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1.
Karimullah A. Zirvi Darwin O. Chee George J. Hill 《In vitro cellular & developmental biology. Plant》1986,22(7):369-374
Summary Five human tumor cell lines were studied for growth factor requirements and for replication in serum-free media. Of the five
tumor lines HT-29 (colon carcinoma), TWI (melanoma), A-549 (lung carcinoma), Panc-1, (carcinoma of the pancreas) and EJ, (bladder
carcinoma) only HT-29 and TWI grew in the serum-free medium (SFM). In a series of additional experiments, a combination of
transferrin (5 μg/ml), insulin (5 μg/ml), triiodothyronine (2×10−10
M), epidermal growth factor (20 ng/ml), and selenium (5 ng/ml) was added to Chee’s essential medium (CEM) without serum (C-TITES
medium). The C-TITES modification of CEM was found to allow optimal replication of HT-29 and TWI cells. Both HT-29 and TWI
cells have replicated continuously in C-TITES medium for periods of more than 15 mo. These cells replicate with slightly lower
doubling times than in CEM supplemented with 10% fetal bovine serum. Deletion of insulin or transferrin from the C-TITES medium
resulted in cessation of cell growth of HT-29 and TWI. HT-29 assumed a somewhat rounded morphology, whereas TWI grew with
the characteristic fibroblastic morphology in C-TITES medium. Cell line EJ did not grow in C-TITES medium. The other two cell
lines, A-549 and Panc-1, grew in C-TITES medium but their growth rate was much slower than that in SSM. Availability of cell
lines that can be propagated in serum-free, hormone-supplemented medium may aid in the study of the mechanisms by which hormones
influence cell growth.
This work was supported by Veterans Administration Research Awards to two of the authors (Karimullah A. Zirvi and George J.
Hill) and grant no. CA-37138 from the National Cancer Institute. 相似文献
2.
C. H. Uittenbogaart Y. Cantor J. L. Fahey 《In vitro cellular & developmental biology. Plant》1983,19(1):67-71
Summary Human T lymphoid cell lines (MOLT-4f, MOLT-3, HSB-2, CEM) and human B lymphoid cell lines (BJAB, RAJI, WIL-2) were grown longterm
(up to 8 months) in serum-free medium. This medium consisted of Iscove's modified Dulbecco's medium (IMDM), supplemented with
bovine serum albumin (BSA) and transferrin (TF). This serum-free medium containing albumin and transferrin is designated AT-IMDM.
Lipids were not essential. Cell viability remained high, greater than 80%, in the serum-free medium and the cells maintained
their distinctive characteristics. Interleukin-2 (IL-2) production capacity was maintained by the human T lymphoid cell lines
JURKAT-77 and MO in short term culture. This simple medium composed of relatively inexpensive and readily available components
should be useful for studies of lymphoid cell growth and differentiation and lymphoid cell products.
This research was supported by a grant from the National Institutes of Health, CA 12800, and the Concern Foundation of Los
Angeles, and CA 09120 (C. U.) 相似文献
3.
D. Gaillard R. Négrel G. Serrero-Davé C. Cermolacce G. Ailhaud 《In vitro cellular & developmental biology. Plant》1984,20(2):79-88
Summary Ob17 is a clonal cell line isolated from the epididymal fat pad of C57 BL/6J ob/ob mouse that differentiates into adiposelike
cells in serum-supplemented medium. In serum-free medium, this cell line shows increased growth under the addition of insulin,
transferrin, fibroblast growth factor (FGF), and a factor present in extract of rat submaxillary gland (SMGE). This medium
is referred to as 4F. Epidermal growth factor or nerve growth factor cannot replace SMGE, whereas partially purified platelet
extract can substitute for FGF but only partially for SMGE. 4F Medium is able to support the proliferation of cells from other
established preadipocyte clonal lines, HGFu and 3T3-F442A, and also of preadipocyte cells isolated from the stromal-vascular
fraction of rat and mouse adipose tissues. In each case 4F medium is insufficient to support the differentiation of these
cells into adipocytes. Ob17 cells grown and maintained in serum-free hormone-supplemented medium retain the ability to convert
to adiposelike cells after serum addition. This serum requirement for differentiation cannot be substituted by the addition
of growth hormone or of other putative adipogenic factors, or both. The results are discussed with respect to the requirements
for growth and differentiation of the 3T3-L1 and 1246 preadipocyte cell lines previously described.
