摘要:

AbstractWith the use of cloned helper‐independent Friend leukemia virus (F‐MuLV), we have induced a high incidence (∼︁ 70%) of myelomonocytic leukemia in mice resistant (Fv‐6rrorFv‐6rs) to erythroleukemia induction by this virus. The spleen cells from these mice (DBA/2 or BALB/c × DBA/2) were found to contain a high level of progenitor cells capable of forming granulocytic and macrophage colonies (CFU‐GM). These CFU‐GM, however, were different from those in the speens of uninfected mice, as they were either very sensitive to or independent of conditioned medium. No erythroid progenitor bursts (BFU‐E) or precursor (CFU‐E) cells were detected in the speens of these diseased animals. If these mice with myelomonocytic leukemia were kept alive by transfusion of red blood cells from uninfected mice, tumorigenic cell lines, capable of being transplanted, into adult mice can be isolated. Three such cell lines TTA‐1, TTA‐3, and TTA‐9 have been established, and they retain their morphology of monocytes and macrophages as well as being positive for the monocyte‐specific stain α‐naphtyl‐acetate esterase. These myelomonocytic leukemia cell lines can also be induced in culture by speen cell‐conditioned medium to differentiate into macrophages. Other conditioned media such as L‐cell‐conditioned medium, phytophemmaglutinin‐stimulated leukocyte‐conditioned medium, and WEHI‐3 conditioned medium were less effective in their abilities to stimulate differentiatio

ISSN:0021-9541    

DOI:10.1002/jcp.1041170302

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1983

数据来源: WILEY

摘要:

AbstractThe effect of lithium on the growth of mammary epithelial cells from adult virgin and midpregnant BALB/c or BALB/cfC3H mice was tested in a serum‐free collagen gel culture system. The serum‐free medium consisted of a 1:1 mixture of Ham's F12 and Dulbecco's Modified Eagle's medium supplemented with insulin, transferrin, cholera toxin, epidermal growth factor (EGF), and bovine serum albumin fraction V (BSA V). A multifold increase in cell number occurred during 10–12 days of culture in this medium. In dose‐response studies in which the concentration of each component of this serum‐free medium was varied in turn, the addition of LiCL (10 mM) enhanced growth at most concentrations of each factor. However, LiCL could not enhance growth in the absence of insulin or BSA V, but could replace EGF. The optimal concentration of LiCl was 5–10 mM; higher concentrations (20–80 mM) were toxic. KCl (1–10 mM) when added to the serum‐free medium slightly stimulated growth; the addition of NaCl to the medium had little effect on growth. LiCl did not enhance the growth of cells from spontaneous mammary tumors o

ISSN:0021-9541    

DOI:10.1002/jcp.1041170303

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1983

数据来源: WILEY

摘要:

AbstractThrombin binding to formaldehyde‐fixed mouse embryo (ME) cells was visualized by indirect immunofluorescence as a dot‐like pattern with dots of approximately 500 nm diameter located over the entire cell surface. Experiments comparing the binding of125I‐thrombin and dot appearance on parallel cultures indicate that the immunofluorescent pattern is specific for thrombin‐binding to high‐affinity receptors. Similar patterns were observed on cells fixed in ethanol or glutaraldehyde prior to thrombin binding and on cells maintained at 4°C. These patterns were also observed in a number of established cell lines. Thus, thrombin receptors may be clustered prior to thrombin binding on all cells with these receptors. Comparing the amount of125I‐thrombin bound to CHO cells with the number of fluorescent dots per cell indicated that each dot represents a cluster of over 1000 receptors. On ME cells, the number of thrombin receptor clusters per cell ranged from fewer than 50 to over 5,000. Based on previous studies, this indicates that on ME cells each cluster contains an average of approximately 200 thrombin binding sites with some cells having few, if any, receptors and others having more than a million recept

ISSN:0021-9541    

DOI:10.1002/jcp.1041170304

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1983

数据来源: WILEY

摘要:

