[求助]请教树突状细胞的培养经验 - 经验共享

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标题:[求助]请教树突状细胞的培养经验

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我的培养体系基本上跟上面各位战友的相同，给你发一张俺的DC的靓照，希望对你有帮助

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2012-3-18 12:09

55409619.snap.jpg (56.42 KB)

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我的培养体系基本上跟上面各位战友的相同，给你发一张俺的DC的靓照，希望对你有帮助

—————————————————

你的dc是什么来源的？
那小的亮的是不是淋巴细胞？
你拍照时染色了吗？
望回复！

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你的dc是什么来源的？
那小的亮的是不是淋巴细胞？
你拍照时染色了吗？
望回复！

————————————————————————————————————————————————————————————————

我做的DC共有三种来源，骨髓，外周血和动员后的外周血干细胞。这个是外周血的，是一个移植的供者，拍照时没有染色，我用的是OLYMPUS的倒置显微镜和配套的OLYMPUS的照相系统，可以调节亮度等参数，所以看上去象是染了色的。
至于小的细胞，应该主要是未清除彻底的淋巴细胞为主，也有未诱导成DC的单核细胞。
另外，我的DC还是不成熟的DC，FACS结果显示，CD80的阳性率在80%,CD86在20%左右。
仅供你参考。

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请问有研究DC细胞的表面受体的吗？我想了解DC细胞的表面受体或功能蛋白。先谢谢高手们的指点。

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这里有篇文章，不知对你有用不？
Prognostic Characteristics of Breast Cancer Among
Postmenopausal Hormone Users in a Screened Population
By Karla Kerlikowske, Diana L. Miglioretti, Rachel Ballard-Barbash, Donald L. Weaver, Diana S.M. Buist, William E. Barlow,
Gary Cutter, Berta M. Geller, Bonnie Yankaskas, Stephen H. Taplin, and Patricia A. Carney
Purpose: We determined the risk of breast cancer and
tumor characteristics among current postmenopausal
hormone therapy users compared with nonusers, by duration
of use.
Methods: From January 1996 to December 2000, data
were collected prospectively on 374,465 postmenopausal
women aged 50 to 79 years who underwent screening
mammography. We calculated the relative risk (RR) of
breast cancer (invasive or ductal carcinoma-in-situ) and
type of breast cancer within 12 months of postmenopausal
therapy use among current hormone users with a uterus
(proxy for estrogen and progestin use) and without a uterus
(proxy for estrogen use), compared with nonusers.
Results: Compared with nonusers, women using estrogen
and progestin for > 5 years were at increased risk of
breast tumors of stage 0 or I (RR, 1.51; 95% CI, 1.37 to
1.66), stage II or higher (RR, 1.46; 95% CI, 1.30 to 1.63),
size < 20 mm (RR, 1.59; 95% CI, 1.43 to 1.76), size greater
than 20 mm (RR, 1.24; 95% CI, 1.09 to 1.42), grade 1 or 2
(RR, 1.60; 95% CI, 1.44 to 1.77), grade 3 or 4 (RR, 1.54; 95%
CI, 1.37 to 1.73), and estrogen receptor-positive (RR, 1.72;
95% CI, 1.55 to 1.90). Estrogen-only users were slightly
more likely to have estrogen receptor-positive breast cancer
compared with nonusers (RR, 1.14; 95% CI, 1.06 to 1.23).
Conclusion: Use of estrogen and progestin postmenopausal
hormone therapy for five years or more increased
the likelihood of developing breast cancer, including both
tumors with favorable prognostic features and tumors with
unfavorable prognostic features.
J Clin Oncol 21:4314-4321. © 2003 by American
Society of Clinical Oncology.
POSTMENOPAUSAL HORMONE therapy (HT) has been
associated with increased risk of breast cancer.1-3 Estrogen
plus progestin regimens may be associated with a greater risk of
breast cancer than estrogen-only regimens4-6; however, results
are not consistent or conclusive across studies.2,4-7 It is also
unclear whether HT results in an increased risk of breast cancer
with a favorable prognosis (low stage and grade), less favorable
prognosis (high stage and grade), or both.
Several observational studies have reported that HT users
have smaller8-13 and lower-grade tumors,10,12,14-17 while others
have not shown any influence of HT on tumor size14,15,18-23 or
grade.9,13,18,21,23 Three studies have reported that HT users are
more likely to have estrogen receptor- (ER-) positive tumors.
4,9,24 Several others have reported no association with HT
use and ER status.8,10 –15,18 –20,23 The inconsistent results across
studies may be because many include a small number of women
who had been receiving HT when diagnosed with breast cancer
(n 29 to n 263).4,8,10 –21,23 In addition, many of the studies
did not adequately account for breast cancer surveillance by
screening mammography,4,8 –12,15,16,18 –20,23 which could result in
earlier detection and fewer advanced cancers in HT users. Lastly,
the studies had no information or very limited information on
tumor characteristics associated with type of HT regimen or