This work was supported by the “Centre National de la Recherche Scientifique” (Grant 1208-Biochimie du Développement and Grant
4162-Endocrinologie), by the “Ministère de la Recherce et de la Technologie” (Grant 81-L-1322), by the “Fondation pour la
Recherche Médicale,” by NATO (Grant 1704), and by the “Institut National de la Santé et de la Recherche Médicale” (Grant 827006). 相似文献
4.
Thomas B. Shea Eugene S. Berry 《In vitro cellular & developmental biology. Plant》1983,19(11):818-824
Summary An undefined, serum-free medium was developed for use with fish cell cultures. Lactalbumin hydrolyzate, trypticase-soy broth,
Bacto-peptone, dextrose, yeastolate, and polyvinylpyrrolidone were initially combined in 100 ml of distilled H2O, autoclaved, and added to 5% of the final volume of Medium 199. In addition, filter sterilized bovine pancreatic insulin,
glutamine, and nonessential amino acids were added to the medium. The addition of insulin was observed to be unnecessary.
Five fish cell lines [goldfish-derived CAR cells, fathead minnow (FHM) cells, epithelioma papillosum cyprini (EPC) cells,
chinook salmon embryo (CHSE-214) cells, and a new cell line from goldfish air bladders (ABIII)] were all capable of growth
in the serum-free medium at rates equivalent to cells grown in fetal bovine serum (FBS). The morphology of all cell lines,
except CHSE-214 cells, was identical to cells grown in FBS. All cell lines were capable of long-term growth in the serum-free
medium. The CAR, ABIII, EPC, and CHSE-214 cells in the serum-free medium supported the replication of goldfish virus-2 at
levels equivalent to cells grown in FBS. 相似文献
5.
Attachment and growth of anchorage-dependent cells on a novel, charged-surface microcarrier under serum-free conditions 总被引:1,自引:0,他引:1
James Varani Felicia Piel Sean Josephs Ted F. Beals William J. Hillegas 《Cytotechnology》1998,28(1-3):101-109
The present study describes a novel microcarrier substrate consisting of a swellable, copolymer of styrene and divinylbenzene,
derivatized with trimethylamine. The co-polymer trimethylamine microcarriers support the growth of a number of different cell
lines – Madin Darby Bovine Kidney, Madin-Darby Canine Kidney, Vero and Cos-7 – under serum-free conditions, and human diploid
fibroblasts in serum-containing medium. Cells attach to the co- polymer trimethylamine microcarriers as rapidly as they attach
to other charged-surface microcarriers (faster than they attach to collagen-coated polystyrene microcarriers) and spread rapidly
after attachment. All of the cells examined grow to high density on the co- polymer trimethylamine microcarriers. Furthermore,
cells are readily released from the surface after exposure to a solution of trypsin/EDTA. In this respect, the co-polymer
trimethylamine microcarriers are different from other charged-surface microcarriers. Madin-Darby Bovine Kidney cells grown
on this substrate support production of vaccine strain infectious bovine rhinotracheitis virus as readily as on other charged-surface
or collagen-coated microcarriers. Thus, the co-polymer trimethylamine microcarriers combine the positive characteristics of
the currently available charged-surface and adhesion-peptide coated microcarriers in a single product. The viral vaccine production
industry is undergoing considerable change as manufacturers move toward complete, animal product-free culture systems. This
novel substrate should find application in the industry, especially in processes which depend on viable cell recovery.
