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1.
Donna M. Peehl Stephen T. Wong Thomas A. Stamey 《In vitro cellular & developmental biology. Plant》1988,24(6):530-536
Summary The ability of human epithelial cells derived from adult prostatic tissues to undergo clonal growth in culture was examined.
In a previously described serum-free culture system, such cells exhibited a density-dependent growth requirement. It was found
that raising the level of one of the constituents of the culture medium, bovine pituitary extract, to 100 μg/ml permitted
excellent clonal growth when as few as 100 cells were inoculated/60-mm2 dish. Raising the levels of supplements other than pituitary extract (cholera toxin, epidermal growth factor, hydrocortisone,
or insulin) did not produce this result. The average colony-forming efficiency of cells derived from primary or early passage
cultures was approximately 25%. When single cell suspensions were prepared from tissue isolates and directly analyzed for
clonal growth, colony-forming efficiencies were approximately 5%, perhaps indicating the proportion of stem cells with proliferative
potential in the original isolates. The colony-forming efficiency of a cell population derived from cancer tissue was not
significantly different from those of populations derived from normal tissues. 相似文献
2.
Olfactory neuroepithelium (ONe) is unique because it contains progenitor cells capable of mitotic division that replace damaged or lost neurons throughout life. We isolated populations of ONe progenitors from adult cadavers and patients undergoing nasal sinus surgery that were heterogeneous and consisted of neuronal and glial progenitors. Progenitor lines have been obtained from these cultures that continue to divide and form nestin positive neurospheres. In the present study, we used clonal and population analyses to probe the self-renewal and multipotency of the neurosphere forming cells (NSFCs). NSFCs plated at the single cell level produced additional neurospheres; dissociation of these spheres resulted in mitotically active cells that continued to divide and produce spheres as long as they were subcultured. The mitotic activity of clonal NSFCs was assessed using bromodeoxyuridine (BrdU) incorporation. Lineage restriction of the clonal cultures was determined using a variety of antibodies that were characteristic of different levels of neuronal commitment: β-tubulin isotype III, neural cell adhesion molecule (NCAM) and microtubule associated protein (MAP2), or glial restriction: astrocytes, glial fibrillary acidic protein (GFAP); and oligodendrocytes, galactocerebroside (GalC). Furthermore, nestin expression, a marker indicative of progenitor nature, decreased in defined medium compared to serum-containing medium. Therefore, adult human ONe-derived neural progenitors retain their capacity for self-renewal, can be clonally expanded, and offer multipotent lineage restriction. Therefore, they are a unique source of progenitors for future cell replacement strategies in the treatment of neurotrauma and neurodegenerative diseases. 相似文献
3.
Olfactory neuroepithelium (ONe) is unique because it contains progenitor cells capable of mitotic division that replace damaged or lost neurons throughout life. We isolated populations of ONe progenitors from adult cadavers and patients undergoing nasal sinus surgery that were heterogeneous and consisted of neuronal and glial progenitors. Progenitor lines have been obtained from these cultures that continue to divide and form nestin positive neurospheres. In the present study, we used clonal and population analyses to probe the self-renewal and multipotency of the neurosphere forming cells (NSFCs). NSFCs plated at the single cell level produced additional neurospheres; dissociation of these spheres resulted in mitotically active cells that continued to divide and produce spheres as long as they were subcultured. The mitotic activity of clonal NSFCs was assessed using bromodeoxyuridine (BrdU) incorporation. Lineage restriction of the clonal cultures was determined using a variety of antibodies that were characteristic of different levels of neuronal commitment: β-tubulin isotype III, neural cell adhesion molecule (NCAM) and microtubule associated protein (MAP2), or glial restriction: astrocytes, glial fibrillary acidic protein (GFAP); and oligodendrocytes, galactocerebroside (GalC). Furthermore, nestin expression, a marker indicative of progenitor nature, decreased in defined medium compared to serum-containing medium. Therefore, adult human ONe-derived neural progenitors retain their capacity for self-renewal, can be clonally expanded, and offer multipotent lineage restriction. Therefore, they are a unique source of progenitors for future cell replacement strategies in the treatment of neurotrauma and neurodegenerative diseases. 相似文献
4.
