scholarly journals An improved culture protocol for the differentiation and survival of human promyelocytic leukaemia PLB-985 cell-line into mature neutrophil-like granulocytes

2021 ◽  
Author(s):  
Shehu Shaayau ◽  
Andrew L Cross ◽  
Helen L Wright ◽  
Steven W Edwards
Keyword(s):  
Cell Line ◽  
Genetic Manipulation ◽  
Ex Vivo ◽  
Calf Serum ◽  
Human Neutrophils ◽  
Foetal Calf Serum ◽  
Differentiated Cells ◽  
Mature Neutrophil ◽  
Blood Neutrophils ◽  
Culture Protocol

Circulating blood neutrophils are short-lived, lack proliferation capacity and cannot be transfected in vitro to express exogenous genes or proteins. These properties have made the ex vivo genetic manipulation of neutrophils challenging and hindered biochemical and molecular studies investigating the function of specific genes and proteins. Improved methodology for differentiating cell lines into mature neutrophil-like phenotypes, with similar morphological and functional properties to blood neutrophils would, therefore, be an important tool to probe the molecular properties of mature cells. The PLB-985 cell line was cultured in RPMI-1640 medium supplemented foetal calf serum (FCS) and penicillin/streptomycin. For induction of differentiation into neutrophil-like cells, the medium was supplemented with sodium pyruvate, N, N-dimethylformamide (DMF) and all-trans retinoic acid (ATRA), FCS and penicillin/streptomycin. The cytokines G-CSF and GM-CSF were used to enhance differentiation, prolong viability and delay the progression of the differentiated cells into apoptosis. The modified culture protocol and conditions induced PLB-985 cells to differentiate into mature, neutrophil-like granulocytes that resembled the morphology of mature blood neutrophils as evident by acquisition of a multi-lobed nucleus and granulated cytoplasm. These modified culture conditions resulted in enhanced differentiation into neutrophil-like cells and the apoptosis of these differentiated cells was delayed by supplementation with cytokines. This experimental system should be useful for studies probing the function of specific genes and proteins in human neutrophils.

scholarly journals Control by fibroblast growth factor of differentiation in the BC3H1 muscle cell line.

1985 ◽  
Vol 100 (5) ◽  
pp. 1540-1547 ◽  
Author(s):  
B Lathrop ◽  
E Olson ◽  
L Glaser
Keyword(s):  
Growth Factor ◽  
Cell Growth ◽  
Fibroblast Growth Factor ◽  
Cell Line ◽  
Calf Serum ◽  
Specific Protein ◽  
Fibroblast Growth ◽  
Mouse Muscle ◽  
Differentiated Cells ◽  
High Concentrations

The regulation of creatine phosphokinase (CPK) expression by polypeptide growth factors has been examined in the clonal mouse muscle BC3H1 cell line. After arrest of cell growth by exposure to low concentrations of serum, BC3H1 cells accumulate high levels of muscle-specific proteins including CPK. The induction of this enzyme is reversible in the presence of high concentrations of fetal calf serum, which cause quiescent, differentiated cells to reenter the cell cycle. Under these conditions, the rate of CPK synthesis is drastically reduced. We show in the present communication that either pituitary-derived fibroblast growth factor (FGF) or brain-derived FGF are as effective as serum in repressing the synthesis of CPK when added to quiescent, differentiated cells. The decrease in the rate of synthesis of CPK occurs within 22 h after the addition of pituitary FGF to the cells. Pituitary FGF had very little effect, if any, on the rate CPK degradation. The overall rate of protein synthesis and the pattern of synthesis of the major polypeptides made by these cells was not altered by the addition of FGF. Although pituitary FGF was mitogenic for BC3H1 cells, the rate of cell growth was not absolutely correlated with the extent of repression of CPK. Brain-derived FGF fully repressed CPK induction under conditions where it showed no significant mitogenic activity. These results show that the expression of a muscle-specific protein, CPK, can be controlled by a single defined polypeptide growth factor in fully differentiated cultures, and that initiation of cell division is not required for their regulation to take place.

