Migration assay on primary culture isolated from patient's primary breast cancer tissue

ED Yuliana, A Bustami A Bustami, HP Tantry

Abstract

Background: Migration is an essential component of breast cancer metastasis, which study
has been concentrated on culture of established breast cancer cell lines that do not accurately
represent the sophistication and heterogeneity of patient's breast cancer. An attempt to
perform migration assay using Boyden Chamber Assay (BCA) on primary culture originating
from patient's breast cancer tissue was developed to accommodate upcoming study of breast
cancer migration in lndonesian patients.
Methods: Pathologically proven primary breast cancer tissue samples were obtained from
Ciptomangunkusumo Hospital during core (n=4) and incisional (n=3) biopsies of stage llA
up to stage lllA breast cancer patients. Following biopsy, the breast cancer tissue samples
underwent processings to isolate the cancer cells. These cancer cells were -then resuspended
within Dulbecco's modified Eagle's medium (DMEM) ahd cultured in 12-well plate. The growth
of primary culture were observed and compared between the core biopsy and the incisional
biopsy specimens. Optimization of BCA method was later performed to investigate the
migration of the breast cancer primary culture towards different experirnental conditions, which
were control, Fetal Bovine Serum (FBS), and Stromal Derived Factor-l (SDF-1). Two different
number of breast cancer cells were tested for the optimization of the BCA, which were 1 x 105
and3x105cells.
Results: None of the culture performed on core biopsy specimens grew, while one out of
three incisional biopsy specimens grew until confluence. The one primary culture that grew
was later assesed using BCA to assess its migration index towards different experimental
conditions. Using 1 x 10s breast cancer cells in the BCA , the result of the absorbance level of
migrated cells showed that the migration towards SDF-1 (0.529) nearly doubled the migration
towards controlmedium (0.239) and FBS (0.209). Meanwhile, the absorbance levelwas simiiar
between the control medium (1.050), FBS (1 .103), and SDF-1 (1 .104) when the same test was
run with 3 x 105 breast cancer cells.
Discussion: Breast cancer tissue collected using incisional biopsies hao better chance to
grow in primary culture than those collected using core biopsies, possibly due to the difference
in the amount of tissue extracted. Consistent with previous research, patient's breast cancer
cells was shown to migrate more towards the chemokine ligand SDF-1, as compared to control
medium and FBS. The number of breast cancer cells run in the BCA also affected the result of
the migration assay, where 1 x 10s cells showed a superiority as compared to 3 x 105 cells in
showing the migration difference between the different experimental conditions.
MKA, Volume 37, Nomor.Supl. 2, November 2014
ED Yulian dkk
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Migration assay on primary culture isolated from patient's primary breast cancer tissue
Conclusions: This study encourages and facilitates future research to study migration the intrinsic characteristic of individualized patient's breast cancer as well as to assess the change in migration characteristic of breast cancer cells after an in vivo treatment in human.

Keywords

breast cancer; primary culture; migration; Boyden chamberAssay; sDF_1 B3

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