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1.
A saline extract was prepared from Drosophila eggs. When diluted to a concentration of 1% with Drosophila tissue culture medium, it did not support growth of cells from the Drosophila line D1 during the first few days of subculture as well as medium containing serum. When cells reached a stationary phase, however, the cell density in medium containing extract was greater than in medium containing serum. By altering the concentrations of the extract, and by adding bovine albumin, a medium was obtained in which D1 cells survived initial culturing, and which supported cell growth by day 4 as well as medium plus serum. The initial retardation of growth in medium containing egg extract might be due to the need of the cells to adapt to the new medium. At the present time four Drosophila cell lines have been maintained in this medium for more than 16 passages. Preliminary experiments with primary embryonic Drosophila cells indicate that medium containing 2% extract and bovine albumin retards the differentiation of these cells. 相似文献
2.
Kedar N. Prasad Erika Carvalho Judith Edwards-Prasad Francisco G. La Rosa Sanjay Kumar Jae Ho Kim Arlen Meyers Susan Kentroti 《In vitro cellular & developmental biology. Animal》1994,30(5):312-320
Summary This study reports the establishment ofα-amylase-producing human parotid pleomorphic adenoma cell lines (2HP and 2HP1) which have been maintained in culture for over 1 yr. The procedures required preparation of cellular clumps from tumor tissue
and plating them on plasma clot or precoated dishes. During the initial phase of growth they required modified MCDB-153 medium
without serum. When cells showed signs of degeneration they were changed to MCDB-153 medium containing first 2% and then 10%
heat inactivated fetal bovine serum. Although cells grew well in MCDB-153 containing 10% serum, the epithelial cell morphology
was not distinct. Therefore, the growth and morphology of cells grown in MCDB-10% serum were compared with those in RPMI growth
medium containing 10% fetal bovine serum and F12 containing 10% agammaglobulin newborn bovine serum. Although the growth of
cells was a little slower in F12 medium than those in MCDB and RPMI, the epithelial cell morphology was maintained better
than in other growth media. The cells of 2HP and 2HP1 produce low levels ofα-amylase and relatively high levels ofα-amylase mRNAs of 1176 and 702 bp and contain neurofilament-160, a neuronal-specific marker. The cells of 2HP1 are tumorigenic when tested in athymic mice, but the cells of 2HP are not. The establishment of amylase-producing human parotid
adenoma cell lines of different characteristics in culture provides a new opportunity to study the mechanisms of differentiation
and transformation, and regulation ofα-amylase in these cells. 相似文献
3.
I Yamane O Murakami M Kato 《Proceedings of the Society for Experimental Biology and Medicine. Society for Experimental Biology and Medicine (New York, N.Y.)》1975,149(2):439-442
A serum-free culture medium, supplemented with 1% bovine serum albumin, supported the growth of both primary and continuous suspension-type cultures of various mammalian tumor cells. The role of albumin added to the medium was also studied. Defatted albumin failed to support cell growth, unless reconstituted with its lipid extract. Similarly, defatted albumin when combined with oleic and linoleic acids, also supported cell growth. Therefore, albumin-bound fatty acids play an important growth-promoting role in serum-free medium. 相似文献
4.