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duration of use, which may influence tumor features.4,5,8-24
The Women’s Health Initiative randomized controlled trial
showed that women taking a continuous estrogen and progestin
regimen for more than 4 years had an increased risk of breast
cancer.3 A greater proportion of women on estrogen and progestin
regimens were diagnosed with regional disease (Surveillance,
Epidemiology and End Results [SEER] program staging
system), compared with those using placebo.25 Determining
whether HT has clinically important influences on breast cancer
stage or aggressiveness and cancer detection rates by mammography
in a large, community-based population may contribute to
a woman’s decision to start HT, or to take HT for long periods.
We evaluated the influence of HT on breast cancer risk by
pooling data from six mammography registries where current
HT use, history of hysterectomy, and time between mammography
examinations is prospectively recorded. We inferred that
From the Department of Epidemiology and Biostatistics, and the General
Internal Medicine Section, Department of Veterans Affairs, University of
California, San Francisco, CA; Center for Health Studies, Group Health
Cooperative; Department of Biostatistics, University of Washington, Seattle,
WA; Applied Research Program, DCCPS, National Cancer Institute, Bethesda,
MD; Department of Pathology, and Health Promotion Research,
University of Vermont, College of Medicine, Burlington, VT; Center for
Research Design and Statistical Methods, University of Nevada School of
Medicine, Applied Research Facility, Reno, NV; Department of Radiology,
University of North Carolina, Chapel Hill, NC; Norris Cotton Cancer
Center/Dartmouth-Hitchcock Medical Center/Department of Community
and Family Medicine, Dartmouth Medical School, Lebanon, NH.
Submitted May 21, 2003; accepted September 8, 2003.
This work was supported by a National Cancer Institute–funded Breast
Cancer Surveillance Consortium cooperative agreement (U01CA63740,
U01CA86076, U01CA86082, U01CA63736, U01CA70013, U01CA69976,
U01CA63731, U01CA70040).
Authors’ disclosures of potential conflicts of interest are found at the end
of this article.
Address reprint requests to Karla Kerlikowske, MD, San Francisco
Veterans Affairs Medical Center, General Internal Medicine Section, 111A1,
4150 Clement St, San Francisco, CA 94121; e-mail: kerliko@itsa.ucsf.edu.
© 2003 by American Society of Clinical Oncology.
0732-183X/03/2123-4314/$20.00
4314 Journal of Clinical Oncology, Vol 21, No 23 (December 1), 2003: pp 4314-4321
DOI: 10.1200/JCO.2003.05.151
history of hysterectomy is a proxy for type of HT use based on
clinical practice guidelines for HT that have been in place for
more than 10 years, and recommend estrogen and progestin for
women with a uterus, and estrogen only for women without a
uterus. We report the relative risk (RR) of breast cancer by tumor
characteristics and cancer detection rates by mammography
among HT users with and without a uterus by duration of use,
compared with non-HT users.