Dr. Donna M. Peehl Richard G. Ham 《In vitro cellular & developmental biology. Plant》1980,16(6):526-540
Summary A survey of commercially availabla media revealed that medium F-12 was superior to medium 199 for clonal growth of human epidermal
keratinocytes (HK) when supplemented with 10 μg/ml hydrocortisone (HC) plus dialyzed fetal bovine serum protein (FBSP), rather
than the whole serum used in previous studies. Qualitative and quantitative adjustment of the medium composition for optimal
clonal growth with minimal amounts of FBSP generated a new medium, MCDB 151, which supports clonal growth of HK with 10 μg/ml
HC and as little as 1 mg/ml FBSP (equivalent in protein concentration to 2.0% whole serum). MCDB 151 differs significantly
from MCDB 105, previously developed in this laboratory for normal human fibroblasts, and each medium selectively favors growth
of its own type of cell in primary cultures of disaggregated human neonatal foreskin cells. Differences in the amounts of
calcium and adenine in the two media appear to be among the most influential factors mediating the selective growth. Optimal
growth of HK occurs at a very low level of Ca2+ that causes the colonies to remain as monolayers rather than stratifying as they do in the presence of higher levels of calcium.
However, keratin synthesis, which was examined through use of highly specific fluorescent antibodies, is not affected by the
Ca2+ concentration. Agents that increase intracellular cyclic AMP levels appear to have no effect on HK growth in MCDB 151 with
10 μg/ml HC and 1.0 mg/ml FBSP.
This paper contains material from a thesis submitted to the Graduate School of the University of Colorado, Boulder, by Donna
M. Peehl in partial fulfillment of the requirements for the Ph.D. degree.
This work was supported by Grant CA 15305 from the National Cancer Institute and Grant AG 00310 from the National Institute
on Aging. 相似文献
5.
Summary The tumor promoter phorbol 12,13-didecanoate (PDD) significantly altered the growth properties of early passage normal human
skin cells in vitro in culture medium supplemented with elevated concentrations of selected amino acids. Continuous treatment
of cell with 10−7 or 10−8
M PDD resulted in a 5 to 10-fold increase in saturation density at early passages followed by a long-term two- to fourfold
increase. The PDD-treated cultures remained in exponential growth at cell densities greater than 10-fold higher than the control
cultures. Removal of PDD from the culture medium while the cells were at a high cell density resulted in a return to near-normal
saturation density by the subsequent passage. Anchorage independent growth of normal human cells in methylcellulose was also
promoted by PDD in a dose dependent manner, with prior subculturing in the presence of PDD being required for maximal colony
formation. The structural analog 4α-phorbol 12,13-didecanoate failed to elicit similar cellular responses.
This work was supported by grants from the Air Force Office of Scientific Research, Department of Defense (AFOSR-80-0283)
and the National Cancer Institute, Department of Health and Human Services (P-30-CA-16058). 相似文献
6.
Development of improved media and culture conditions for clonal growth of normal diploid cells 总被引:1,自引:0,他引:1
Richard G. Ham Wallace L. McKeehan 《In vitro cellular & developmental biology. Plant》1978,14(1):11-22
Summary Multiplication of normal diploid cells in culture is controlled by a complex set of interacting extracellular variables. The
amount of serum protein needed for colony formation by such cells is affected directly by many of the other variables, including
the nature of the culture surface, the type of trypsinization procedure used, and the qualitative and quantitative composition
of the culture medium. By a sequential process of adjusting all of these variables to optimum values for cellular multiplication
with minimal amounts of serum protein, we have been able to obtain clonal growth of normal human and chicken cells with less
than 500 μg per ml dialyzed serum protein. Precise quantitative adjustment of nutrient concentrations is particularly important.
The multiplication-promoting functions of serum can be classified operationally as “replaceable” (those that can be replaced
by modifying the medium or the culture conditions) and “nonreplaceable” (those that we have not yet been able to replace).
Elimination of the requirement for replaceable functions of serum has improved greatly the specificity and sensitivity of
the bioassay for the nonreplaceable functions. The nonreplaceable multiplication-promoting activity from fetal bovine serum
for human diploid fibroblasts has been separated from fetuin and serum albumin and purified approximately 15-fold.
Presented in the Opening Symposium on Nutritional Factors and Differentiation at the 28th Annual Meeting of the Tissue Culture
Association, New Orleans, Louisiana, June 6–9, 1977.