The glutamine requirement for thein vitro proliferation of fish cells was investigated with cell lines from four different species and three tissues: goldfish skin (GFSk-S1), Chinook salmon embryo (CHSE-214), and raibow trout liver (RTL-W1) and spleen (RTSp-W1). With a supplement of fetal bovine serum, the basal medium, Leibovitz's L-15, without glutamine supported the proliferation of all four cell lines as well, or nearly as well, as L-15 with 2 mM glutamine. This was true over short term assays of two to four weeks and for continuous propagation. CHSE-214 also grew as well with or without 2 mM glutamine in Minimum Essential Medium with fetal bovine serum. However, when the supplement was dialyzed fetal bovine serum, CHSE-214 grew much better in L-15 without glutamine. Therefore, glutamine was not required for growth in L-15, and in fact, was inhibitory in the absence of the dialyzable fraction of serum. By contrast, glutamine appeared to be important for growth in Minimum Essential Medium. When the supplement was dialyzed fetal bovine serum, CHSE-214 grew much better in Minimum Essential Medium with 2 mM glutamine. These results suggest that the glutamine requirement for thein vitro proliferation of fish cells is conditional and depends on the basal medium and serum supplement.Abbreviations BSA
bovine serum albumin
- CHSE-214
Chinook samon embryo cell line
- dFBS
dialyzed fetal bovine serum
- FBS
fetal bovine serum
- GFSk-S1
goldfish skin cell line
- GS
glutamine synthetase
- L-15
Leibovitz's L-15 media
- L929
mouse fibroblast cell line
- MEM
minimum essential medium Eagle
- PBS
phosphate buffered saline
- RTL-W1
rainbow trout liver cell line
- RTSp-W1
rainbow trout spleen cell line 相似文献
5.
Merrill S. Babcock Maria R. Marino William T. Gunning III Gary D. Stoner 《In vitro cellular & developmental biology. Plant》1983,19(5):403-415
Summary The clonal growth and serial propagation of rat esophageal epithelial cells in low serum-containing medium has been achieved
without feeder layers or conditioned medium. To date, a total of four lines have been developed and maintained for as many
as 40 passages in culture. Growth of the cells was possible only after modifying the culture medium (PFMR-4) by reducing the
calcium concentration from 1 to 0.1 mM, and by adding low levels of dialyzed fetal bovine serum and seven growth factors; i.e. epidermal growth factor, hydrocortisone,
ethanolamine, phosphoethanolamine, insulin, transferrin, and cholera toxin. Cell lines have been developed from both explant
outgrowths and enzyme dissociated esophagi. The epithelial nature of the cells was confirmed by electron microscopy and immunological
methods. Clonal growth studies revealed that optimal cell growth occurred in medium containing 2.4% dialyzed fetal bovine
serum and 0.1 mM calcium. Calcium levels of 0.3 mM or higher caused the cells to stratify and undergo terminal differentiation. Coating the culture dishes with collagen, or
a combination of collagen, fibronectin, and bovine serum albumin, increased both the cell growth rate and the colony forming
efficiency. The successful long term culture of rat esophageal epithelial cells permits their use as models in studies concerned
with esophageal differentiation and carcinogenesis.
This investigation was supported by U.S. Public Health Service Grant CA 28950, awarded by the National Cancer Institute, Bethesda,
MD. 相似文献
6.
Growth of mouse vaginal epithelial cells in culture: Effect of sera and supplemented serum-free media 总被引:3,自引:0,他引:3
Taisen Iguchi Francis-Dean A. Uchima Howard A. Bern 《In vitro cellular & developmental biology. Plant》1987,23(8):535-540
Summary Normal mouse vaginal epithelial cells isolated from ovariectomized ca. 35-d-old BALB/cCrgl mice were grown in primary culture
using collagen gel metrix and a serum-free medium composed of a 1∶1 mixture of Dulbecco’s modified Eagle’s medium and Ham’s
F12 (D:H) medium supplemented with insulin (IN), epidermal growth factor (EGF), cholera toxin, transferrin, and bovine serum
albumin V (BSA). Three-dimensional cellular outgrowths occurred inside the collagen gel matrix. The contribution of each factor
to cell growth was examined by individual addition to the basic D:H medium and by individual deletion from the complete serum-free
medium. When added individually, only IN promoted growth. Deletion of IN from the complete serum-free medium markedly, diminished
growth; deletion of EGF or BSA slightly diminished growth. When horse, fetal bovine, or chicken serum was added to the basal
D:H medium, only with increasing doses of horse serum was there enhanced cell growth. The effect of 17?-estradiol and diethylstilbestrol
on the growth of cells was also tested, using a suboptimal medium of D:H supplemented with BSA and IN, or a minimal medium
supplemented with IN alone. During the 8-d time period, addition of estrogen did not enhance cell growth in either medium.
