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1.
Primary and long term epithelial cell cultures from human fetal normal colonic mucosa 总被引:3,自引:0,他引:3
Primary and passaged cultures of normal colon epithelial cells, derived from human fetuses (13 to 17 wk of conceptual age) have been established. These cultures have been passaged 16 times thus far. The cultures have been initiated and maintained in medium consisting of 50% Dulbecco's minimum essential medium and 50% Ham's F12 medium and supplemented with antibiotics (penicillin, 100 U/ml; streptomycin, 100 micrograms/ml); ascorbic acid, 40 micrograms/ml; L-isoleucine, 50 micrograms/ml; epidermal growth factor, 20 ng/ml; insulin, 5 micrograms/ml; cholera toxin, 5 ng/ml; transferrin, 1 microgram/ml; fetal bovine serum (10%); and HEPES, 25 mM final concentration, and incubated at 37 degrees C in humidified gas containing 5% CO2: 95% air. The cellular and subcellular characteristics of primary and passaged cultures were defined using light microscopy and scanning and transmission electron microscopy. The cells exhibited microvilli on cell surfaces and showed junctional complexes and interdigitations between cells. Indented nuclei with dense chromatin and marginated heterochromatin, numerous mitochondria, rough endoplasmic reticulum, polysomes, and extensive Golgi zones were conspicuous. Also, periodic acid Schiff's reagent-positive staining of the cells suggests the active synthesis of complex mucopolysaccharides in the cytoplasm. 相似文献
2.
R. E. Gibson-D’Ambrosio M. Samuel C. C. Chang J. E. Trosko S. M. D’Ambrosio 《In vitro cellular & developmental biology. Plant》1987,23(4):279-287
Summary Epithelial cell cultures were prepared from normal human fetal kidney and established in long-term culture. The growth characteristics
and production of keratin, and alkaline phosphatase (AP) and gamma-glutamyl transpeptidase (GGT) activities were compared
in a modified minimal essential medium (mMEM),d-valine-containing modified alpha-MEM (mALPHA) andl-valine mALPHA. The mean number of cumulative population doublings (CPDL) was significantly (P<0.001) enhanced with thel-valine mALPHA (40.8 CPDL) over that achievable in mMEM (14.2 CPDL) ord-valine mALPHA (18.3 CPDL) media. In all three media, greater than 95% of the cells in culture produced keratin throughout
the life span of these cultures. Surface-associated fibronectin was absent in these cell cultures. AP and GGT activities increased
as a function of subpassage and time in culture, with the greatest activity in thel-valine mALPHA. The expression of these renal cell-associated functions suggests that these cells in culture are proximal
tubule epithelial cells. The conditions and procedures described in this paper can provide a human kidney epithelial cell
culture system for studying human renal function, metabolism, cytotoxicity, genotoxicity, and transformation.
Research was supported by a NIEHS (ES 3101) grant to S. M. D’Ambrosio and a NCI grant (CA21104) to J. E. Trosko. 相似文献
3.
E. M. Earley S. C. Finley W. H. Finley L. Y. F. Hsu H. J. Kim J. C. Petricciani P. C. Vinson 《In vitro cellular & developmental biology. Plant》1976,12(9):639-642
Summary Cell cultures were established from the biopsies of lung, skin and kidney from each of nine human fetuses, and chromosome
analyses were performed on material through the fifth subculture. Kidney cell cultures generally showed a higher level of
polyploidy than lung or skin. The frequencies of hyperdiploid cells and those with structural abnormalities were consistent
with the low levels found in cultures of human lymphocytes. The data provide a normal cytogenetic baseline for human fetal
material which may be useful in a variety of studies.
Supported by Food and Drug Administration contracts FDA 74-51 and 74-52.
Authors are listed alphabetically. 相似文献
4.
Epithelial cell cultures from the colon of the suckling rat 总被引:2,自引:0,他引:2
Summary Epithelial cells from the colon of suckling rats have been propagated in vitro. The colons were excised and cut longitudinally.