This work was supported by Contract 223-74-1156 from the Bureau of Biologics, U.S. Food and Drug Adminsstration, Grant AG
00310 from the National Institute on Aging, and Grant CA 15305 from the National Cancer Institute. 相似文献
7.
W. Gregory Hamilton Richard G. Ham 《In vitro cellular & developmental biology. Plant》1977,13(9):537-547
Summary A protein-free synthetic medium, MCDB 301, has been developed for clonal growth of Chinese hamster ovary cell lines. Medium
F12 was developed originally for that purpose, but later failed to support good growth without small amounts of serum protein.
Growth was restored by the addition of nonphysiological amounts of commercially prepared thyroxine or smaller amounts of the
trace element selenium. The thyroxine preparation was shown to contain sufficient selenium to account for all of its growth-promoting
activity. MCDB 301 contains increased concentrations of calcium chloride and glutamine, and a smaller amount of cysteine than
medium F12. It also has been supplemented with 19 inorganic ions, in addition to selenium and those in medium F12, in order
to insure against possible future deficiencies as chemicals are purified further. A Chinese hamster lung line which will not
grow in MCDB 301 alone will grow when the medium is supplemented either with methylcellulose or with insulin. The growth-promoting
activity is thought to be an impurity shared in common by both substances. The probable “essential” role of impurities in
cellular growth in most synthetic media and the problems involved in attempting to develop a truly “defined” medium are discussed.
This research was supported by Grant No. CA15305 from the National Cancer Institute. 相似文献
8.
Improved medium for clonal growth of human diploid fibroblasts at low concentrations of serum protein 总被引:1,自引:0,他引:1
Wallace L. McKeehan Kerstin A. McKeehan Susan L. Hammond Richard G. Ham 《In vitro cellular & developmental biology. Plant》1977,13(7):399-416
Summary A new medium (MCDB 104) has been developed which will support clonal growth of WI-38 cells at concentrations of serum protein
as low as 25 μg per ml (equivalent to 0.05% serum). The principal factors responsible for reduction of the protein requirement
are: (a) adjustment of all nutrient concentrations in medium F12 to experimentally determined optimum values for WI-38 cells;
(b) supplementation with trace elements; (c) replacement of hypoxanthine and folic acid with adenine and folinic acid; and
(d) coating of the culture surface with polylysine. Individually, many of these modifications exert only a small effect on
cellular growth at reduced protein concentrations, but collectively their effect has been very substantial. Other strains
of fibroblast-like human diploid cells from amniotic fluid, fetal lung and newborn foreskin also will grow at reduced concentrations
of serum protein in the new medium.
This work was supported by Grant No. AG00310 from the National Institute on Aging, and by Contract No. 223-74-1156 from the
Bureau of Biologics, U.S. Food and Drug Administration. 相似文献
9.
Critical adjustment of cysteine and glutamine concentrations for improved clonal growth of WI-38 cells 总被引:2,自引:0,他引:2
Dr. Richard G. Ham Susan L. Hammond Linda L. Miller 《In vitro cellular & developmental biology. Plant》1977,13(1):1-10
Summary Clonal growth of WI-38 cells with a plating efficiency of 45% has been achieved in a synthetic nutrient mixture (MCDB 102)
supplemented with either whole or dialyzed fetal bovine serum. For optimum growth, the concentration of cysteine in the medium
must be adjusted precisely. Deviation by a factor of three in either direction from the optimum concentration (9.0×10−5M) eliminates essentially all clonal growth. A high concentration of glutamine (2.5×10−3M) is also needed for, optimum clonal growth.
Presented in preliminary form at the 26th Annual Meeting of the Tissue Culture Association, June 4, 1975.
This work was supported by Grant No. HD-08181 from the National Institute of Child Health and Human Developement, Grant No.
AG-00310 from the National Institute on Aging, and by Contract No. 223-74-1156 from the Bureau of Biologics, Food and Drug
Administration. 相似文献
10.