This revised version was published online in August 2006 with corrections to the Cover Date. 相似文献
6.
Spontaneous establishment and characterization of mouse keratinocyte cell lines in serum-free medium 总被引:6,自引:0,他引:6
M. E. Kaighn R. F. Camalier F. Bertolero U. Saffiotti 《In vitro cellular & developmental biology. Plant》1988,24(8):845-854
Summary Mouse keratinocytes cultures readily develop into established cell lines without undergoing a “crisis” in a newly-developed
serum-free medium, LEP/MK2. LEP/MK2 consists of calcium-free MEM with non-essential amino acids supplemented with 8 factors.
Two lines, MK1 and MKDC4, have been isolated and have now doubled more than 400 and 200 times respectively. In MK1 cells,
Giemsa banding has revealed significant karyotypic changes as early as the 4th passage, leading to a near-tetraploid karyotype
with random loss and gain of individual chromosomes. Minute chromosomes, but no stable markers have been observed. After these
initial changes, examination of cultures at several passage levels has shown that the karyotype has remained essentially stable.
The MKDC4 line, also sub-tetraploid at the 7th passage, had 4 marker chromosomes by the 47th passage. The rapid increase in
chromosome number may have contributed to the “immortalization” of these lines.
The response of these established keratinocyte lines to growth factors and serum-derived inhibitors changed with increasing
passage level. Most notable of these changes were a reduction in the requirement for bovine pituitary extract (an absolute
requirement for growth of secondary MK1 cells) and a decreased sensitivity to serum and serum-derived inhibitors, e.g., transforming
growth factor-β. The established lines, like primary and secondary keratinocytes, remain responsive to calcium-induced terminal
differentiation and are non-tumorigenic in athymic, nude mice. This serum-free system is suitable for transformation studies
with oncogenes and chemical carcinogens.
Editor's Statement Keratinocytes are useful and important models for studies of carcinogenesis and tumor promotion and differentiation.
This paper provides a solid in vitro basis for examination of the cellular endocrinology of these phenomena in vitro, and
implicates TGF beta as a regulator of these cells. 相似文献
7.
Patrick D. Varga Weisz David W. Barnes 《In vitro cellular & developmental biology. Animal》1993,29(6):512-516
Summary Serum-free mouse embryo (SFME) cells are a cell line derived in medium in which serum is replaced with growth factors and
other supplements. These cells display unusual properties: a) they do not lose proliferative potential or show gross chromosomal
aberration upon extended culture, b) they depend on epidermal growth factor (EGF) for survival, and c) they are reversibly
growth inhibited by plasma and serum. Transfection of SFME cells with oncogenes (ras, neu, SV40 T antigen) results in cells that grow in serum-supplemented medium and no longer require EGF for survival. The growth
inhibitory activity of human plasma on SFME cells was investigated. The activity was present in delipidated plasma and was
not dialyzable against 1M acetic acid. The activity precipitated in 33% methanol, bound to concanavalin A-agarose and was retarded by Sephadex G-50
in 200 mM acetic acid. A fifty- to one-hundred-fold purification was achieved, although most of the differential inhibition of untransformed
vs. transformed cells was lost in the course of the purification. 相似文献
8.
Insulin-like growth factor I has similar mitogenic effects to insulin, a growth factor required by most cells in culture,
and it can replace insulin in serum-free formulations for some cells. Chinese Hamster Ovary cells grow well in serum-free
medium with insulin and transferrin as the only exogenous growth factors. An alternative approach to addition of exogenous
growth factors to serum-free medium is transfection of host cells with growth factor-encoding genes, permitting autocrine
growth. Taking this approach, we constructed an IGF-I heterologous gene driven by the cytomegalovirus promoter, introduced
it into Chinese Hamster Ovary cells and examined the growth characteristics of Insulin-like growth factor I-expressing clonal
cells in the absence of the exogenous factor. The transfected cells secreted up to 500 ng/106 cells/day of mature Insulin-like growth factor I into the conditioned medium and as a result they grew autonomously in serum-free
medium containing transferrin as the only added growth factor. This growth-stimulating effect, observed under both small and
large scale culture conditions, was maximal since no further improvement was observed in the presence of exogenous insulin.