Substitution of transferrin by FeCl3 in the development of a low foetal calf serum concentration medium for KB-26.5 hybridoma cell line

1993 ◽  
Vol 13 (2) ◽  
pp. 133-141 ◽  
Author(s):  
Bo Damgaard ◽  
Anna Sanfeliu ◽  
Jordi Joan Cair� ◽  
Carles Casas ◽  
Carles Sol� ◽  
...  
Keyword(s):  
Serum Concentration ◽  
Cell Line ◽  
Calf Serum ◽  
Foetal Calf Serum

Endocrine and mucous differentiation by a cloned human rectal adenocarcinoma cell line (HRA-19) in vitro: inhibition by TGF-beta 1

1994 ◽  
Vol 107 (4) ◽  
pp. 1041-1046
Author(s):  
S.C. Kirkland ◽  
K. Henderson
Keyword(s):  
Cell Line ◽  
Endocrine Cells ◽  
Calf Serum ◽  
Rectal Adenocarcinoma ◽  
Model Systems ◽  
Foetal Calf Serum ◽  
Mucous Cells ◽  
Extrinsic Factors ◽  
Colorectal Epithelium

Colorectal epithelium is composed of absorptive, mucous and endocrine cells, all of which are considered to arise from a common stem cell located in the crypt base. However, the factors controlling the commitment to differentiate are poorly understood. This is partly due to the lack of in vitro model systems for the study of differentiation in colorectal epithelium. The HRA-19 cell line, established from a human rectal adenocarcinoma, has been shown to have multipotential characteristics with cloned HRA-19 cells able to differentiate into absorptive, mucous and endocrine cells when grown as xenografts. The lack of such differentiated cells in HRA-19 monolayers in vitro suggests that differentiation is controlled by extracellular matrix, stromal cells and/or soluble factors. Such observations show that differentiation in HRA-19 cells can be controlled by extrinsic factors and therefore provide a model system for studying control of differentiation in colorectal epithelium. Unfortunately, the restriction of differentiation to xenografts of the cell line limits the degree to which this differentiation can be manipulated. In this study, the possibility that HRA-19 cells could be induced to differentiate in vitro under appropriate conditions has been investigated. Endocrine and mucous cells were identified by immunocytochemistry with differentiation-related antibodies and histology of monolayers. Preconfluent HRA-19 cells grown in 10% foetal calf serum formed a well polarised monolayer with apical tight junctions and sparse microvilli, but cells with mucous or endocrine phenotypes were only very occasionally observed. However, endocrine and mucous cells could reproducibly be demonstrated in postconfluent monolayers grown in 1% foetal calf serum.(ABSTRACT TRUNCATED AT 250 WORDS)

Characterization of a spontaneously transformed pulmonary embryonic rat (PER) epithelial cell line

1986 ◽  
Vol 86 (1) ◽  
pp. 83-93
Author(s):  
M. Paye ◽  
C. Etievant ◽  
F. Michiels ◽  
D. Pierard ◽  
B. Nusgens ◽  
...  
Keyword(s):  
Growth Factor ◽  
Epithelial Cell ◽  
Cell Line ◽  
Calf Serum ◽  
Maximum Growth ◽  
Soft Agar ◽  
Maximum Growth Rate ◽  
Epithelial Cell Line ◽  
Foetal Calf Serum

A spontaneously transformed pulmonary embryonic rat epithelial cell line (PER) is described in terms of growth, tumorigenicity, growth factor responsiveness and biosynthetic capacity. At low-passage subcultures, PER cells grew as a monolayer and did not form colonies in soft agar. After long-term subcultivation, they lost contact inhibition, became anchorage-independent and formed tumours in nude mice. Low concentrations of foetal calf serum permit the maximum growth rate. The multiplication and metabolic activity, assessed by 2-deoxy-D-glucose uptake, was significantly stimulated by growth factors. PER cells synthesized collagen types I, III, IV and V, laminin and fibronectin, and organized a pericellular matrix made up of only basement membrane components (type IV collagen and laminin) and fibronectin. These data enabled us to define PER cells as a transformed epithelial cell line evolving towards malignancy with long-term subcultivation. These cells appeared to be a valuable tool in studies of tumour cell-matrix interactions and regulation of growth factor receptors in tumorigenesis.