AbstractSeveral investigators have described hemopoietic colonies expressing multilineage differentiation in culture. We recently identified a class of murine hemopoletic progenitors which form blast cell colonies with very high replating efficiencies. In order to clarify further the relationship between progenitors for blast cell colonies and progenitors for the multilineage hemopoietic colonies in culture, we carried out analyses of kinetic and differentiation properties of murine blast cell colonies. Serial observations of the development of blast cell colonies into multilineage (and single lineage) colonies in cultures of spleen cells obtained from 5‐fluorouracil (5‐FU)‐treated mice confirmed the transitional nature of the murine blast cell colonies. The data also suggested that the early pluripotent progenitors are in G0for variable periods, and that when triggered into cell cycle, they proliferate at relatively constant doubling rates during the early stages of differentiation. The notion that some of the pluripotent progenitors are in G0was also supported by long‐term thymidine suicide studies in which spleen cells were exposed to3H‐thymidine with high specific activity for 5 days in culture, washed, and assayed for surviving progenitors. Comparison of replating abilities of day‐7 and day‐16 blast cell colonies from normal as well as 5‐FU‐treated mice indicated that some of the day‐7 blast cell colonies are derived from maturer populations of progenitors which are sensitive to 5‐FU. In contrast, progenitors for the day‐16 blast cell colonies are dormant in cell cycle and were not affected by 5‐FU treatment. Previously we reported that progenitors for day‐16 blast cell colonies have a significant capacity for self‐renewal. These observations suggest the hypothesis that the capability for self‐renewal is accompanied by long periods of G0, and that once commitment to differentiation takes place, t

ISSN:0021-9541    

DOI:10.1002/jcp.1041170305

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1983

数据来源: WILEY

摘要:

AbstractThe plant lectin concanavalin A (Con A), at concentrations of 5–200 μg/ml, induced a twofold to fivefold increase in spontaneous beat rate of cultured aggregates of ventricular cells from seven‐day chick embryos. This response was time, dose, and temperature dependent and was accompanied by a decrease in transmembrane potential. It could be blocked or reversed by α‐methyl‐D‐mannoside but was not reversed by dilution alone. Binding of the lectin occurred in the cold, but a temperature‐dependent process was also necessary to produce the response. Divalent (succinyl) Con A did not cause a beat rate increase. Whole heart aggregates responded similarly but less intensely than ventricular aggregates. Atrial aggregates, and whole heart aggregates treated with 5 μg/ml of Con A, produced a biphasic chronotropic response, first decreasing then increasing their beat rates. These results suggest that saccharide‐bearing macromolecules on the heart cell surface play a role in regulating spont

ISSN:0021-9541    

DOI:10.1002/jcp.1041170306

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1983

数据来源: WILEY

摘要:

AbstractCardiac cells obtained from neonatal rat heart contain a mixed population of cell types that can be enriched in culture in either myocytes or fibroblast‐like cells. A metabolic comparison of mixed heart cell cultures with enriched cultures of the same age‐in‐culture and initial cell density showed that mixed cultures used glucose more rapidly than either enriched myocytes or fibroblasts. Mixed cultures were shown to respond to deprivation of insulin and of serum with decreases in the rate of glucose usage and decreases in the protein content of cells, whereas enriched cultures did not respond in the expected manner to insulin deprivation. Mixed, 11‐day‐old cells also exhibited greater increases in cellular protein and greater resistance to the stress of starvation than enriched cultures. Palmitate usage, however, was similar in all cultures examined. We conclude that mixed cultures may serve as a better model system to study cardiac metabolism and to monitor the effects of drugs and hormones on the neonatal myocardium. In addition, it is clear from our results that myocytes and fibroblastic‐like cells coexist in a metabolically functiona

ISSN:0021-9541    

DOI:10.1002/jcp.1041170307

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1983

数据来源: WILEY

摘要:

AbstractA new, gentle technique has been developed for washing of the retinal interphotoreceptor space (IPS) to obtain soluble components of the extracellular matrix (ECM). Using this method, we have determined that the major soluble coustituent of monkey IPS is a 146,000 Mrglycoprotein, which binds [3H]retinol, sediments on sucrose gradients at 7S and has an Rfof 0.42 on native gel electrophoresis. Using size‐exclusion high performance liquid chromatography, the apparent molecular weight of the native protein was calculated to be 250,000 daltons. In contrast to previous studies, no 15,000‐dalton cellular retinol‐binding protein (CRBP) or 33,000‐dalton cellular retinaldehydebinding protein (CRALBP) was observed in the IPS wash, indicating that these proteins are probably not involved in retinol transport between retina and pigment epithelium (PE). In the supernatant fraction of retinal homogenates that contains soluble intracellular proteins as well as extracellular constituents, the 146,000 Mrprotein was closely associated with a 93,000 Mrprotein that could be separated on SDS‐gel electrophoresis; the 93,000 Mrprotein was not found in the IPS wash. The 146,000 Mrinterphotoreceptor retinol‐binding protein (IRBP) may function in extracellular retinol transport

