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

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)

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

Journal of Cell Science ◽

10.1242/jcs.86.1.83 ◽

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.

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Steroid responsiveness of the human cell line NHIK 3025

Acta Endocrinologica ◽

10.1530/acta.0.0970551 ◽

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.

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

Parasitology ◽

10.1017/s0031182000061461 ◽

1989 ◽

Vol 98 (3) ◽

pp. 387-393 ◽

Cited By ~ 19

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.

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

10.1101/2021.09.14.460267 ◽

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.

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Biosynthesis of proteoglycans and hyaluronate in rabbit corneal fibroblast cultures. Variation with age of the cell line and effect of foetal calf serum

Experimental Eye Research ◽

10.1016/s0014-4835(81)80021-8 ◽

1981 ◽

Vol 32 (4) ◽

pp. 419-433 ◽

Cited By ~ 16

Author(s):

Inger Marie S. Dahl

Keyword(s):

Cell Line ◽

Calf Serum ◽

Foetal Calf Serum ◽

Corneal Fibroblast ◽

Fibroblast Cultures

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A New Human Breast Tumor Cell Line

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100115560 ◽

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.

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Fish kidney cell line in response to heat shock

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100123921 ◽

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.

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