This revised version was published online in July 2006 with corrections to the Cover Date. 相似文献
9.
Masami Ogasawara David A. Sirbasku 《In vitro cellular & developmental biology. Plant》1988,24(9):911-920
Summary Growth of the MCF-7, T47D, and ZR-75-1 human breast cancer cells was established in a serum-free defined medium (MOM-1) composed
of a 1∶1 (vol/vol) mixture of Ham's F12 medium and Dulbecco's modified Eagle's medium containing 15 mM HEPES (pH 7.2), 2 mM 1-glutamine, 20 μg/ml glutathione, 10 μg/ml insulin, 10 μg/ml transferrin (Tf), 10 ng/ml selenous acid, 0.3 nM triiodothyronine, 50 μg/ml ethanolamine, 20 ng/ml epidermal, growth factor, 2.0 nM 17β-estradiol, and 1.0 mg/ml bovine serum albumin (BSA). Proliferation in MOM-1 was 50 to 70% of the serum stimulated rate.
Deletion of components from MOM-1 gave a medium (Tf-BSA) containing only HEPES, 10 μg/ml Tf, and 200 μg/ml BSA, which sustained
MCF-7 and T47D cells in a slowly dividing and mitogen responsive state; ZR-75-1 cells required Tf plus 1.0 mg/ml BSA. In Tf-BSA,
insulin and insulin-like growth factor I(IGF-I) were mitogenic with ED50 values of 2 to 3 ng/ml and 30 to 150 pg/ml, respectively, with MCF-7 cells. The T47D cells were responsive to these factors
in Tf-BSA but required 10-fold higher concentrations for ED50. At saturating concentrations, insulin and IGF-I promoted 1.5 to 3.5 cell population doublings over controls in 8 d. At≤ng/ml
concentrations, epidermal growth factor, insulin-like growth factor II, and basic fibroblast growth factor were mitogenic
for human breast cancer cells in Tf-BSA. Mitogen activities in uterus and pituitary extracts were assayed readily in Tf-BSA.
This new method offers a convenient means of comparing the potencies of growth-promoting factors on human breast cancer cells
without interfering activities known to be present in serum.
This work was supported by grants CA-38024 and CA-26617, from the National Cancer Institute, Bethesda, MD, and by American
Cancer Society grant BC-255 and grant 2225 from the Council for Tobacco Research, USA, Inc. 相似文献
10.
H. Muzik M. E. Shea C. C. Lin H. Jamro S. Cassol L. M. Jerry L. Bryant 《In vitro cellular & developmental biology. Plant》1982,18(6):515-524
Summary Attempts were made to adapt human long-term B lymphoblastoid cell lines to prolonged growth in serum-free, chemically defined
media. A newly described medium, which is an enriched modification of Dulbecco’s modified Eagle’s medium containing additional
amino acids and vitamins, was used. The serum is totally replaced by albumin, transferrin, and soybean lipid. The cell lines
were all adaptable from RPMI 1640 over a period of time during which the 10% fetal bovine serum (FBS) concentration was reduced
and then eliminated in successive steps. After 3 to 6 wk minor alterations in cell shape and adhesion were noted without significant
histological changes. Growth characteristics were comparable in the new medium provided a double initial inoculum was used.
A panel of cell surface markers, including surface immunoglobulins, Ia antigens, Fc and complement receptors, and T and B
erythrocyte rosettes, all showed no altered expression. Molecular genotyping of Ia antigens was carried out by 2-D gel electrophoresis.
The antigens showed their full polymorphism without change and were shed into the new culture medium without alteration. Chromosome
analysis was performed on Q-banded karyotypes from one of the lines and showed no alteration resulting from the change to
serum-free conditions. Thus long-term B lymphoblastoid cell lines can be adapted to prolonged growth in serum-free medium.