To date, we have been unable to demonstrate a mitogenic effect of estrogen; rather a dose-dependent inhibition of proliferation
is seen.
This investigation was supported by grants CA-05388 and CA-09041, awarded by the National Institutes of Health, DHHS, Bethesda,
MD. 相似文献
7.
Gary L Bradshaw Gordon H. Sato Don B. McClure George R. Dubes 《Journal of cellular physiology》1983,114(2):215-221
A serum-free medium for serial culture of baby hamster kidney cell line 21 (BHK-21) as container-adherent cells was developed. The medium is a 1:1 (v/v) mixture of Dulbecco's modified Eagle's medium and Ham's F-12 medium supplemented with fibroblast growth factor, fibronectin, insulin, oleic acid (preincubated with fatty-acid-free bovine serum albumin as carrier), and transferrin. The fibronectin was required for cell adherence, the other factors for optimal cell multiplication. When cell input was greater than about 1,900 cells/cm2, this serum-free medium supported cell multiplication at a rate approximately equal to the rate in medium with 10% serum. At lower cell input, growth in the serum-free medium was poor unless it was supplemented with serum-free medium which had been conditioned by BHK-21 cells. The conditioned medium contained a factor(s) which enabled or stimulated cell multiplication. 相似文献
8.
Growth of MTW9/PL2 estrogen-responsive rat mammary tumor cells in hormonally defined serum-free media 总被引:3,自引:0,他引:3
David Danielpour Terry L. Riss Masami Ogasawara David A. Sirbasku 《In vitro cellular & developmental biology. Plant》1988,24(1):42-52
Summary Growth of the estrogen-responsive MTW9/PL2 rat mammary tumor cells was demonstrated in serum-free defined medium (designated
DDM-1) formulated with F12-DME (1:1 vol/vol) supplemented with 15 mM HEPES pH 7.4 insulin 10 μg/ml, transferrin 10 μg/ml, sodium selenite 10 ng/ml, triiodo-l-thyronine 0.3 nM, phosphoethanolamine 5 μM, epidermal growth factor (20 ng/ml), 17 β-estradiol 2 nM, and bovine serum albumin 20 μg/ml. In DDM-1, the growth rate was about one-half that seen in serum-containing medium. When
ethanolamine (50 μM), glutathione (20 μg/ml), and linoleic acid/bovine serum albumin (150 μg/ml) were added (formulation DDM-2), the growth rate
was 80% of serum-containing medium and not seed-density dependent. Deletion of estradiol from DDM-1 or DDM-2 had no effect
on growth rate. Also, cells grown in steroid hormone deficient medium for 4 mo. continued to form estrogen-responsive tumors
in rats as did cells cultured for the same period in 2 nM estradiol. To investigate autocrine growth factor secretion, a third medium (DDM-3) was prepared by deleting insulin, epidermal
growth factor, phosphoethanolamine, estradiol, and both forms of bovine serum albumin from DDM-2. Growth in mitogen-free medium
equaled 86% of the serum-stimulated rate and was seed-density dependent; phenol red deletion from DDM-3 had no effect on growth
rate. Evidence presented suggests that autocrine factors stimulate growth of the MTW9/PL2 cells in DDM-3, and that this secretion
may support the growth of estrogen-responsive cells in culture in the absence of steroid hormone.
This work was supported by National Cancer Institute grants CA-38024 and CA-26617 and American Cancer Society grant BC255. 相似文献
9.