Steroid responsiveness of the human cell line NHIK 3025

1981 ◽  
Vol 97 (4) ◽  
pp. 551-558
Author(s):  
Kjetill Østgaard ◽  
Einar Wibe ◽  
Kristen B. Eik-Nes
Keyword(s):  
Cell Cycle ◽  
Cell Proliferation ◽  
Androgen Receptor ◽  
Cell Growth ◽  
Cell Line ◽  
Human Cell ◽  
Calf Serum ◽  
Human Cell Line ◽  
Foetal Calf Serum ◽  
Steroid Responsiveness

Abstract. The human cell line NHIK 3025, derived from a carcinoma of the uterine cervix, contains a glucocorticoid and an androgen receptor. The effect of various natural and synthetic steroid hormones and antihormones on growth rate of these cells was therefore investigated. Cells grown in Eagle's MEM with 10% foetal calf serum exhibited reduced growth when cultured with dexamethasone due to prolongation of the cell cycle. Glucocorticoid anti-inducers like progesterone had no significant effect on cell growth. Methyltrienolone (R 1881) or 5α-dihydrotestosterone did not affect cell proliferation. The reported shortening of the cell cycle by testosterone is probably not directly connected with activation of the androgen receptor present, but possibly dependent on metabolic conversion of testosterone to the more potent growth stimulator 4-androstene-3β, 17β-diol. The effect of several anti-androgens was also studied. The non-steroidal anti-androgens flutamide and SCH 16483 had no significant effect on cell proliferation. It was, however, found that a number of steroid antiandrogens, including R2956, stimulated cell growth. A significant stimulatory effect by R2956 was seen within the first cell generation, 4-androstene-3β,17β-diol had to be present during 2 days, and testosterone for even longer times before a similar effect on cell growth could be obtained.

Cultivation of Blastocrithidia triatomae (Trypanosomatidae) on a cell line of its host Triatoma infestans (Reduviidae)

Parasitology ◽  
1989 ◽  
Vol 98 (3) ◽  
pp. 387-393 ◽  
Author(s):  
Dagmar Reduth ◽  
G. A. Schaub ◽  
Mary Pudney
Keyword(s):  
Cell Line ◽  
Primary Cultures ◽  
Calf Serum ◽  
Host Cells ◽  
Triatoma Infestans ◽  
Foetal Calf Serum ◽  
Tryptose Phosphate Broth ◽  
A Cell ◽  
Vigorous Growth

SUMMARYBlastocrithidia triatomae parasitizes vectors of Chagas' disease and is very difficult to cultivate in conventional media. However, co-cultivation with a cell line of its host Triatoma infestans (TI-32; in Schneider's Drosophila medium supplemented with 20% foetal calf serum and 10% tryptose phosphate broth) led to vigorous growth at 24 or 28 °C without an adaptation phase. More than 60 primary cultures were initiated successfully without any failures. Subcultures could be started immediately at weekly intervals. The doubling time was similar in both the primary cultures (41 h) and the 33rd subculture (39 h). The importance of the reduviid cells for B. triatomae became clear after removal of the insect cells, when multiplication of epimastigotes stopped and mainly cysts were formed. Cysts produced in vitro were infective for reduviids. Scanning electron microscopy showed that B. triatomae attached to the host cells, inserted its flagellum into them and destroyed them.

Effects of oxytocin and vasopressin on the preadipocyte 3T3-F442A cell line

1987 ◽  
Vol 65 (3) ◽  
pp. 211-218 ◽  
Author(s):  
E. Jane Wilson ◽  
Morley D. Hollenberg
Keyword(s):  
Glucose Metabolism ◽  
Cell Line ◽  
Cultured Cells ◽  
Calf Serum ◽  
Fibroblast Cell ◽  
Mouse Fibroblast ◽  
Differentiated Cells ◽  
Mouse Fibroblast Cell Line ◽  
Isolated Adipocytes ◽  
Inhibition Of Differentiation