The epithelial sheets were peeled off and dissociated in 0.1% trypsin solution at 25°C for 10 min. The first cell suspension
was discarded and the remaining fragments trypsinized again for an additional 20 min. The dissociated cells were washed and
cultured. Forty-eight hours later, several epithelial colonies consisting of closely packed polygonal cells were formed. Transmission
and scanning electron microscope examination of the colonies showed numerous regularly spaced microvilli on the surface and
tight junctions and desmosomes between adjacent cells. Immunocytochemical studies with antiserum prepared against the brush-border
membrane of the colonic epithelium showed specific staining of the epithelial colonies.
Epithelial colonies were subcultured by the penicylinder method. Although the subcultured cells retained their epithelial
characteristics, the proliferative activity of the cells gradually decreased. Currently, efforts are being made to determine
the optimum nutritional requirements of the primary and low-passage cultures. 相似文献
5.
A method for the rapid establishment of normal adult mammalian colonic epithelial cell cultures 总被引:6,自引:0,他引:6
Alda Vidrich Rajeswari Ravindranath Kianbanoo Farsi Stephan Targan 《In vitro cellular & developmental biology. Plant》1988,24(3):188-194
Summary Normal colonic epithelial cell cultures of mammalian origin are required to facilitate the study of both normal cellular functions
as well as pathogenesis of certain (human) colonic diseases. To date, little information is available regarding the growth
requirements of colonic epithelial cells in culture of eitehr animal or human origin. Such data would enable the development
of a long-term culture system for these cells. In this study, we present methodology that results in the establishment of
homogeneous cultures of adult rabbit colonic epithelial reproducibly, quickly, and in quantity. The epithelial nature of the
cultures is unambiguously established by intermediate filament typing using antikeratin antibodies. Such culutres can now
be used for a variety of functional studies as well as to investigate the growth requirements of colonic epithelial in culture.
This work was supported by the Blinder Foundation for Crohn’s Disease Research, Harbor UCLA IBD Center (AM 36200) and grant
AM 27806 from the National Institutes of Health, Bethesda, MD. 相似文献
6.
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
7.
Summary Serial passage cultures of colonic epithelial cells from young rats have been maintained for more than 6 months in Eagle's
minimum essential medium buffered with HEPES (25 mM) and supplemented with 2.5% fetal bovine serum, 0.5 μg/ml insulin, 5.0 μg/ml transferrin, and antibiotics. The cells proliferated
in this medium with a population doubling time of approximately 53 h. The cells retained differentiated morphology as evidenced
by secretory activity and the presence of secretory granules, microvilli, tonofilaments, and desmosomal junctions. Further
cells at the fourth passage had normal karyotypes with 42 chromosomes and exhibited anchorage dependent growth. High concentrations
of fetal bovine serum (10 to 15%) exerted toxic effects on the colonic epithelial cell cultures.
Supported by National Cancer Institute Contract N01-CP-75914. 相似文献
8.
Dharam P. Chopra Richard L. Shoemaker Gregory W. Taylor Patricia A. Mathieu 《In vitro cellular & developmental biology. Animal》1991,27(1):13-20
Summary Cultures of normal human tracheal gland epithelial cells that exhibit functional differentiation have been propagated in serum-free
medium supplemented with insulin (5 μg/ml), epidermal growth factor (10 ng/ml), hydrocortisone (0.5 μg/ml), and bovine pituitary
extract (25 μg/ml). The cells retain many characteristics of epithelial cells including microvilli on cell surfaces, desmosomes
between cells, and tonofilaments in the cytoplasm. In addition, they exhibit keratin-positive titers and react positively
with Peanut agglutinin, which is specific for the disaccharide β-d-galactose-(l→ 3)N-acetyld-galactosamine, a major component of mucin glycoprotein. The cells also exhibit normal Cl− channel activity which was enhanced by the cAMP agonist Forskolin. The major component of the cellular secretion was hyaluronic
acid; approximately 10% of the void volume material was resistant to hyaluronidase and may contain material similar to mucin
glycoprotein. Some of the cell cultures have been maintained in serum-free conditions for 6 to 7 passages. This model will
be important to study regulation of ion-channel activities and mucous glycoprotein secretion and to compare such regulations
with the tracheal mucosal epithelial cells already established.