Michael H. Simonian Mark L. White David A. Foggia 《In vitro cellular & developmental biology. Plant》1987,23(4):247-252
Summary The life span and growth from clonal density of bovine adrenocortical cell cultures were studied in serum-supplemented medium
and a serum-free defined medium, which supported sustained cell proliferation and steroid production. The total culture life
span was 79 population doublings in serum-supplemented medium with fibroblast growth factor (FGF) and 36 population doublings
in the defined medium without serum. Older passage cell cultures grown in the defined medium progressively lost the ability
to produce 11β- and 21-hydroxylated steroids, which was observed previously for cultures in serum-supplemented medium, and
also had a decline of 17α-hydroxylated steroid production. The cloning efficiency in the defined medium was 12.2% as compared
to 24% in serum-supplemented medium with FGF. Five isolated clonal cell lines grown in the defined medium were characterized
for steroid function in response to steroidogenic agents. All five clonal cell lines had stimulated steroid production with
8-bromo-cAMP, but only four of the clonal lines were stimulated also by adrenocorticotropin. None of the clonal cell lines
produced 11β-, 21- or 17α-hydroxylated steroids in response to treatment with either steroidogenic agent, results that were
similar to data obtained from older mass cultures. The apparent deficiency of the defined medium as compared to serum-supplemented
medium for maximum support of the culture life span and cloning efficiency may be useful in studies of cellular aging and
its relation to differentiated function for this cell culture system.
This study was supported by the Iowa Diabetes and Endocrinology Research Center (grant AM25295 from the National Institutes
of Health, Bethesda, MD). D.A.F. was supported by a National Research Service Award from the National Institutes of Health
(grant HL07485). 相似文献
11.
Paul J. La Rocca James G. Rheinwald 《In vitro cellular & developmental biology. Plant》1985,21(1):67-72
Summary This laboratory recently reported that normal human mesothelial cells require epidermal growth factor (EGF) and hydrocortisone
(HC), in addition to fetal calf serum and a complex defined medium component, in order to grow optimally in surface culture
(9). We report here that this normal cell type also forms large colonies at high efficiency in semi-solid medium, but exhihits
more stringent serum and EGF requirements for anchorage-independent than for surface growth. Mesothelial cells are unable
to divide at all in semi-solid medium with added EGF or with less than 2% serum, whereas they grow slowly but progressively
in surface culture under such conditions. In semi-solid medium containing 20% serum and HC, mesothelial cells are stimulated
to divide by the addition of as little as 30 pg/ml purified EGF. Human urine or male mouse plasma could substitute for purified
EGF, yielding growth commensurate with the levels of EGF in these biological fluids previously measured by others using radioreceptor
and radioimmune assays. Thus growth of mesothelial cells in semi-solid medium can serve as a highly sensitive assay of EGF
biological activity which is unaffected by the presence of serum proteins. In addition, our results demonstrate that fetal
calf serum does not provide mitogenic levels of EGF to cultured cells, raising the question of the identity of plasma and
serum mitogens.
This work was supported by NIH grants RO1 AG02048 and RO1 CA26656 to James G. Rheinwald and by NIH postdoctoral fellowship
F32 AG05303 to Paul J. La Rocca. 相似文献
12.
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
13.
Long-term culture of human endothelial cells 总被引:9,自引:0,他引:9
Portia B. Gordon Ira I. Sussman Victor B. Hatcher 《In vitro cellular & developmental biology. Plant》1983,19(9):661-671
Summary Human umbilical vein endothelial cells can be grown in vitro for 28 passages (CPDL 58) in Medium 199 supplemented with newborn
bovine serum and a partially purified growth factor derived from bovine brain. Newborn bovine serum is superior to fetal bovine
serum for the proliferation of human umbilical vein endothelial cells seeded at low density in the presence of the growth
factor. The endothelial cells, which can be passaged every 7 to 10 d at a 1-to-5 split ratio, retain their morphological and
biochemical characteristics. The proliferation of cells seeded at low density (103/cm2) is proportional to the concentration of the growth factor present in the medium. The growth factor, which has an isoelectric
point between 5.0 and 5.5, can support cell proliferation at reduced serum concentrations; half-maximal growth is achieved
in medium containing the growth factor and 3% serum. The brain endothelial cell growth factor does not stimulate DNA synthesis
significantly in cultures of human skin fibroblasts.
This research was supported by grants from the U.S. Public Health Service (AG 01732, HL 16387, and HL 07080), the Cystic Fibrosis
Foundation, and the New York and American Heart Associations.