The 3T3-F442A mouse fibroblast cell line, triggered by factors present in fetal calf serum (FCS), converts either spontaneously or, in the simultaneous presence of FCS and insulin, at an accelerated rate into cells exhibiting the adipocyte phenotype. The effects of the neurohypophysial hormones in differentiated cells on glucose metabolism (glucose oxidation and lipogenesis) were compared with the stimulatory actions of insulin, which had its most pronounced effects in cells differentiated spontaneously with FCS in the absence of insulin. The differentiated 3T3-F442A cells were sensitive to physiological levels of insulin and exhibited manyfold increases in glucose metabolism in response to it. This result demonstrated that these cultured cells respond to insulin, in a manner analogous to freshly isolated adipocytes. In contrast to its insulin-like effects in isolated epididymal adipocytes, oxytocin was not reproducibly able to stimulate glucose metabolism in differentiated 3T3-F442A cells. Vasopressin was similarly inactive. In contrast, both oxytocin and vasopressin blocked adipocyte conversion triggered by FCS, either in the presence or absence of insulin; vasopressin was more potent than oxytocin, indicating that a vasopressin receptor was responsible for the observed inhibition of differentiation. Our work suggests that vasopressin could potentially play a role in the regulation of the adipocyte differentiation process.

A New Human Breast Tumor Cell Line

1975 ◽  
Vol 33 ◽  
pp. 388-389
Author(s):  
R.E. Nordquist ◽  
R.M. Wasik ◽  
P.J. Riggs ◽  
P.L. Munson ◽  
F.B. Schafer
Keyword(s):  
Electron Microscopy ◽  
Cell Line ◽  
Calf Serum ◽  
Tumor Cell Line ◽  
Electron Microscopic Examination ◽  
Epithelial Cell Line ◽  
Electron Microscopic ◽  
Fixed Tissue ◽  
Excised Tissue ◽  
Caucasian Female

An infiltrating ductal cell carcinoma was removed from the breast of a postmenopausal Caucasian female. The excised tissue was divided into three parts; one part for electron microscopy, one part for tissue culture and the remainder frozen for immunological studies.The tissue for culture was minced finely with sterile razor blades and cultured in Falcon flasks containing Eagel's MEM supplemented with 10% heat denatured fetal calf serum. The tissue for electron microscopy was fixed in 6.25% glutaraldehyde in 0.1 M PO4 buffer plus 5% sucrose and postfixed in 1% OsO4 in the same buffer. The fixed tissue was dehydrated in graded ethanol and embedded in Spurr.The tissue which was cultured began to grow out after approximately six weeks and became a continuous epithelial cell line which was designated BOT-2 (Breast Original Tumor). Electron microscopic examination revealed that these cells had epithelial characteristics, i.e. the presence of tonofilaments and well formed desmosomes.

Fish kidney cell line in response to heat shock

1992 ◽  
Vol 50 (1) ◽  
pp. 704-705
Author(s):  
Li-Chu Tung ◽  
Yung-Reui Chen ◽  
Shiu-Nan Chen ◽  
Guang-Hsiung Kuo
Keyword(s):  
Heat Shock ◽  
Cell Line ◽  
Fetal Calf Serum ◽  
Calf Serum ◽  
Uranyl Acetate ◽  
Ultrastructural Changes ◽  
Heat Shock Treatment ◽  
Acanthopagrus Schlegeli ◽  
Cacodylate Buffer ◽  
Fish Kidney

In the present study, the ultrastructural changes of BPK cells, a fibroblast-like cell line, derived from the kidney of juvenile black porgy Acanthopagrus schlegeli, under heat shock treatment are described.The BPK cells were maintained in L-15 medium supplemented with 10% fetal calf serum and 0.15 M NaCl at 28|C2. The heating was carried out in precalibrated water baths. Monolayers of cells, grown on coverslips in parafilm-sealed petri dishes were submerged under water for 30 min at 40|C treatments. Cells were fixed in 2.5% glutaraldehyde in 0.1 M cacodylate buffer supplemented with 6.6% sucrose, postfixed in 1% OsO4 and flat embedded in Spurr’s resin. Silver section were cut parallel to the substratum, stained with uranyl acetate and Reynold’s lead citrate, and examined in a Hitachi H-600 electron microscope at 75 KV.

Sign in / Sign up

Export Citation Format

Share Document