This research was supported by USPHS grants HL 41979 and HL 33142 from the National Heart, Lung and Blood Institutes. 相似文献
9.
Michael H. Simonian Michael W. Capp Mark C. Templeman Alizabeth C. Chang 《In vitro cellular & developmental biology. Plant》1987,23(1):57-62
Summary The human fetal adrenal cortex is one of the largest fetal organs and synthesizes precursors for placental estrogen production
as part of the feto-placental unit. The factors controlling the rapid growth of the human fetal adrenal cortex during the
second and third trimesters are not known. Placental regulation of the growth of human fetal adrenocortical cell cultures
from second trimester fetuses was studied. A placental-derived mitogenic factor (PDMF) was detected in tissue homogenates
of 14 to 22 week human placentas and stimulated adrenocortical cell number and [3H]thymidine incorporation into DNA 5–8 fold. PDMF has been partially purified by ammonium sulfate precipitation and anion
exchange chromatography. PDMF is a heat sensitive protein with disulfide bonds required for activity. The growth stimulation
by PDMF was significantly greater than that for basic or acidic fibroblast growth factor by 25–50% and epidermal growth factor
by 3–4 fold. The placental hormones, progesterone, estriol, estradiol, placental lactogen and chorionic gonadotropin, either
alone or in combination did not stimulate fetal adrenocortical cell growth, except for a 41% cell number increase by progesterone.
Platelet-derived growth factor and insulin-like growth factors I and II were not mitogenic for these cells. These results
show that the placenta contains a potent growth factor for human fetal adrenocortical cell cultures. This implies a direct
role for the placenta in control of this fetal organ’s growth, which would make the human feto-placental unit a bi-directional
relationship.
This research was supported by Grants HD15882 and HD21798 from the National Institutes of Health, Bethesda, MD. This work
was presented in part at the 68th Annual Meeting of The Endocrine Society, Anaheim, CA, June 1986.
Editor’s Statement This report describes a potentially new relationship between placenta and the regulation of growth of fetal
adrenalcortical cells. Since these cells produce several of the essential hormones influencing fetal development, characterization
of this factor(s) could provide important insights into the process of differentiation. David A. Sirbasku 相似文献
10.
Eileen A. Friedman Paul J. Higgins Martin Lipkin Hiromi Shinya Alvin M. Gelb 《In vitro cellular & developmental biology. Plant》1981,17(7):632-644
Summary Human colonic epithelial cells from three classes of benign tumors have been reproducibly cultured free of fibroblasts for
8 wk using a supplemented Medium 199 (M 199S). The cultured colonic cells were identified as epithelial by the presence of
junctional complexes (tight junctions, gap junctions, and desmosomes), a brush border on the apical surface, keratin fibrils,
and by both a close-packed columnar or cuboidal morphology and the capability to transport water and ions to form hemicysts.
Colony formation was initiated by groups of epithelial cells, not by single cells, and was inhibited by cocultivation with
either lethally irradiated 3T3 cells or human diploid fibroblasts. Enhancement of epithelial colony formation was observed
following culture on nonadherent, “floating” substrates compared with substrates attached directly to the bottom of the culture
dish.
Replication of epithelial cells in M 199S from the class of benign colonic tumors least prone to malignancy, the tubular,
was significantly enhanced by epidermal growth factor (EGF). In contrast, EGF did not stimulate the growth of cells in M 199S
from the other classes of benign tumors, the villotubular and the villous, which exhibit more malignant potential. These data
imply that premalignant colonic epithelial cells lose responsiveness to growth modulation by EGF as they progress toward frank
carcinoma.
This study was supported by NCI Contract N01-CP43366 to M. L. and NCI Grant 1-R26-CA 28822 to E. F. 相似文献
11.