David G. Thomassen 《In vitro cellular & developmental biology. Animal》1993,29(6):498-504
Summary Factors regulating the proliferation of normal, preneoplastic, and neoplastic rat tracheal epithelial (RTE) cells were investigated
to identify changes taking place during the progression of RTE cells to neoplasia. Normal RTE cells exhibit clonal proliferation
in a serum-free medium containing pituitary extract, serum albumin, cholera toxin, epidermal growth factor, hydrocortisone,
and insulin. All combinations of these six factors were examined for their abilities to support clonal proliferation of normal,
preneoplastic, and neoplastic RTE cells. In general, preneoplastic RTE cells required fewer factors for proliferation than
normal RTE cells, and neoplastic cells required fewer factors than preneoplastic cells. A common pattern of reductions has
been identified in the growth factors required for the clonal proliferation of preneoplastic vs. normal RTE cells and for
neoplastic vs. preneoplastic and normal RTE cells. Normal RTE cells exhibit clonal proliferation in a serum-free medium supplemented
with a minimum of six factors: bovine serum albumin, bovine pituitary extract, cholera toxin, epidermal growth factor, hydrocortisone,
and insulin. Preneoplastic RTE cells exhibit clonal proliferation in a serum-free medium supplemented with four factors: bovine
serum albumin, bovine pituitary extract, hydrocortisone, and insulin. Finally, neoplastic RTE cells exhibit clonal proliferation
in a serum-free medium supplemented with two factors: bovine serum albumin and bovine pituitary extract. These results suggest
that the progression of RTE cells to neoplasia is associated with a series of changes in regulatory pathways that control
cell proliferation. 相似文献
10.
Summary L cells were grown in spinner cultures in a defined medium consisting of Waymouth medium MB752/1 (19) supplemented with 2
mg of fatty acid-free bovine serum albumin (BSA) per ml and 5 μg of oleate per ml (WO5 medium). Growth in WO5 medium was comparable to spinner L cell growth in two serum-containing media. The optimal concentration of oleate in the
WO medium was 5 to 10 μg per ml. The use of 20 to 80 μg of oleate per ml of medium resulted in lower peak populations and
earlier declines in viable cell counts. Cell death occurred rapidly in WO160 medium. Cell growth in WO medium containing 5 to 80 μg of oleate per ml was well above the level of growth observed when
no oleate was present in the medium. Since the total lipid and fatty acid compositions of the BSA used in this study have
been characterized by the authors, the WO medium may be considered a defined medium. L cells have been continuously maintained
in spinner cultures in WO5 medium for over 50 passages with no major variation in the growth pattern. A 1000-fold increase inChlamydia psittaci strain meningopneumonitis, with a peak titer of 9.3×107 plaque-forming units per ml, was observed when the chlamydial agents were grown in spinner L cells in WO5 medium.
This investigation was supported by Public Health Service Research Grant HE 08214 from the Program Projects Branch, Extramural
Programs, National Heart and Lung Institute; The World Health Organization; and The Hormel Foundation. 相似文献
11.
Improved media for normal human muscle satellite cells: Serum-free clonal growth and enhanced growth with low serum 总被引:5,自引:0,他引:5
Richard G. Ham Judy A. St. Clair Cecelia Webster Helen M. Blau 《In vitro cellular & developmental biology. Plant》1988,24(8):833-844
Summary We have developed a serum-free medium for clonal growth of normal human muscle satellite cells (HMSC). It consists of an optimized
nutrient medium MCDB 120, plus a serum-free supplement, designated SF, that contains epidermal growth factor (EGF), insulin,
dexamethasone, bovine serum albumin, and fetuin. Fibroblast growth factor was needed with dialyzed fetal bovine serum (dFBS)
as the only other supplement, but in media containing SF, it was only slightly beneficial, and was omitted from the final
medium without significant loss. Clonal growth of HMSC in MCDB 120 plus SF is as good as with 15% serum and 0.5% chicken embryo
or bovine pituitary extract. However, growth is further improved by use of a doubly-supplemented (DS) medium containing both
SF and 5% dFBS. Clonal growth of HMSC in the DS medium far exceeds that in previous media with any amount of serum, and monolayer
growth is at least equal to that in conventional media with higher levels of serum. Cells grown in these media exhibit little
differentiation, even when grown to high densities. However, they retain the capacity for extensive fusion and synthesis of
increased creatine kinase when transferred to a serum-free differentiation-promoting medium, such as Dulbecco's modified Eagle's
medium plus insulin. All experiments were done with clonal cultures of HMSC to insure that observed growth responses were
always those of muscle cells.