This will facilitate the assay and isolation of cell products regulating lymphocyte function and the identification and characterization
of cell surface molecules free of interference from undefined serum components.
This work is supported by grants from the Medical Research Council of Canada, the National Cancer Institute of Cancer, and
the Alberta Heritage Fund. 相似文献
11.
Terry Golombick R. Dansey W. R. Bezwoda Jennifer Rosendorff 《In vitro cellular & developmental biology. Plant》1990,26(5):447-454
Summary The establishment, growth, and characterization of two new continuously growing human ovarian cancer cell lines (UWOV1 and
UWOV2) as, well as a subline (UWOV2, Sf) grown in chemically defined, serum-free medium are described. The cell lines were
derived from ascitic tumors of two patients suffering from cystadenocarcinomas of the ovary. Both UWOV1 and UWOV2 lines grow
in anchorage-dependent fashion as monolayers, whereas UWOV2 (Sf) forms multilayered domelike structures. Cytogenetic studies
revealed nonrandom abnormalities involving chromosomes 1 and 11 in all three cell lines. Secretion of soluble collagen was
detected in all three cell lines. In addition, UWOV2 (Sf) produces and secretes large amounts of extracellular matrix material
with an ordered fibrillar structure which may function as an attachment factor for the serum-free cells. These cell lines
seem to be useful for further studies of the biology of human ovarian cancer.
This research was supported by grants from National Cancer Association (S. A.) and Bekker Trust Foundation. 相似文献
12.
Sébastien Quesney Jacqueline Marvel Annie Marc Catherine Gerdil Bernard Meignier 《Cytotechnology》2001,35(2):115-125
The density of viable cells in a culture results from a balance between cell proliferation and cell death. The aim of this
study was to characterize and compare these two phenomena in Vero cell cultures in one serum containing medium (ScA) and one
serum free medium (SfB) in bioreactors. Cell growth was evaluated by cell counting(after crystal violet staining) and cell
cycle analysis. Necrosis and apoptosis were characterized and quantified by measuring the release of LDH, trypan blue exclusion,annex
in V-FITC/PI staining and TUNEL assay. ScA supported a higher maximal viable-cell density(2.3 × 106 vs. 1.8 × 106 cells ml-1). However, cell cycle analysis showed that cell division was more active in SfB than in ScA. LDH release in the supernatant
increased much earlier in SfB than in ScA (one vs. five days), but trypan blue counts showed no apparent difference in the
viability of the cultures. Apoptosis, evidenced by annexin V-FITC/PI staining, could be detected in the population of suspension
cells detached from microcarriers, but not among adherent cells; positivity of the TUNEL assay occurred later than that of
the annexin V-FITC/PI staining. Our data indicate that the lower cell yield in SfB,compared with that in ScA, results from
a higher cell death rate. Apparently, cells die from apoptosis followed by secondary necrosis.
This revised version was published online in July 2006 with corrections to the Cover Date. 相似文献
13.
Cultivation of mosquito cell lines in serum-free media and their effects on dengue virus replication
Goro Kuno 《In vitro cellular & developmental biology. Plant》1983,19(9):707-713
Summary Seven mosquito cell lines from five species (Aedes aegypti, Ae. albopictus, Ae. pseudoscutellaris, Culex tarsalis, andToxorhynchites amboinensis) were adapted to three kinds of serum-free media (SEM), which were composed of equal volumes of tryptose phosphate broth
and of either Leibovitz (L15) medium, Eagle’s minimum essential medium, or Medium 199 with Hanks’ salts. Population growth
rates of the cells cultivated in the SMFs were generally slower than those of original cell cultures maintained in conventional
media containing bovine sera. A karyological study showed a significant shift to heteroploidy in two of the four cell lines
examined. Four SMF-adapted sublines were compared with parental cultures for replication of dengue viruses.Ae. aegypti RML-12,Ae. albopictus C6/36,Ae. pseudoscutellaris AP-61, andTx. amboinensis TRA-171 demonstrated different levels of alteration in virus replication ranging from lower titers (as inAe. albopictus C6/36) to comparable or higher titers (as inAe. aegypti RML-12) when they were simultaneously inoculated with four dengue serotypes.