Lynne P. Rutzky Beppino C. Giovanella Baldwin H. Tom Celia I. Kaye Philip D. Noguchi Barry D. Kahan 《In vitro cellular & developmental biology. Plant》1983,19(2):99-107
Summary An epithelial cell line, LS123, was established in 1974 from the second in a series of three primary colonic tumors resected
from a Caucasian female. The cell line is aneuploid, releases low concentrations of carcinoembryonic antigen (CEA), fails
to grow progressively in nude mice, and forms colonies only in enriched semisolid medium developed for tumor stem cells. LS123
cells grow on confluent cell monolayers and in either low serum or serum-free medium. In the chick embryonic skin assay, LS123
cells grew as a well-differentiated abnormal colonic epithelium with little mitotic activity but with some indication of invasion.
On floating collagen gels LS123 cells formed a one to three-cell-layer-thick undifferentiated epithelial sheet. The apparent
low invasiveness of the cells of this line is supported by the patient's history of three primary colon tumors without systemic
metastases during the past 30 yr. Therefore, although LS123 cells possess several properties associated with neoplasia, they
have little invasive potential. Thus, LS123 cells may represent an important model for the study of human colon cancer.
Presented in part at the 33rd Annual Meeting of the Tissue Culture Association, San Diego, CA, June 6–10, 1982.
The work has been partially supported by U.S. Public Health Service Grants CA23871 (L. P. R.), CA24024 and RCDAK04-CA00579
(B. H. T.), and CA27124 and CA22370 (B. D. K.); the latter was awarded through the National Large Bowel Cancer Project. 相似文献
12.
B. Delhotal F. Lemonnier M. Couturier C. Wolfrom M. Gautier A. Lemonnier 《In vitro cellular & developmental biology. Plant》1984,20(9):699-706
Summary The effect of fructose as a substitute for glucose in cell culture media was investigated in human skin fibroblast and liver
cell cultures. Cells were grown for between 2 and 10 days in identical flasks in four different media, containing 5.5, mmol·1−1 and 27.5 mmol·I−1 glucose and fructose, respectively. In the presence of fructose, cell growth was stimulated, but less in liver cells than
fibroblasts. At Day 6, increases were observed in [3H]thymidine incorporation, protein levels, and amino acid consumption, and a reduction was noted in ATP levels. In media containing
5.5, mmol·1−1 glucose or fructose, consumption of fructose was four times lower than that of glucose at Day 3 and did not rise until Day
6. In fructose media, the lactate production was very low (four to five times less than that of glucose) and the pH values
were always higher. Some findings were different for the fibroblasts and liver cells, owing to the specific characteristics
of these two cell types in culture; this applied especially to the effects of glucose and fructose concentrations of 27.5
mmol·1−1. Several possible explanation for the stimulation of cell growth in fructose medium were discussed.
This work was supported by grants for the Institut National de la Santé et de la Recherche Médicale (ATP 82-79-114) and the
Unité d'Enseignement et de Recherche, Le Kremlin-Bicêtre, Université Paris-Sud (C. R. 848). 相似文献
13.
M. Jorissen J. Vereecke E. Carmeliet H. Van den Berghe J.-J. Cassiman 《The Journal of membrane biology》1990,117(2):123-130
Summary The patch-clamp technique was used to characterize ion channels in the apical membranes of cultured human nasal epithelial cells, dissociated from fetal nasal mucosa and from adult nasal polyps. Outward-rectifying chloride channels were found in 4.3% of the cell-attached patches from fetal cells (n=258) and in 3.1% of the patches from adult cells (n=320). After exeision the number of patches containing active chloride channels increased threefold to 13% of the patches from the fetal cells and 10% from adult cells. The single-channel conductance at 0 mV in symmetrical 150mm NaCl solutions was 24.3 ±0.9 pS (n=28) and 26.0 ± 1.2 pS (n=30), respectively, in adult and fetal cells and showed outward rectification in the potential range from –80 to +80 mV. In fetal cells as well as in adult cells the channels were anion selective, and were almost impermeable for larger anions and monovalent cations. In cell-free patches the channels were Ca2+ independent. In most of the channels the open probability was voltage independent and high (±0.86); in 20% of the channels, however, the open probability increased with depolarization. In conclusion, fetal nasal epithelial cells contain chloride channels in their apical membranes with singlechannel properties and regulatory mechanisms similar to those found in cells from adults. 相似文献
14.
