Nexcelom Bioscience
Cancer Cell Lines Cellometer Applications

Analyzing NCI-60 Cancer Cell Lines

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Introduction to NCI-60 Human Cancer Cell Lines

The late 1980s saw the development and creation of the National Cancer Institute (NCI) 60 human tumor cell line anticancer drug discovery project. This project was intended to screen thousands of pharmacological compounds for anticancer activity, thus replacing earlier cumbersome en vivo models. The 60 created cell lines represent nine human cancers: breast, central nervous system, colon, kidney, leukemia, lung, melanoma, ovary, and prostate. Since its inception, the testing of anticancer drugs using the NCI-60 panel has generated the most extensive cancer pharmacology database in the world.3 It has also provided invaluable knowledge in genome mutations, drug – tumor interactions and general tumor biology.

Here we have performed Cellometer image cytometry on all NCI-60 cancer cell lines. Every cell sample was individually stained with trypan blue, imaged and analyzed using the Cellometer T4. Each sample was assessed for viability and cell size. The ability to accurately measure cell viability and cell size are two important components in determining the functionality anticancer compounds. Cell death, or reduced cell size in certain tumor cell lines could be an indicator of compound efficacy.

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Measuring Cell Concentration and Viability

Simple, User-friendly Cellometer Procedure

After staining the cells with trypan blue, and loading the sample into the counting chamber slide, the slide is inserted into the Cellometer instrument which automatically captures and analyzes multiple cell images. Within seconds, the result table displays the live, dead, and total cell concentrations, as well as the percent viability.

Using Cellometer Mini, Auto 1000, Auto T4, Auto 2000, K2 and Vision

Simple, User-friendly Procedure

1. Pipette 20µl

2. Insert Slide

3. Select Assay & Click Count

4. Results in 30 seconds!

Trypan Blue Protocol

  1. Dilute the stock (0.4 %) with PBS to 0.2 %.
  2. Filter the trypan blue with 0.2 micron filter
  3. Mix the cell suspension at 1:1 with 0.2 % trypan blue
  4. Load the counting chamber slide into the Cellometer and analyze

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Measuring Cell Size of NCI-60 Cancer Cell Lines

The ability to measure cell size has been a valuable tool for researchers examining the effects of pharmacological agents on cancer cells. Many of the NCI-60 cancer cell lines have distinctly different morphological characteristics and cell sizes.

The Cellometer uses an "Accurate Cell Membrane Outline Algorithm" that produces the cell membrane outline around each cell. When the counting is complete, a green outline can be seen around each counted cell. This sophisticated algorithm allows for consistent identification of the desired cell population and provides reliable and accurate cell counts, concentration, viability and cell size measurements. After each count a Cell Diameter Histogram is automatically generated representing the cell size distribution of the counted population.

786-O cells

786-O cell size histogram

Trypan Blue stained 786-O cells

Trypan Blue stained 786-O cells

Cellometer counted 786-O cells

Cellometer counted 786-O cells

In this experiment average cell diameter for 786-O cells was measured to be 17.6 microns. From the cell diameter histogram, we can estimate that approximately 60% of the cells had a cell diameter that was between 16.6 and 18.6 microns.

SW-620 cells

SW-620 cell size histogram

Trypan Blue stained SW-620 cells

Trypan Blue stained SW-620 cells

Cellometer counted SW-620 cells

Cellometer counted SW-620 cells

In comparison to 786-O cells, the average diameter of SW-620 cells was measured to be 11.1 microns. Looking at the cell diameter histogram, approximately 75 % of the cells had a cell diameter that was between 10.5 and 12.0 microns.

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NCI-60 Human Cancer Cell Lines

Using the Cellometer Auto T4, we imaged and counted every NCI-60 cancer cell line, obtaining the cell concentration, viability, and cell size. Shown here, are the enlarged representative images of each cell line. Since all of the cells were stained with trypan blue, we show the stained trypan blue image as well as the Cellometer counted image.