Victor B. Hatcher is an Established Fellow of the New York Heart Association and a recipient of the Ann Weinberg Cystic Fibrosis
Research Scholarship Award. 相似文献
14.
Growth of normal human mammary cells in culture 总被引:27,自引:0,他引:27
M. Stampfer R. C. Hallowes A. J. Hackett 《In vitro cellular & developmental biology. Plant》1980,16(5):415-425
Summary Reduction mammoplasty tissue was used to obtain short-term cultures of human epithelial cell populations. Digestion of tissue
with collagenase and hyaluronidase resulted in cell clusters (organoids) resembling ductal and alveolar structures; these
could be separated by filtration from the stromal components. Epithelial outgrowth from these organoids was greatly enhanced
by the addition of conditioned medium from other human epithelial and myoepithelial cell lines. Additionally, the mammary
epithelial growth was stimulated by insulin, hydrocortisone, epidermal growth factor, and steroid hormones. With this enriched
nutritional environment, active cell division could be maintained for 1 to 3 months and cells could be serially subcultured
1 to 4 times.
This research was supported by Grant PDT-72 from the American Cancer Society and Grant CP-70510 from the National Institutes
of Health. 相似文献
15.
Peroxidation products of polyunsaturated fatty acids may cause growth inhibition of cells in culture. This study was carried out to elucidate to what extent peroxidation products may be found in growth media, with and without cells and albumin, using thiobarbituric acid-reactive substances (TBARS) and protein carbonyl groups as measures of peroxidation. The growth of human microvascular endothelial cells was studied as influenced by docosahexaenoic (C22:6, n - 3), arachidonic acid (C20:4. n - 6), and serum albumin. Cell growth was strongly inhibited by the fatty acids, and the inhibition was related to the concentration of TBARS in the medium. Defatted albumin (0.5 g/100 ml) nullified the increase of TBARS in the medium and released the growth inhibition by the fatty acids. With polyunsaturated fatty acids (PUFA) there was a time- and concentration-dependent increase in media TBARS, observed both with and without cells, but the TBARS increase was somewhat greater in the presence of cells. Surprisingly, TBARS in cell-free media also increased somewhat upon increasing the albumin concentration from 0.5 to 5 g/100 ml, and the TBARS increase differed among various preparations of albumin. Unexpectedly, the albumin that had not been defatted gave the lowest TBARS values. The amount of protein carbonyl groups did not differ among various albumin preparations. It is concluded that PUFA may autooxidize in media used for cell cultures, and thereby cause an unspecific growth inhibition, which can be prevented by a low albumin concentration. However, even defatted albumin preparations may contain lipid peroxidation products, the causes and implications of which remain to be elucidated. 相似文献
16.
Kristina Sundqvist Yun Liu Kristina Arvidson Kari Ormstad Lennart Nilsson Rune Toftgård Roland C. Grafström 《In vitro cellular & developmental biology. Animal》1991,27(7):562-568
Summary Human buccal epithelial cells have been reared from explants maintained in supplemented MCDB 153 medium. Primary epithelial
outgrowths show typical structural features and uniformly express keratins; subunit analyses demonstrate expression of keratins
5, 6, 14, 16/17, and 19. The cells exhibit up to 6% colony forming efficiency and divide at about 0.8 population doublings
per day on fibronectin/collagen-coated dishes at clonal density. Studies of markers of proliferation and differentiation in
buccal epithelial cells indicate that epidermal growth factor, cholera toxin, retinoic acid, and pituitary extract each exhibit
a distinctive ability to enhance growth and variably affect cell migration and cell surface area. Transforming growth factorβ-1 inhibits growth and increases surface area without affecting migration, involucrin expression, and cross-linked envelope
formation. Moreover, exposure of cells to fetal bovine serum, the tumor promoting agent 12-O-tetradecanoylphorbol-13-acetate or an elevated Ca2+ concentration (from 0.1 to 1 mM) inhibits growth and induces squamous differentiation as indicated by inhibition of migration, increases in surface area,
involucrin expression, or formation of cross-linked envelopes. The results show that epithelial cells can be reproducibly
derived from explant cultures of human buccal mucosa specimens and the cells transferred under serum-free conditions. Buccal
epithelial cells in culture undergo a pattern of growth and differentiation that mimics parakeratinization in vivo and variably
respond to several agents shown to modulate growth of cells that originate from other types of epithelia.