This research was supported by a grant from the Muscular, Dystrophy Association.
Editor's statement This article describes the optimization of both the basal nutrient medium and growth factor requirements
for human muscle cells in vitro. This system is critical for studies of normal muscle cell and molecular biology, as well
as for understanding diseases of muscle such as Duchenne, Muscular Dystrophy. 相似文献
12.
Atsuyuki Okuda Yoshitsugu Kajiwara Genki Kimura 《In vitro cellular & developmental biology. Plant》1983,19(5):376-384
Summary A semiserum-free medium was developed for monolayer culture of rat 3Y1 fibroblastic cells. The main components of the developed
medium added to Dulbecco's modified Eagle's medium (DMEM) were insulin transferrin epidermal growth factor, poly-d-lysine, bovine albumin, oleic acid, and bovine α-globulin. In this medium, 3Y1 cells grew in mass culture at much the same
rate as in DMEM supplemented with 10% fetat bovine serum (FBS), and colonies, albeit of smaller sizes, diddform. Virally transformed
derivatives of 3Y1 (simian virus 40-3Y1, polyoma virus-3Y1 and adenovirus type 12-3Y1) also formed colonies in the semiserum-free
medium.
When trypsinized 3Y1 cells were seeded with the medium lacking α-globulin, neither growth in the mass culture nor clonal growth
in the low density culture (clonal growth) occurred. In this case, cell spreading was inhibited by albumin, and this inhibition
was overcome by adding α-globulin or treating dishes with serum. When albumin was excluded from the semiserum-free medium,
clonal growth did not occur, whereas growth in mass culture and stimulation of DNA synthesis in the resting mass culture (stimulation
of DNA synthesis) were not so drastically affected. When oleic acid was removed, growth in mass culture was inhibited considerably,
but no considerable effect was seen on clonal growth or on stimulation of DNA synthesis. In the absence of insulin, stimulation
of DNA synthesis was inhibited more markedly than when other components were removed, but such was not the case with growth
in mass culture and clonal growth. 相似文献
13.
J. H. Wharton R. D. Ellender B. L. Middlebrooks P. K. Stocks Adrian R. Lawler D. Howse 《In vitro cellular & developmental biology. Plant》1977,13(6):389-397
Summary A cell line designated SP-1 was established from tissue of the silver perch,Bairdiella chrysura. Cells were fibroblast-like and grew best at 26°C in Leibovitz medium (L-15) containing 15% fetal bovine serum and 0.150m sodium chloride. Passage 1 to passage 9 SP-1 cells contained a chromosome number of 48; at passages 27 and 50 the modal numbers
were 51 and 54, respectively. Confirmation of the origin of SP-1 cells was made by the cytotoxic antibody dye-exclusion test.
This cell line supported the growth of lymphocystis virus from the silver perch but was not found to replicate various other
fish and mammalian viruses.
This study was supported in part by fund supplied by the Faculty Research Council of the University of Southern Mississippi
and by the National Oceanographic and Atmospheric Administration, Office of Sea Grant No. 04-3-158-58. A portion of these
results were presented at the 26th Annual Meeting of the Tissue Culture Association, Motreal, Canada, 1975. 相似文献
14.