The cells are organized alphabetically by the organ or origin.

Breast

Shown, a stained trypan blue image and a Cellometer counted image. Counted live cells are outlined in green while the dead trypan blue positive cells are outlined in red.

BT-549 cells

NCI-60 BT-549 human breast cells

Hs 578T

NCI-60 Hs 578T human breast cells

MCF7

NCI-60 MCF7 cells

MDA-MB-231

NCI-60 MDA-MB-231 cells

MDA-MB-468

NCI-60 MDA-MB-468 cells

T-47D

NCI-60 T-47D cells

Central Nervous System

Shown, a stained trypan blue image and a Cellometer counted image. Counted live cells are outlined in green while the dead trypan blue positive cells are outlined in red.

SF-268

NCI-60 SF-268 cells

SF-295

NCI-60 SF-295 cells

SF-539

NCI-60 SF-539 cells

SNB-19

NCI-60 SNB-19 cells

SNB-75

NCI-60 SNB-75 cells

U251

NCI-60 U251 cells

Colon

Shown, a stained trypan blue image and a Cellometer counted image. Counted live cells are outlined in green while the dead trypan blue positive cells are outlined in red.

Colo205

NCI-60 Colo205 cells

HCC-2998

NCI-60 HCC-2998 cells

HCT-116

NCI-60 HCT-116 cells

HCT-15

NCI-60 HCT-15 cells

HT29

NCI-60 HT29 cells

KM12

NCI-60 KM12 cells

SW-620

NCI-60 SW-620 cells
 

Kidney

Shown, a stained trypan blue image and a Cellometer counted image. Counted live cells are outlined in green while the dead trypan blue positive cells are outlined in red.

786-O

NCI-60 786-O cells

A498

NCI-60 A498 cells

ACHN

NCI-60 ACHN cells

CAKI-1

NCI-60 CAKI-1 cells

RXF 393

NCI-60 RXF 393 cells

SN12C

NCI-60 SN12C cells

TK-10

NCI-60 TK-10 cells

UO-31

NCI-60 UO-31 cels

Leukemia

Shown, a stained trypan blue image and a Cellometer counted image. Counted live cells are outlined in green while the dead trypan blue positive cells are outlined in red.

CCRF-CEM

NCI-60 CCRF-CEM cells

HL-60

NCI-60 HL-60 cells

K-562

NCI-60 K-562 cells

MOLT-4

NCI-60 MOLT-4 cells

RPMI-8226

NCI-60 RPMI-8226 cells

SR

NCI-60 SR cells

Lung

Shown, a stained trypan blue image and a Cellometer counted image. Counted live cells are outlined in green while the dead trypan blue positive cells are outlined in red.

A549

NCI-60 A549 cellls

EKVX

NCI-60 EKVX cells

HOP-62

NCI-60 HOP-62 cells

HOP-92

NCI-60 HOP-92 cells

NCI-H226

NCI-60 NCI-H226 cells

NCI-H23

NCI-60 NCI-H23 cells

NCI-H322M

NCI-60 NCI-H322M cells

NCI-H460

NCI-60 NCI-H460 cells

NCI-H522

NCI-H522

 

Melanoma

Shown, a stained trypan blue image and a Cellometer counted image. Counted live cells are outlined in green while the dead trypan blue positive cells are outlined in red.

LOX-IMVI

NCI-60 LOX-IMVI cells

M14

NCI-60 M14 cells

MALME-3M

NCI-60 MALME-3M cells

MDA-MB-435

NCI-60 MDA-MB-435 cells

SK-MEL-2

NCI-60 SK-MEL-2 cells

SK-MEL-28

NCI-60 SK-MEL-28 cells

SK-MEL-5

NCI-60 SK-MEL-5 cells

UACC-257

NCI-60 UACC-257 cells

UACC-62

NCI-60 UACC-62 cells

 

Ovarian

Shown, a stained trypan blue image and a Cellometer counted image. Counted live cells are outlined in green while the dead trypan blue positive cells are outlined in red.