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METHODS
Data Sources
Data were pooled from six mammography registries that participate in the
Breast Cancer Surveillance Consortium26 (cuturl('http://breastscreening.cancer.gov'))
funded by the National Cancer Institute: (1) San Francisco Mammography
Registry, San Francisco, CA; (2) Group Health Cooperative, Seattle, WA; (3)
Colorado Mammography Advocacy Project, Denver, CO; (4) Vermont
Breast Cancer Surveillance System, Burlington, VT; (5) New Hampshire
Mammography Network, Lebanon, NH; and (6) Carolina Mammography
Registry, Chapel Hill, NC. These registries collect information on screening
and diagnostic mammography examinations performed in their defined
catchment area. Each mammography registry links women in their registry to
a state tumor registry or regional SEER program that collects populationbased
cancer data. Some registries additionally link to pathology databases.
Each registry obtains annual approval from their institutional review board to
collect registry information. Linkage procedures are performed following
human subjects protocols to maintain participant confidentiality.
Subjects
Our study included postmenopausal women aged 50 to 79 years who
underwent bilateral mammography examination indicated by the radiologist
as being performed for screening, between January 1996 and December
2000. Women entered the study at the time of their screening examination
and were observed for 1 year, or until the next screening examination, to
determine if breast cancer was diagnosed. A woman may have had more than
one mammography examination during the study period, and thus, may have
entered the study more than once (n 192,511). Breast cancer was not
uniformly detected during the follow-up period; rather, detection was highest
immediately following a screening examination. Therefore, all analyses were
performed per mammography examination rather than per person-year of
follow-up. Screening examinations that occurred after December 2000 were
excluded to ensure at least 12 months for reporting cancers to tumor
registries after screening examinations.
Women 55 years and older were assumed to be postmenopausal. Women
aged 50 to 54 years were considered to be postmenopausal if both ovaries
had been removed, if they reported that their periods had stopped permanently,
or if they were taking HT. Premenopausal women aged 50 to 54 years
having regular menstrual periods with no HT use were excluded (66,132;
6%). We also excluded women who self-reported breast augmentation
(1%) or prior diagnosis of breast cancer (3%), and women for whom time
between mammography examinations (4%), family history of breast cancer
(8%), or current HT use (17%) was missing.
Measurements and Definitions
For each woman, demographic information and a self-reported breast
health history were obtained at the time of each screening examination by
completing a survey. The survey includes questions about menopausal status,
family history of breast cancer in a first-degree relative, current HT use,women with a uterus who were using HT were considered to be using
estrogen plus progestin, whereas women without a uterus using HT were
considered to be using estrogen only.
Time between mammography examinations was determined using dates of
prior mammography examinations recorded in each mammography registry,
or self-reported information collected at the screening examination. We used
self-reported data, rather than dates of prior mammography examinations
recorded in a registry, to calculate time between mammography examinations
for 6.7% of nonusers, 3.5% of HT users without a uterus, and 9.3% of
HT users with a uterus.
Women were considered to have breast cancer if reports from a breast
pathology database, SEER program, or state tumor registry showed any
invasive carcinoma or ductal carcinoma in situ (DCIS) through December
2001. Women with lobular carcinoma-in-situ only were not considered to
have cancer. All breast cancers were classified according to the American
Joint Committee on Cancer staging system.27 Invasive cancers were categorized
by tumor size, grade, and ER status. The few women (n 55) with
grade 4 tumors (undifferentiated or anaplastic) were included with the group
of women with grade 3 tumors (poorly differentiated).
Statistical Analysis
We stratified the data into three groups based on self-reported current HT
use and history of hysterectomy: (1) no HT use with or without a uterus, (2)
HT use and no uterus (proxy for estrogen only), and (3) HT use and uterus
(proxy for estrogen and progestin use). Frequency distributions of various
risk factors were determined for women in these three groups. Rates and RRs
were calculated using Poisson regression, adjusting for age as a continuous
variable, family history of breast cancer, examination year, time between
mammography examinations, and mammography registry. We standardized
the rates by taking a weighted average of the rates estimated from the Poisson
regression model for each covariate configuration, weighting by the proportion
of women in the study with that covariate configuration. Thus, the same
weights were used for nonusers, estrogen and progestin users, and estrogenonly
users, resulting in an adjustment to the same population. Adjusted
distributions were standardized to the study population using logistic
regression for ER status, and polytomous regression for other tumor
characteristics. We also adjusted all analyses for race. Since the results were
similar, and to minimize the number of women excluded from analyses due
to missing values, we did not adjust for race in the final analyses.
history of hysterectomy, and time between mammography examinations.
Women were considered to have a family history of breast cancer if they
reported having at least one first-degree relative (mother, sister, or daughter)
with breast cancer. Women were considered to be “current HT users” if they
reported using prescription HT at a screening examination. Three registries
collect information on duration of use, which was categorized as less than 5
and 5 years of use. Two registries collect detailed self-report information
on type of HT regimen for women who self-reported “current HT use.” Of
women who reported current HT use and no history of hysterectomy, 80.4%
reported taking estrogen and progestin, 17.8% reported estrogen only, and
1.8% reported progestin only. Of those who reported current HT use and
history of hysterectomy, 87.3% reported taking estrogen only, 12.0%
reported estrogen and progestin, and 0.7% reported progestin only. Thus,