NS0 has been used as a fusion partner for the production of hybridomas and has more recently been engineered to produce recombinant protein. A protein-free culture medium, designated W38 medium, has previously been developed which supported high density growth of rat myeloma and hybridoma cell lines. NS0 cells failed to grow in W38 medium and in a number of protein-free culture media which support the growth of other myeloma cell lines. NS0 cells are derived from the NS-1 cell line, which is known to require exogencus cholesterol. It was found that NS0 cells grew in W38 medium supplemented with phosphatidylcholine, cholesterol, and albumin and that NS0 were auxotrophic for cholesterol. Protein-free growth of NS0 cells was achieved by using -cyclodextrin to replace albumin as a lipid carrier. The maximal cell density reached in this protein-free medium was in excess of 1.5×106 cell ml–1. The lipid supplements in the medium precipitated after a few days storage at +4°C. In order to overcome this problem a protocol was developed which allowed NS0 cells to be adapted to cholesterol-independent growth in W38 medium. NS0.CF (cholesterol-independent NS0 cells) were cultured continuously in W38 medium for several months. In shake flask culture a cell density of 2.4×106 cells ml–1 was achieved in W38 medium compared with 1.41×106 cells ml–1 in RPMI 1640 medium containing 10% foetal bovine serum. NS0.CF cells readily grew in a 1 litre stirred bioreactor using W38 medium supplemented with Pluronic F68 reaching a density of 3.24×106 cells ml–1. NS0.CF were cloned protein-free by limiting dilution in W38 medium, giving colonies in wells that were seeded at an average density of 0.32 cells per 200 l. This study has demonstrated for the first time the growth of a cholesterol-requiring mouse myeloma cell line in a completely defined protein-free medium and its subsequent adaptation to cholesterol-independence.Abbreviations BSA
bovine serum albumin
- C
cholesterol
- CD
cyclodextrin
- F68
Pluronic F68
- GS
glutamine synthetase
- P
phosphatidylcholine
- PC-FBS
phosphatidylcholine, cholesterol and foetal bovine serum
- RPMI
RPMI 1640 medium
- MSX
methionine sulphoximine 相似文献
15.
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
16.
Akio Nishikawa Keiko Shimizu-Nishikawa Leo Miller 《In vitro cellular & developmental biology. Plant》1990,26(12):1128-1134
Summary Methods for the isolation and in vitro culture of larval and adultXenopus laevis epidermal cells have been developed. Epidermal cells of stage 52–54 tadpoles and adult epidermal cells were enzymatically
dissociated and purified (98%) by Percoll-density centrifugation and unit-gravity sedimentation. Both cell types attached
on fibronectin-coated dishes and proliferated for 1 wk when the proper medium was used. There were four significant differences
between larval and adult cells: a) Adult cells had a greater buoyant density than larval cells. b) Keratin synthesis patterns
were markedly different. c) A combination of medium F12 and Eagle's minimum essential medium was optimal for growth of larval
cells whereas MCDB151 medium was optimal for adult cells. d) Adult cells needed fetal bovine serum (>5%) whereas larval cells
grew without fetal bovine serum. In contrast to these differences, larval and adult cells had two similar properties: a) Insulin
had a potent effect on the growth of both cells, and b) The optimal Ca++ concentration for cell growth was quite low for both cell types; 0,1 mM for larval cells and below 0.05 mM for adult cells. These results suggest that low Ca++ levels are essential for both cornifying (adult) and uncornifying (larval) amphibian keratinocytes. The culture techniques
described herein for larval and adult epidermal cells provide a new in vitro model for analyzing development of the epidermis
during amphibian metamorphosis.
This study was supported by grant (HD 24438) from the National Institutes of Health, Bethesda, MD. 相似文献
17.