IGROV1

NCI-60 IGROV1 cells

NCI/ADR-RES

NCI-60 NCI/ADR-RES cells

OVCAR-3

NCI-60 OVCAR-3 cells

OVCAR-4

NCI-60 OVCAR-4 cells

OVCAR-5

NCI-60 OVCAR-4 cells

OVCAR-8

NCI-60 OVCAR-4 cells

SK-OV-3

NCI-60 SK-OV-3 cells

 

Prostate

Shown, a stained trypan blue image and a Cellometer counted image. Counted live cells are outlined in green while the dead trypan blue positive cells are outlined in red.

DU-145

NCI-60 DU-145 cells

PC-3

NCI-60 PC-3 cells

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List and Description of NCI-60 Cancer Cell Lines

Name Species Organ of Origin Doubling Time (Hrs) Disease Culture
BT549 Human Breast 53.9 Ductal Carcinoma Adherent
HS 578T Human Breast 53.8 Carcinoma Adherent
MCF7 Human Breast 25.4 Adenocarcinoma Adherent
MDA-MB-231 Human Breast 41.9 Adenocarcinoma Adherent
MDA-MB-468 Human Breast 62 Adenocarcinoma Adherent
T-47D Human Breast 45.5 Ductal Carcinoma Adherent
SF268 Human CNS 33.1 Anaplastic Astrocytoma Adherent
SF295 Human CNS 29.5 Glioblastoma-Multiforme Adherent
SF539 Human CNS 35.4 Glioblastoma Adherent
SNB-19 Human CNS 34.6 Glioblastoma Adherent
SNB-75 Human CNS 62.8 Astrocytoma Adherent
U251 Human CNS 23.8 Glioblastoma Adherent
Colo205 Human Colon 23.8 Dukes' type D, Colorectal adenocarcinoma Adherent & Suspension
HCC 2998 Human Colon 31.5 Carcinoma Adherent
HCT-116 Human Colon 17.4 Carcinoma Adherent
HCT-15 Human Colon 20.6 Dukes' type C, Colorectal adenocarcinoma Adherent
HT29 Human Colon 19.5 Colorectal adenocarcinoma Adherent
KM12 Human Colon 23.7 Adenocarcinoma, Grade III Adherent
SW620 Human Colon 20.4 Adenocarcinoma Adherent
786-O Human Kidney 22.4 renal cell adenocarcinoma Adherent
A498 Human Kidney 66.8 Adenocarcinoma Adherent
ACHN Human Kidney 27.5 renal cell adenocarcinoma Adherent
CAKI Human Kidney 39 clear cell carcinoma Adherent
RXF 393 Human Kidney 62.9 Poorly Differentiated Hypernephroma Adherent
SN12C Human Kidney 29.5 Carcinoma Adherent
TK-10 Human Kidney 51.3 Spindle Cell carcinoma Adherent
UO-31 Human Kidney 41.7 Carcinoma Adherent
CCRF-CEM Human Leukemia 26.7 acute lymphoblastic leukemia Suspension
HL-60 Human Leukemia 28.6 acute promyelocytic leukemia Suspension
K562 Human Leukemia 19.6 chronic myelogenous leukemia Suspension
MOLT-4 Human Leukemia 27.9 acute lymphoblastic leukemia Suspension
RPMI-8226 Human Leukemia 33.5 plasmacytoma, myeloma Suspension
SR Human Leukemia 28.7 Large Cell, Immunoblastic Suspension
A549 Human Lung 22.9 Adenocarcinoma Adherent
EKVX Human Lung 43.6 Adenocarcinoma Adherent
HOP-62 Human Lung 39 Adenocarcinoma Adherent
HOP-92 Human Lung 79.5 Large Cell, Undifferentiated Adherent & Suspension
NCI-H226 Human Lung 61 squamous cell carcinoma; mesothelioma Adherent
NCI-H23 Human Lung 33.4 adenocarcinoma; non-small cell lung cancer Adherent
NCI-H322M Human Lung 35.3 Small Cell Bronchioalveolar Carcinoma Suspension
NCI-H460 Human Lung 17.8 carcinoma; large cell lung cancer Adherent
NCI-H522 Human Lung 38.2 adenocarcinoma; non-small cell lung cancer Adherent
LOX IMVI Human Melanoma 20.5 Malignant Amelanotic melanoma Semi-Adherent
M14 Human Melanoma 26.3 malignant melanoma Adherent
MALME-3M Human Melanoma 46.2 malignant melanoma Adherent & Suspension
MDA-MB-435 Human Melanoma 25.8 Adenocarcinoma Adherent
SK-MEL-2 Human Melanoma 45.5 malignant melanoma Adherent
SK-MEL-28 Human Melanoma 35.1 malignant melanoma Adherent
SK-MEL-5 Human Melanoma 25.2 malignant melanoma Adherent
UACC-257 Human Melanoma 38.5 malignant melanoma Adherent
UACC-62 Human Melanoma 31.3 malignant melanoma Adherent
IGROV1 Human Ovary 31 Cystoadenocarcinoma Adherent
OVCAR-3 Human Ovary 34.7 Adenocarcinoma Adherent
OVCAR-4 Human Ovary 41.4 Adenocarcinoma Adherent
OVCAR-5 Human Ovary 48.8 Adenocarcinoma Adherent
OVCAR-8 Human Ovary 26.1 Adenocarcinoma Adherent
SK-OV-3 Human Ovary 48.7 Adenocarcinoma Adherent
NCI-ADR-RES Human Ovary 34 Adenocarcinoma Adherent
DU145 Human Prostate 32.3 Carcinoma Adherent
PC-3 Human Prostate 27.1 grade IV, adenocarcinoma Adherent