This work was supported in part by grants from the Swedish National Board of Laboratory Animals, the Swedish Medical Research
Council, the Swedish Natural Science Research Council, the Swedish Fund for Scientific Research Without Animal Experiments,
the Swedish Cancer Society, the Swedish Tobacco Company, and Lions Club International, Djurg?rden, Stockholm, Sweden. 相似文献
17.
18.
Summary Human umbilical vein endothelial cells (HUV-EC) grew rapidly in vitro in medium supplemented with epidermal growth factor,
fetal bovine serum (FBS) and human diploid fibroblast-conditioned medium. The effect of FBS could be replaced partially by
bovine serum albumin, cholesterol, and vitamin E, and completely by further addition of serum dialysate or refeeding every
other day. Among these components, fibroblast-conditioned medium is essential for HUV-EC growth. The HUV-EC were cultured
serially for over 50 population doublings in the 10% FBS containing fibroblast-conditioned medium and for over 40 population
doublings in the serum-free medium. Mitogenic factor(s) present in the medium conditioned by fibroblasts may be related to
endothelial cell growth factor and play an important role angiogenesis and regeneration of vascular endothelium in vitro. 相似文献
19.
Craig D. Albright Raymond T. Jones Eric A. Hudson Joseph A. Fontana Benjamin F. Trump James H. Resau 《Cell biology and toxicology》1990,6(4):379-398
Normal human bronchial epithelial cells (BE) and adenovirus-12 SV40 hybrid virus transformed, non-tumorigenic human bronchial epithelial cells (BEAS-2B) were cultured for 7 days in a serum free hormone supplemented medium. BE cells after 3 days in culture were exposed to conditioned medium (CMt) from confluent BEAS-2B cells. By day 7, CMt-treated BE cells exhibited a lower colony forming efficiency (CFE), fewer cells per colony, and a reduced mitotic index (MI) and BrdU (bromodeoxyuridine) labeling index. CMt also enhanced the expression of a terminally differentiated squamous phenotype in BE cells. Cell free lysates from BEAS-2B cells (CFLt) had effects similar to CMt on the MI and morphology of BE cells. In contrast, CMt and CFLt did not inhibit the growth, or alter the morphology of BEAS-2B cells. Conditioned medium from BE cells (CMn) did not reduce the growth of BEAS-2B cells, and had little effect on the morphology of BE cells. In co-culturesAbbreviations BE
normal bronchial epithelial cells
- BEAS-2B
adenovirus-12 SV40 hybrid virus transformed bronchial epithelial cells
- CMn
conditioned medium from BE cells
- CMt
conditioned medium from BEAS-2B cells
- CFn
cell free lysate from BE cells
- CFLt
cell free lysate from BEAS-2B cells
- BrdU
bromodeoxyuridine
- KGM
keratinocyte growth medium
- TGF-
transforming growth factor type
- NCI-LHC
National Cancer Institute-Laboratory of Human Carcinogenesis
Contribution No. 2801 from the Pathobiology Laboratory, University of Maryland. 相似文献
20.
Hiroyuki Kitajima Hitoshi Niwa 《Biochemical and biophysical research communications》2010,396(4):933-937
The research of human pluripotent stem cells is important for providing the molecular basis for their future application to regenerative medicine. To date, they are usually cultured on feeder cells and passaged by partial dissociation with either enzymatic or mechanical methods, which are problematic for the research using them in the convenience and reproducibility. Here we established a new culture system that allows handling as easily as culturing feeder-free mouse ES cells. This newly developed culture system is based on the combinatorial use of ROCK inhibitor and soluble fibronectin, which enables us to expand human pluripotent stem cells from single cell dissociation on gelatin-coated surface without any feeder cells. In this new culture system, these human pluripotent stem cells can stably grow, even if in clonal density with keeping expression of stem cell markers. These cells also have abilities to differentiate into three germ layers in vivo and in vitro. Furthermore, no chromosomal abnormalities are found even after sequential passage. Therefore this system will dramatically simplify genetic engineering of these human pluripotent stem cells or defining process of their signal pathway. 相似文献