Albert S. Herring Ratna Raychaudhuri Susan P. Kelley P. Thomasiype 《In vitro cellular & developmental biology. Plant》1983,19(7):576-588
Summary A number of liver epithelial cell cultures were established from 10 to 12-d-old sucklings, 6 to 8-wk-old young adults, or
from 2 to 18-month-old partially hepatectomized rats. Improvements in the methods for cell isolation and culture yielded replicating
cells from every experiment and they were maintained for different periods with regular passages. The proliferative potential
in vitro of adult rat liver cells could be increased if the rats were subjected to partial hepatectomy before cell isolation.
In the early passages, the majority of the cells were found to have a true diploid karyotype as studied by the Giemsa-banding
technique. Under the culture conditions described, a very high percentage of cells remained in the diploid range for, in most
cases, at least 4 months and in some cases for up to 6 months. Afterward, the karyotype was unstable, but no “crisis” period
was seen before the cells became aneuploid. Until this time, the growth characteristics of the cells also followed a normal
pattern showing density dependent inhibition of division and a lack of markers associated with malignancy. The cultured liver
cells exhibited a number of liver specific properties when they were maintained as a confluent monolayer. The early passages
of diploid epithelial cell cultures derived from normal and regenerating rat liver are good models for studies of the regulation
of cell division and the changes that are related to carcinogenesis.
Research Sponsored by the National Cancer Institute, DHHS, under Contract N01-CO-23909 with Litton Bionetics, Inc. The contents
of this publication do not necessarily reflect the views or policies of the Department of Health and Human Services, nor does
mention of trade names, commercial products, or organizations imply endorsment by the U.S. Government. 相似文献
15.
Ning Yi Yap Teng Aik Ong Christudas Morais Jayalakshmi Pailoor Glenda C. Gobe Retnagowri Rajandram 《Cell biology international》2019,43(6):715-725
Renal cell carcinoma (RCC) is one of the most lethal urogenital cancers and effective treatment of metastatic RCC remains an elusive target. Cell lines enable the in vitro investigation of molecular and genetic changes leading to renal carcinogenesis and are important for evaluating cellular drug response or toxicity. This study details a fast and easy protocol of establishing epithelial and fibroblast cell cultures or cell lines concurrently from renal cancer nephrectomy tissue. The protocol involves mechanical disaggregation, collagenase digestion and cell sieving for establishing epithelial cells while fibroblast cells were grown from explants. This protocol has been modified from previous published reports with additional antibiotics and washing steps added to eliminate microbial contamination from the surgical source. Cell characterisation was carried out using immunofluorescence and quantitative polymerase chain reaction. Eleven stable epithelial renal tumour cell lines of various subtypes, including rare subtypes, were established with a spontaneous immortalisation rate of 21.6% using this protocol. Eight fibroblast cell cultures grew successfully but did not achieve spontaneous immortalisation. Cells of epithelial origin expressed higher expressions of epithelial markers such as pan‐cytokeratin, cytokeratin 8 and E‐cadherin whereas fibroblast cells expressed high α‐smooth muscle actin. Further mutational analysis is needed to evaluate the genetic or molecular characteristics of the cell lines. 相似文献
16.
Summary The growth of short-term primary cultures of endometrial epithelium has been studied using Feulgen microspectrophotometry.
A gradual increase in the number of polyploid nuclei up to 64C has been observed and is associated with a decline in the growth
capacity of the cultures. The specific mechanism(s) of this polyploidization is not known. 相似文献
17.
