microgram/ml, 3-4 hr) with nocodazole; after removal of the drug, cells divided and progressed in a highly synchronized fashion through the next cell cycle.

诺可达唑是一种微管形成的临时抑制剂,已被用于部分同步埃利希腹水肿瘤细胞在悬浮中生长。通过流式细胞术测定有丝分裂细胞,分析低pH处理后的红色和绿色荧光以及吖啶橙染色,研究了药物治疗期间细胞逐渐进入有丝分裂并进入下一个细胞周期而不分裂。通过这种方法测定有丝分裂指数(MI)与DNA分布分析相结合,以测量在药物存在下生长的异步种群的细胞周期期持续时间。对于同步细胞,结果表明,在0.4-4.0微克/l浓度范围内,细胞只能在有丝分裂中停留约7小时,在0.04微克/ml浓度范围内,细胞只能在有丝分裂中停留约5小时。在这些时间间隔之后,发现诺可达唑阻断细胞中的DNA含量增加,同时发现红色和绿色荧光的比例发生了变化,表明细胞进入下一个细胞周期而没有分裂(多倍体化)。因此,诺可达唑只能使非同步种群部分同步。然而,预先同步的细胞群,例如选定的G1细胞或代谢阻断的G1/S细胞,在M期通过短时间(0.4微克/毫升,3-4小时)用诺可唑重新同步,容易且无有害影响;在去除药物后,细胞分裂并以高度同步的方式进入下一个细胞周期。
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引用次数: 0
Proliferation kinetics of rabbit articular chondrocytes in primary culture and at the first passage. 兔关节软骨细胞原代培养和第一代增殖动力学。
Pub Date : 1983-11-01
X Ronot, C Hecquet, P Jaffray, M Guiguet, M Adolphe, J Fontagne, P Lechat

The in-vitro proliferation kinetics of young rabbit articular chondrocytes were compared in primary culture and at the first passage. The growth curves labelling and mitotic indices, percentage labelled mitosis (PLM) curves and DNA content distributions by flow-microfluorometric analysis during a 7-day growth period were determined in both cases. The length of the cell cycle and the doubling time calculated from the exponential part of the growth curve were quite similar: Tc = 19 hr and Td = 20 hr for the primary culture, Tc = 17 X 3 hr and Td = 20 hr for the first passage. However, the growth curve and the DNA distribution during the 7-day period showed some differences. The duration of the lag period studied by the growth curve was longer in the primary culture than at the first passage. This phenomenon was also observed using the FCM analysis. The growth fraction determination on the second day of culture was in accordance with the lower proliferation capacity of the cells in primary culture. These data suggest that it would be better to study growth kinetics and drug modifications in articular chondrocytes at the first passage than in primary culture.

比较了幼兔关节软骨细胞原代培养和一次传代时的体外增殖动力学。用流动微荧光法测定两组细胞生长7 d期间的生长曲线标记和有丝分裂指数、百分比标记有丝分裂(PLM)曲线和DNA含量分布。由生长曲线指数部分计算的细胞周期长度和倍增时间非常相似:原代培养Tc = 19小时,Td = 20小时,第一代培养Tc = 17 × 3小时,Td = 20小时。但7 d内的生长曲线和DNA分布存在一定差异。生长曲线研究的滞后时间在原代培养中比在第一次传代中更长。FCM分析也观察到这一现象。培养第2天的生长分数测定与原代培养细胞增殖能力较低一致。这些数据表明,在第一次传代时研究关节软骨细胞的生长动力学和药物修饰比在原代培养中更好。
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引用次数: 0
Cell loss from viable and necrotic tumour regions measured by 125I-UdR. 125I-UdR测量存活和坏死肿瘤区域的细胞损失。
Pub Date : 1983-11-01
R Porschen, W Porschen, H Mühlensiepen, L E Feinendegen

Loss of cells from vital and necrotic areas of the syngeneic mammary adenocarcinoma EO 771 in male C57 BL/6J mice may be measured by use of 125I-labelled 5-iodo-2'-deoxyuridine (125I-UdR). Later than 50 hr after an intraperitoneal injection of 20 muCi 125I-UdR the incorporated activity of the entire tumour was externally measured and found to decrease with time after injection. The injected amount was neither chemo- nor radiotoxic. By injecting the vital dye 'light green', unstained necrotic and stained viable regions were separately excised and measured for loss of activity throughout the natural development of the labelled tumour. With the appearance of necrotic regions, labelled viable cells became necrotic, and activity was slowly eliminated. With increasing proportions of necrosis during tumour growth, the rate of loss of activity of the whole tumour decreased. Loss of activity from viable tumour regions reflected cell death and exceeded the loss rates of the whole tumour by a factor of 2 to 3. The data show that loss of activity from the whole tumour results from a superposition of different elimination rates of viable and necrotic tumour regions and is not an immediate consequence of cell death in the course of undisturbed tumour development.

使用125i标记的5-碘-2'-脱氧尿苷(125I-UdR)可以测量雄性C57 BL/6J小鼠同基因乳腺腺癌eo771的重要和坏死区域的细胞损失。在腹腔注射20 muCi 125I-UdR 50小时后,外部测量整个肿瘤的合并活性,发现注射后随时间减少。注射量既没有化学毒性,也没有放射性毒性。通过注射活性染料“浅绿色”,分别切除未染色的坏死区域和染色的活区,并测量标记肿瘤在整个自然发展过程中的活动损失。随着坏死区域的出现,标记活细胞逐渐坏死,活性逐渐消失。随着肿瘤生长过程中坏死比例的增加,整个肿瘤的活动丧失率降低。活肿瘤区域的活性丧失反映了细胞死亡,并且超过了整个肿瘤的损失率2到3倍。数据表明,整个肿瘤的活性丧失是由活的和坏死的肿瘤区域的不同消除率的叠加造成的,并不是在未受干扰的肿瘤发展过程中细胞死亡的直接后果。
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引用次数: 0
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Cell and tissue kinetics
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