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Rates of breast cancer (invasive cancer or DCIS diagnosed within 12
months of a screening examination and HT use) and 95% CIs were calculated
per 1000 examinations for the three groups and by decade of age.
Adjusted rates and distributions were calculated for stage (0, I, II, and III
and IV combined), tumor size (invasive cancer 10 mm, 11 to 20 mm,
and 20 mm), tumor grade (1, 2, and 3 and 4 combined), and ER status
(positive and negative).
We calculated RRs comparing those taking estrogen and progestin and
those on estrogen only, with to non-HT users by duration of treatment. We
also calculated RRs comparing the risk of tumor characteristics in estrogen
and progestin users and estrogen-only users, with that of nonusers. We
compared the risk of each more favorable tumor feature (stage 0 or I, size
20 mm, grade 1 or 2, ER-positive) and each less favorable tumor feature
(stage II or higher, size 20 mm, grade 3 or 4, ER-negative). We
dichotomized tumor size at 20 mm and stage at 0 or I since both
parameters are considered early stage disease. To evaluate the impact of
study parameters on our findings, two sensitivity analyses were performed
using the same Poisson regression models described above: 1) for a subset of
women whose time between screens ranged from 9 to 18 months with a
median of 13 months, and 2) including all breast cancers occurring within 24
months of a screening examination.
4315 BREAST CANCER AND POSTMENOPAUSAL HT
We calculated the true-positive and false-negative rates per 1000 examinations
for the three study groups. Adjusted rates were calculated using the
same method as described above. We used simulation to estimate the 95%
confidence intervals, sampling 100,000 values of the regression coefficients
from their joint multivariate normal distribution and calculating the rates for
each sample. We estimated upper and lower limits by the simulated 2.5 and
97.5 percentiles. A screening examination was considered a false-negative
examination if breast cancer was diagnosed within 12 months of a negative
examination (BI-RADS [American College of Radiology, Philadelphia, PA]
assessment of 1, 2, or 3 when associated with short-interval follow-up only
or routine follow-up). A screening examination was considered a truepositive
examination if breast cancer was diagnosed within 12 months of a
positive examination (BI-RADS assessment of 0, 4, 5, or 3 when associated
with a recommendation for immediate follow-up).
RESULTS
A total of 373,265 postmenopausal women underwent
683,435 screening mammography examinations between January
1996 and December 2000, of whom 3,202 developed
breast cancer (2,619 invasive and 583 DCIS) within 12
months of an examination.
HT users were more likely to be younger, white, have had a
previous breast biopsy or surgery, and shorter time period
between mammography examinations and less likely to have a
family history of breast cancer (Table 1).
Among women who developed breast cancer, the mean age at
diagnosis was significantly younger among women using estrogen
and progestin and estrogen only compared with nonusers (61
and 63 years v 66 years; P .001). Overall risk of cancer was
higher among women using estrogen and progestin compared
with nonusers (RR, 1.39; 95% CI, 1.31 to 1.47). There was no
significant increased risk of breast cancer among women using
estrogen alone compared with nonusers (RR, 1.05; 95% CI, 0.99
to 1.12). The rate of cancer increased with age for all three groups
(P .0001) and was higher for each decade of age among women
using estrogen and progestin compared with women using estrogen
only, and compared with nonusers (Table 2).
Rate and Risk of Cancer by Tumor Characteristics and
HT Use