Han Deng Sumona Mondal Shantanu Sur Craig D. Woodworth 《Journal of cellular physiology》2019,234(6):7683-7694
Cervical cancer is a major public health problem and research using cell culture models has improved understanding of this disease. The human cervix contains three anatomic regions; ectocervix with stratified squamous epithelium, endocervix with secretory epithelium, and transformation zone (TZ) with metaplastic cells. Most cervical cancers originate within the TZ. However, little is known about the biology of TZ cells or why they are highly susceptible to carcinogenesis. The goal of this study was to develop and optimize methods to compare growth and differentiation of cells cultured from ectocervix, TZ or endocervix. We examined the effects of different serum-free media on cell attachment, cell growth and differentiation, and cell population doublings in monolayer culture. We also optimized conditions for organotypic culture of cervical epithelial cells using collagen rafts with human cervical stromal cells. Finally, we present a step-by-step protocol for culturing cells from each region of human cervix. 相似文献
18.
Robert E. Priest Jean H. Priest Jessie F. Moinuddin Demetrios S. Sgoutas 《In vitro cellular & developmental biology. Plant》1979,15(2):142-147
Summary Two of the distinguishable cell classes subcultured from human amniotic fluid were examined for their capability to produce
human chorionic gonadotropin (hCG) as determined by radioimmunoassay. The class that predominates in most cultures used for
prenatal genetic diagnosis, previously termed AF (for amniotic fluid), secretes hCG into the culture medium. Dermal fibroblasts
do not, nor does another type of cultured cell from amniotic fluid, previously termed F because of a resemblance to fibroblasts.
Primary AF cultures produce more hCG than do subcultures. Evidence that this hormone is intact hCG is provided by its immunoreactivity
with antisera raised against the β-subunit and against the intact molecule of hCG. Furthermore, a dose-response curve for
hormone in culture medium is parallel to that of highly purified intact hCG. It is postulated that AF cultures are derived
from fetal membranes and retain properties of trophoblast.
Research supported by Grand HD 11379 from the National Institutes of Health. 相似文献
19.
Enrichment and characterization of clonogenic epithelial cells from adult rat liver and initiation of epithelial cell strains 总被引:8,自引:0,他引:8
Kazunori Furukawa Tomiko Shimada Patricia England Yohichi Mochizuki Gary M. Williams 《In vitro cellular & developmental biology. Plant》1987,23(5):339-348
Summary A highly efficient method is described for obtaining prolifertive epithelial cells from adult rat livers for the reproducible
establishment of liver epithelial cell strains. When cells were isolated from livers of 10-to 15-wk-old male Fischer 344 rats
by a collagenase-perfusion method, collected by centrifugation at 50×g for 5 min, and cultured in Williams' medium E containing fetal bovine serum and dexamethasone, colonies of epithelial cells
different in size and morphology from hepatocytes were obtained. Sequential perfusion with collagenase and dispase yielded
numerous epithelial cell colonies. When isolated cells were fractionated by differential centrifugation, the great majority
of hepatocytes were sedimented at 50 ×g for 1 min, whereas many non-hepatocytic cells remiined in the supernatant and could be sedimented by a second centrifugation
at 50×g for 5 min. Culture of the two fractions revealed that almost all the epithelial cell colonies were derived from cells in
the non-hepatocytic cell fraction. The epithelial cells were cytochemically negative for γ-glutamyl transpeptidase activity,
whereas an increase in the activity was detected in hepatocytes with duration in culture. Ultrastructural characteristics
of hepatocytes were not found in the cells of newly established cell strains. These results suggest that adult rat liver epithelial
cells propagable in culture were derived from a cell type other than the hepatocyte. 相似文献
20.
The human oviduct is known as a functional site for gamete transportation, retention, fertilization and zygote development. Previous studies have shown that human oviductal epithelial cell cultural medium (OECCM) has a positive effect on prolongation of sperm motility for some cryopreserved human sperm without cryodamage. However, for most cryopreserved sperm, OECCM could not improve their survival prolongation. In this study, we assessed the influence of human OECCM on the motility longevity of cryopreserved human sperm with an in vitro incubation method. 相似文献