Compiled data from multiple sources 1, 2, 3

Download the full NCI-60 cancer cell line data list, which includes the cell size measurements of all 60 cell lines.

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Conclusions

The Cellometer line of instruments is capable of measuring the cell concentration, viability, and cell size of all 60 NCI cancer cell lines. The innovative "Accurate Cell Membrane Outline Algorithm" assures that all cell samples are de-clustered and counted properly. The ability of the Cellometer instruments to accurately and consistently measure cell concentration, viability, and cell size provides researchers with a reliable tool in their cancer research.

Give us a call Give us a call 978-327-5340. Experienced Nexcelom Applications Specialists are available 8:30am to 5:30pm EST to assist with selection of a Cellometer system.
Ask an Applications Specialist

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Publications Using Cellometer Counting for NCI-60 Cancer Cell Lines

  • Tedeschi PM, Markert EK, Gounder M, et al. (2013) Contribution of serine, folate and glycine metabolism to the ATP, NADPH and purine requirements of cancer cells. Cell Death & Disease 4:e877.
  • Dolfi SC, Chan LL, Qiu J, et al. (2013) The metabolic demands of cancer cells are coupled to their size and protein synthesis rates. Cancer & Metabolism 1(1):20. 

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Additional Citations

  1. Jerry M. Collins, Developmental Therapeutics Program NCI/NIH. http://dtp.nci.nih.gov/
  2. DTP, DCTD Tumor Repository. A Catalog of in Vitro Cell Lines, Transplantable Animal and Human Tumors and Yeast. The Division of Cancer Treatment and Diagnosis (DCTD), National Cancer Institute. 2013
  3. Abaan OD, Polley EC, Davis SR, et al. (2013) The exomes of the NCI-60 panel: a genomic resource for cancer biology and systems pharmacology. Cancer Research.

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