Use of trade names and commercial sources is for identification only and does not constitute endorsement by the Public Health
Service or by the U.S. Department of Health and Human Services. 相似文献
14.
M. Amouric J. Marvaldi J. Pichon F. Bellot C. Figarella 《In vitro cellular & developmental biology. Plant》1984,20(7):543-548
Summary Lactoferrin was examined for its effect on the growth of a human colon adenocarcinoma cell line (HT 29) in culture and its
action was compared to that produced by transferrin and two different iron solutions (ferrous sulfate and ferric chloride).
When transferrin was replaced by either iron solutions the cell grew in proportion to the quantity added and the maximal effect
obtained was identical to that produced by transferrin alone. When transferrin was replaced by lactoferrin the cells were
unable to proliferate for a long time. However, in the presence of low-concentration iron solutions, lactoferrin stimulated
the cell growth, and the effect was more pronounced with the ferric chloride solution.
This work was supported by Grants Inserum 817014 and LA CNRS 202. 相似文献
15.
A serum-free medium that supports the growth of cultured skeletal muscle satellite cells 总被引:2,自引:0,他引:2
Ronald E. Allen Michael V. Dodson Lynda S. Luiten Linda K. Boxhorn 《In vitro cellular & developmental biology. Plant》1985,21(11):636-640
Summary A serum-free medium has been devised that supports the proliferation and differentiation of primary cultures of rat skeletal
muscle satellite cells for up to 4 d. The medium consists of a mixture of Dulbecco's modified Eagle's medium and MCDB-104
plus insulin, dexamethasone, pituitary fibroblast growth factor, Deutsch fetuin, and linoleic acid. In addition to promoting
the formation of myotubes from satellite cells, a decrease in fibroblast contamination of these cultures was observed when
cultures grown in serum-free medium were compared to cultures grown in serum-containing medium.
This work was supported by the Arizona Agriculture Experiment Station, Project No. R11, U.S. Public Health Service Grant R01
AG03393, Lilly Research Laboratoires, and Merck Institute for Therapeutic Research. This communication is Arizona Agriculture
Experiment Station Journal Paper No. 3966. 相似文献
16.
Young N. Kim Young S. Park Hyun K. Kim Byung C. Jeon Se E. Youn Hyeon Y. Lee 《Cytotechnology》1993,13(3):221-226
The addition of ethanol extracts ofCentella asiatica showed a remarkable enhancement of fibroblast cells attachment to Cytodex beads in serum-free (SF) medium. It also improves tPA production in both batch and perfusion cultivations. The optimal concentration for SF medium was determined as 2 ppm of the extracts when using Cytodex III. In batch cultivation a high specific tPA production rate was obtained, compared to that from 5% FBS containing medium. However, a fast specific growth rate was observed in 5% FBS medium. In perfusion cultivation a reasonably good cell density and tPA production was achieved at a perfusion rate of 2.4×106 (viable cell/ml) and 0.65 (g/ml), respectively at 22 ml/min. 相似文献
17.
Development of a new serum-free medium,USC-HC1, for growth and normal phenotype in postembryonic chicken growth plate chondrocytes 总被引:1,自引:0,他引:1
Laura V. Hale John E. Hale Mary Lynn S. Kemick Yoshinori Ishikawa Roy E. Wuthier 《In vitro cellular & developmental biology. Plant》1986,22(10):597-603
Summary A serum-free medium for postembryonic chicken epiphyseal growth plate chondrocytes has been developed from 104 MCDB medium.