Eun-Ho Park Jae-Seong Lee Ae-Kyng Yi Hisami Etoh 《In vitro cellular & developmental biology. Plant》1989,25(11):987-994
Summary A cell line (ULF-23HU) from the fin of the central mudminnow (Umbra limi) was characterized and tested for its suitability to assess cytogenetic damages induced by chemicals in fish. Cells of this
line exhibit a fibroblastlike appearance and grew optimal at 25°C in, TC-199 medium containing 10% fetal bovine serum, but
slower growth continued down to 4°C, where they could be stored for prolonged periods. Seeding efficiency of ULF-23HU cells
on the plastic substratum was approximately 85% in the above culture medium at 25°C. They had a 32-h cell cycle time taken
up by a 20-h S period as determined by the autoradiographic analysis of the fraction of labeled mitosis. Cultures showed relatively
high mitotic index (0.84 to 2.35%) during exponential growth phase lasting about 7 d. Karyological analysis of the cells at
the different subculture passages revealed constant chromosome modal number of 23 consisting of metacentric or submetacentric
chromosomes, which were primarily similar to those of in vivo cells, with one additional chromosome. The spontaneous sister
chromatid exchange rate was 5.3 per metaphse. When ULF-23HU cells were exposed toN-nitroso-N-methylurea, a clastogen in the mammalian cells, dose-dependent increases both in sister chromatid exchanges and chromosome
aberrations were clearly detected. These results on the growth kinetics and cytogenetic characteristics offered the high possibility
of the use of this cell line as a suitable in vitro model for clastogenicity studies in fish.
This work was supported by grants from Korea Sciences and Engineering Foundation and Japanese Government Research Awards for
Foreign Specialists to E.-H. Park. 相似文献
18.
Peter M. Gresshoff 《Archives of microbiology》1981,128(3):303-306
Chlamydomonas reinhardi can utilise the lower aliphatic amides (C1–C4) as nitrogen sources. Of these only acetamide can serve as a sole carbon source. The acetamide analogue F-acetamide kills cells after conversion to F-acetate and F-citrate. This conversion is controlled by exogenous ammonia and, in part, acetate levels. Only one enzyme and one active site are involved in acetamidase function. Enzymatic analysis indicates an increased substrate range as compared to the growth — supported range, indicating uptake, toxicity or metabolic control restrictions.Abbreviations TCA
trichloroacetic acid
- TAP
tris-acetate-phosphate medium
- MIC
mimmum inhibitory concentration
- BSA
bovine serum albumin 相似文献
19.
In this work, water-soluble extracts of Ganoderma lucidum spores (Gls), a Chinese medicinal herb that possesses cell growth stimulating function, were found to be an effective growth factor for Chinese hamster ovary (CHO) cell cultivation. The Gls extract was prepared and supplemented to CHO K1 cell culture media with various serum levels. Our results obtained from both the static culture and the spinner-flask suspension culture showed that use of small-amount Gls extract effectively promoted cell growth and suppressed cell apoptosis induced by serum deprivation with normal cell cycle maintained in a low-serum medium. The low-serum medium containing 1 % (v/v) fetal bovine serum (FBS) and 0.01 % (w/v) Gls extract showed a comparable performance on both cell growth and fusion protein productivity with the conventional CHO culture medium containing 10 % (v/v) FBS and a commercial serum-free medium. This is the first study of the potential of Gls extracts for use as an alternative cell growth factor and nutrient for CHO cells. The findings have presented a new approach to economic cultivation of CHO cells for therapeutic protein production. 相似文献
20.
ZF-L cells were derived from normal adult zebrafish liver, and have been growing in culture for more than 100 generations. The cells were derived in basal nutrient medium supplemented with fetal bovine serum (FBS), trout serum, trout embryo extract, bovine insulin and mouse epidermal growth factor. After 50 generations in culture, optimal growth of the cells was achieved in medium supplemented with FBS (5%) and trout serum (0.5%). ZF-L cells were hypodiploid (modal chromosome number= 46) and exhibited an epithelial morphology. ZF-L cell homogenates exhibited alanine and aspartate aminotransferase, glucose-6-phosphatase and alkaline phosphatase enzyme activities. The cells synthesized and released several proteins into the culture medium, including a 70 kDa protein recognized by anti-bovine serum albumin IgG.Abbreviations NF
-naphthoflavone
- EGF
epidermal growth factor
- EROD
7-ethoxyresorufinO-deethylase
- FBS
fetal bovine serum
- PBS
phosphate-buffered saline
- PMSF
phenylmethylsulfonyl fluoride
- TCDD
2,3,7,8-tetrachlorodibenzo-p-dioxin 相似文献