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The rates of stage 0, I, and II tumor; invasive cancer 20
mm; grade 1, 2, and 3 or 4 disease; and ER-positive disease were
higher among women using estrogen and progestin compared
with women using estrogen only, and compared with nonusers
(Table 3).
Rate and risk of breast cancer was higher among women using
estrogen and progestin for 5 years compared with nonusers
(RR, 1.49; 95% CI, 1.36 to 1.63) (Table 4). Rate and risk of
breast cancer was not increased among women using estrogen
and progestin less than 5 years, or among those using estrogen
Table 1. Study Population Characteristics for 374,465 Postmenopausal Women
Who Underwent 683,435 Screening Mammography Examinations Between
1996 and 2000 by HT Use
Hormone Use (% of patients)*
No HT Estrogen Only Estrogen and Progestin
No. of women 213,660 79,216 81,589
No. of examinations 382,435 151,399 149,601
Age, years
50-59 35 49 55
60-69 34 34 32
70-79 31 17 13
Race or ethnicity†
White 83 90 91
Black 9 6 3
Asian 6 3 4
Native American or Alaskan
Native
1 0 0
Other races 1 1 1
Hispanic 3 2 3
Family history of breast cancer 16 14 12
Previous breast biopsy or surgery† 21 27 23
Time between screens, years
1 6270 70
2 2423 23
3-4 7 4 4
4 8 3 3

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Abbreviation: HT, hormone therapy.
*Estrogen-only group: HT users without a uterus. Estrogen and progestin group: HT
users with a uterus. Column percentage calculated from number of examinations.
†Missing data: 13% for race, 14% for ethnicity (Hispanic), and 3% for previous
breast biopsy or surgery.
Table 2. Rate of Cancer and 95% CI Among 374,465 Postmenopausal Women Who Underwent Screening Mammography From 1996 to 2000 by HT Use
Hormone Use*
No HT Estrogen Only Estrogen and Progestin
Rate 95% CI Rate 95% CI Rate 95% CI
No. of examinations 382,435 151,399 149,601
No. of cancers 1,803 624 775
Cancers per 1,000 examinations† 4.3 4.2 to 4.5 4.5 4.3 to 4.8 6.0 5.7 to 6.3
Cancers per 1,000 examinations by age, years‡
50-59 3.3 3.1 to 3.6 3.4 3.2 to 3.8 4.5 4.2 to 4.9
60-69 4.6 4.4 to 5.0 4.8 4.4 to 5.2 6.4 5.9 to 6.9
70-79 5.8 5.5 to 6.6 6.0 5.5 to 6.6 7.9 7.2 to 8.6
Abbreviation: HT, hormone therapy.
*Estrogen-only group: HT users without a uterus. Estrogen and progestin group: HT users with a uterus.
†Adjusted for age, family history of breast cancer, examination year, time between mammography examinations, and mammography registry using Poisson regression,
and standardized to the total population.
‡Adjusted for family history of breast cancer, examination year, time between mammography examinations, and mammography registry using Poisson regression, and
standardized to the total population.
4316 KERLIKOWSKE ET AL
only compared with nonusers, irrespective of duration of use
(Table 4).
The risk of tumors associated with more favorable prognostic
characteristics was higher among women using estrogen and
progestin for 5 years compared with nonusers: 41% for DCIS,
51% for stage 0 or I, 59% for invasive cancer 20 mm or smaller,
60% for grade 1 or 2 disease, and 72% for ER-positive disease
(Table 5). The risk of tumors associated with less favorable
prognostic characteristics was also higher among women using
estrogen and progestin for 5 years compared with nonusers:
51% for invasive cancer; 46% for stage II, III, or IV; 24% for
invasive cancer larger than 20 mm; and 54% for grade 3 or 4
disease (Table 5). Except for the increased risk of an ER-positive
tumor (Table 5), overall cancer risk was not higher among
women using estrogen only as compared with nonusers. In a
sensitivity analysis on a subset of women whose time between
screens was approximately 1 year, results were similar to those
presented in Tables 4 and 5 (data not shown), with risk of breast
cancer higher among women using estrogen and progestin for
5 years compared with nonusers (RR, 1.51; 95% CI, 1.32 to
1.72). A separate sensitivity analysis that allowed 24 months for
cancer to occur after a screening examination, found similar
results to those reported in Tables 4 and 5 (data not shown); risk
of breast cancer was higher among women using estrogen and
progestin for 5 years compared with nonusers (RR, 1.55; 95%
CI, 1.40 to 1.71).

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