ISSN:0021-9541    

DOI:10.1002/jcp.1041170308

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1983

数据来源: WILEY

摘要:

AbstractThe kinetics of aging of normal human diploid brain cells in culture have been determined using the miniclone technique in which cells are cloned in the presence of a large number of other cells. The miniclone technique records the behaviour of every viable cell in the sample, not merely those cells capable of forming visible clones. This technique permits the direct measurement of the reproductive potential of individual cells growing in buik culture and of the dispersion of the sizes of colonies generated by dividing cells. The fraction of cells that are able to divide declines smoothly and continuously from the beginning of in vitro cultures of human glial cells. There is a broad distribution of colony sizes; even at the earliest passages there are significant numbers of small colonies. With increasing age of the culture there is a shift in the distribution, so that fewer large colonies and more small colonies occur. The distribution of intermitotic times is almost identical in young and middle‐aged cultures. Our data seem to exclude quite positively any description in terms of a catastrophe or any abrupt change in the population. On the contrary, the decline in reproductive potential may be described adequately either as a linear change with time, or as predicted by the mortality theory of Shall and Stein (1979), in which the single constant, gamma, describes the change in reproductive potential over the entire lifetim

ISSN:0021-9541    

DOI:10.1002/jcp.1041170309

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1983

数据来源: WILEY

摘要:

AbstractVimentin‐positive, desmin‐negative cells were established in culture from the nodule and from apparently normal palmar aponeurosis of a patient with Dupuytren's disease and compared with normal human embryonic and adult fibroblasts or sarcomatous cells. Cells from the nodule display in vitro biological properties that are intermediate between those expressed by normal fibroblasts and sarcoma cells or cells from the nodule transformed with SV40 virus. Thus, they represent an interesting in vitro model of partially transformed human cells. This behavior is not evolutive and justifies the classification of Dupuytren's disease among the benign mesenchymal tumors. The production of high level of plasminogen activator probably explains the local reactive pathology, and could act as a mitogenic stimulus for the proliferation of the nodule itself. Cultures derived from the apparently normal palmar aponeurosis show some but not all the abnormal growth properties of cells from nodules; this may help to explain the onset of local recurrences. Our results suggest that Dupuytren's disease is not strictly local and limited to the nodules, but affects, at least partially, the whole aponeurosis. Dupuytren's nodules could be considered as a model of tumor progression in a benign situat

ISSN:0021-9541    

DOI:10.1002/jcp.1041170310

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1983

数据来源: WILEY

摘要:

AbstractSerially cultivated with 3T3 feeder layer support as colonies of stratified squamous epithelium, rat epidermal and esophageal epithelial cells were readily distinguishable by three critera. First, the epidermal colonies, exhibiting extensive piling up of squarnes in the centers, were more stratified than esophageal colonies. Second, in sparse culture 70 to 90% of the esophageal cells but as few as 1 to 5% of the epidermal cells were competent in cross‐linked envelope formation upon treatment with the ionophore X537A. After reaching confluence, up to 90% of the cells of both types formed envelopes upon ionophore treatment. Third, epidermal cells in suspension culture reached maximal levels of spontaneously cross‐linked envelopes in 1 day or less, while esophageal cells required about 4 days in suspension to reach maximal levels. A reproducible finding with both cell types was that initial colony‐forming efficiencies of less than 1% increased to about 40% upon serial passage with consequent derivation of continuous lines. Sparse cultures of esophageal cells with high colony‐forming ability retained a high degree of envelope competence (70 to 90%), indicating these two properties are not mutually exclusive. The derived lines exhibited reduced dependence upon feeder layer support at clonal density, but in suspension culture the cells did not grow and lost colony‐forming ability with a half‐time of several hours. We conclude that cells from these keratinized rat epithelia exhibit intrinsic differences in culture and become continuous lines expressing characteristic regulation of envelope competence and loss of germinative capability in

ISSN:0021-9541    

DOI:10.1002/jcp.1041170311

出版商:Wiley Subscription Services, Inc., A Wiley Company    

年代:1983

数据来源: WILEY