Nutrient factors present in medium conditioned by a human cloned myelomonocytic cell line growth at limiting dilution (original) (raw)

The Growth of Bone Marrow Cells in Liquid Culture

British Journal of Haematology, 1972

Liquid suspension cultures of mouse bone marrow cells at high and low density were prepared in supplemented Eagle's medium containing 10% of a partially purified extract of mouse embryos and pregnant mouse uterus (PMU).

Response of Human Myeloid Leukemia Cells to Various Sources of Colony stimulating Activity and Phytohemagglutinin-conditioned Medium1

The response of human myeloid leukemia cells to various sources of colony-stimulating activity (CSA) and media condi tioned by phytohemagglutinin-stimulated mononuclear cells (PHA-LCM) was investigated in liquid and colony culture. PHA-LCM, placenta-conditioned medium, GCT cell line-conditioned medium, leukocyte-conditioned medium, and partially purified CSA for human and murine cells were tested for ability to support growth of granulocyte-macrophage colonies from adherent celldepleted human bone marrow. This activity was correlated with ability to support leukemia colony growth in methylcellulose, and [3H]thymidine incorporation in liquid culture by normal bone marrow cells, leukemia cells, and the KG-1 myeloid leukemia cell line. For normal cells, growth and liquid culture responses were highly correlated for various sources of CSA (r = 0.92), and addition of data using PHA-LCM changed results only slightly (r = 0.89).

Factors in Human Serum Affecting the Proliferation of Normal and Leukemic Cells1

Serum factor(s) regulate the rate of proliferation of normal and leukemic bone marrow cells. Nine patients with solid tumors and normal marrow function were treated with cyclophosphamide (60.0 mg/kg) on each of 2 successive days. Following aplasia, the granulocytic proliferative fraction of the patients' marrow increased twofold (tritiated thymidine autoradiography). With maturation of the marrow elements, the tritiated thymidine indices fell below pretreatment values. Serial serum samples obtained from these patients affected the incorporation of tritiated thymidine into DNA in primary cultures of normal bone marrows. In relation to pretreatment sera, samples obtained during the proliferative phase caused a twofold increase in tritiated thymidine incorporation by target cells, and sera obtained during the marrow maturation phase caused a twofold decrease. Identical results were obtained when these sera were incubated with marrows from patients with acute lymphoblastic and acute myelocytic leukemia (> 95% tumor) and when they were assayed with the long-term cultured cell lines.

Clonal growth and self-renewal of human myeloid leukemia cell lines (BRM and DD) in serum-free semi-solid culture

P r i m a r y and secondary colony f o r m a t i o n o f two new h u m a n myeloid leukemia cell lines (BRM and DD) were studied in serum-free semisolid cultures. The results indicate that bovine serum a l b u m i n and transferrin were essential for clonal growth in chemically defined m e d i u m . Insulin contributed only moderately beneficial effects. Initial cell density was also a m a j o r m o d u l a t o r o f plating efficiency. Positive cooperation between the leukemia cells was shown by using autologous conditioned media. This is the first serum-flee culture m e t h o d that allows self-renewal of h u m a n myeloid leukemia cell lines in terms of secondary colony formation in methylcellulose cultures.

Comparative studies on media supplementation with various types of sera in tissue cultures

The present study was conducted to evaluate the growth promoting effect of different types of serum including; fetal calf serum (FCS), calf serum, sheep serum, goat serum, horse serum and African catfish serum on VERO cell cultures. These animal sera are used as nutrient supplements to VERO cell cultures. The evaluation of VERO cell growth was carried out by three techniques; turbid metric measurement, manual counts by hemocytometer and methylene blue dye reduction test. The data obtained by applying the three methods of evaluation revealed that, VERO cells proliferation in tissue culture media containing the different tested sera with different concentrations was variable. FCS had the best growth rate followed by calf serum, goat serum and African cat fish serum. Horse serum and sheep serum had the least growth rates among all of the tested sera. Hence, African cat fish serum can be used as an alternative to overcome the disadvantages of other sera.

Establishment and characterization of a new human cloned myelomonocytic cell line (ZC-1.6)

Experimental Hematology

A human cell line, ZC, derived from the blood of a patient with acute myelomonocytic leukemia, was established and cloned. One of the clones, ZC-1.6, was subsequently characterized. As for its morphology and cytochemistry, ZC-1.6 clone shares a number of features with immature cells of the monocytic or myelocytic lineages. Surface marker analysis shows positivity for 4F2 (100% of the cells), and OKM1 (38%) monoclonal antibodies, and presence of surface HLA-D/DR antigens (100%) and Fc (11%) and complement (C3b) receptors (100%). Functional capabilities of ZC-1.6 cells include adherence, spreading, and phagocytosis of latex and opsonized zymosan particles. Despite its morphological immaturity, the ZC-1.6 clone produces relevant amounts of O2- in the presence of different stimuli (phorbol myristate acetate, opsonized zymosan, or latex particles). The production of O2- by ZC-1.6 cells is the first evidence that reactive oxygen intermediate production may represent an early feature of cells of the myelomonocytic lineage.

Asymmetric cellular responses in primary human myoblasts using sera of different origin and specification

PloS one, 2018

For successful growth and maintenance of primary myogenic cells in vitro, culture medium and addition of sera are the most important factors. At present it is not established as to what extent sera of different origin and composition, supplemented in media or serum-free media conditions influence myoblast function and responses to different stimuli. By assessing markers of proliferation, differentiation/fusion, quiescence, apoptosis and protein synthesis the aim of the current study was to elucidate how primary human myoblasts and myotubes are modulated by different commonly used serum using FCS (foetal calf serum), (CS-FCS charcoal-stripped FCS, a manufacturing process to remove hormones and growth factors from sera), HS (horse serum) as well as in serum free conditions (DMEM). To characterise the biological impact of the different serum, myoblasts were stimulated with Insulin (100 nM) and Vitamin D (100 nM; 1α,25(OH)2D3, 1α,25-Dihydroxycholecalciferol, Calcitriol), two factors wit...

Effects of conditioned medium upon proliferation, deoxynucleotide metabolism, and antimetabolite sensitivity of promyelocytic leukemic cells in vitro

Cancer Research

In order to investigate the interaction of factors from leukocyteconditioned medium with leukemic cells, effects of an ammonium sulfate-precipitated conditioned medium concentrate were tested upon HL-60 cells. This preparation increased tritiated thymidine incorporation into DNA of HL-60 cells markedly, an effect which was found to be attributable in large part to greater thymidine accumulation in the intracellular nucleotide triphosphate pool. A modestly expanded population of DNA synthetic phase cells was also demonstrated by flow cytometry. Similar effects were noted of the K562 and KG-1 cell lines and were also demonstrable with giant cell tumor-conditioned medium. These effects were not demonstrable with a purified preparation of granulocyte-monocyte colony-stimulated factor. Because of the altered pattern of nucleotide metabolism noted, the effect of the conditioned medium concentrate upon 5-fluorouracil sensitiv ity was tested. Following continuous 24-h exposure to 5-fluo rouracil at 5 x 10~6 M, tritiated thymidine incorporation of HL-60 cells increased in parallel with depletion of endogenous thymidylate. Conditioned medium concentrate markedly sensitized cells to this effect of 5-fluorouracil and also increased growth retardation, cytotoxicity, and cell cycle arrest as assessed by flow cytometry. These studies thus demonstrated marked effects of a factor in conditioned medium on deoxynucleotide uptake and metabolism of the HL-60 line. These effects occurred in conjunction with, but were relatively more marked than, effects upon cell cycle distribution and were found to influence chemo therapy sensitivity.