To enable these fastidious cells to survive, grow, and express normal phenotype, a substantial increase over MCDB 104 in the
level of many of the amino acids was required, as well as a change in the buffer system and the addition of SerXtend, a defined,
serum-free product containing various growth factors, including fibroblast growth factor. Also required was the provision
of cell attachment factors, either by coating culture surfaces with type II collagen, or better, by allowing the freshly released
cells to recover for several hours in a medium supplemented with 10% fetal bovine serum before plating. Ths new serum-free
medium, which we call USC-HC1, supports growth and replication, the retention of normal polygonal morphology, the expression
of significant levels of cellular alkaline phosphatase activity, the production of sulfated proteoglycans, type II collagen,
and the formation of alkaline phosphatase-rich matrix vesicles by the chondrocytes. The major advantage of USC-HC1, however,
is that it will provide for the first time an opportunity to examine the effects of various defined growth and hormonal factors
on the phenotypic expression and differentiation of growth plate chondrocytes, in the absence of the variable (stimulatory
and inhibitory) factors present in fetal bovine serum.
This work was supported by grant AM18983 from the National Institute of Arthritis, Diabetes, Digestive and Kidney Diseases,
Bethesda, MD. 相似文献
18.
Martine Chessebeuf Prudent Padieu 《In vitro cellular & developmental biology. Plant》1984,20(10):780-795
Summary Rat liver epithelial cells explanted in a serum-free medium (SFM) composed of Ham's F10 basal medium plus free fatty acids
adsorbed on bovine albumin gave successful rise to primary cultures and then to long-term cell lines that expressed liver
functions; induction ofl-tyrosine aminotransferase by glucocorticoids, hepatic pattern of progesterone metabolism, and biosynthesis of murine primary
bile acids; chenodeoxycholic and cholic acid common to higher vertebrates and α-muricholic acid specific of the rat bile. 相似文献
19.
We tried to establish a new serum-free and heat-sterilizable medium, based on our serum-free medium in which many lymphoblastoid cells and hybridoma could grow as well as in a conventional serum-containing medium.As is well-known, L-glutamine (L-Gln) is one of the most heat-labile but essential components for cell growth. As a substitute for L-Gln, dipeptide such as Gly-L-Gln or L-Ala-L-Gln, which was quite stable even after autoclaving, was found to be utilizable for mammalian cell growth. The L-Gln dipeptide-containing serum-free medium was quite stable in a solution even after storing at 37°C for 4 months. In the serum-free medium containing L-Ala-L-Gln, mouse hybridola could grow and produce more antibody than in RPMI 1640+10% FBS. 相似文献
20.
Yoshiko Myoken Yoshinari Myoken Tetsuji Okamoto Mikio Kan J. Denry Sato Kazuaki Takada 《In vitro cellular & developmental biology. Animal》1994,30(11):790-795
Summary A squamous cell carcinoma cell line Nakata proliferated in serum-free culture and was not responsive to exogenous fibroblast
growth factor-1 (FGF-1). Immunostaining revealed that Nakata cells expressed FGF-1 in their cytoplasms and nuclei. Two molecular
mass species of FGF-1 (16 and 18 kDa) were identified in cell extracts by Western blot. These cells also expressed high-affinity
FGF-1 binding sites (Kd=360 pM, 28 000 sites/cell). The results of cross-linking with [125I]FGF-1 demonstrated the presence of two bands with molecular masses of 160 and 140 kDa. The addition of FGF-1 specific antisense
oligonucleotides at 25 μM to Nakata cells resulted in an 82% inhibition in cell growth and suppressed FGF-1 expression. This effect was dose-dependent
and specific, because sense oligonucleotides were ineffective in inhibiting cell growth. In addition, Nakata cell growth was
suppressed by an anti-FGF-1 neutralizing antibody, which resulted in a 52% inhibition at 8 μg/ml. These results demonstrate
that Nakata cells produce FGF-1, and indicate that this growth factor acts in an autocrine manner by interacting with FGF-1
binding sites on Nakata cells. 相似文献