Most studies of cancer stem cells (CSC) involve the inoculation of cells from human tumors into immunosuppressed mice, preventing an assessment on the immunologic interactions and effects of CSCs. In this study, the investigators examined the vaccination effects produced by CSC-enriched populations from histologically distinctmurine tumors after their inoculation into different syngeneic immunocompetent hosts. Enriched CSCs were immunogenic and more effective as an antigen source than unselected tumor cells in inducing protective antitumor immunity.Immune sera from CSC-vaccinated hosts contained high levels of IgG which bound to CSCs, resulting in CSC lysis in the presence of complement.CTLs generated from peripheral blood mononuclear cells or splenocytes harvested from CSC-vaccinated hosts were capable of killing CSCs in vitro. Mechanistic investigations established that CSC-primed antibodies and T cells were capable of selective targeting CSCs and conferring antitumor immunity.

To assess the feasibility of generating CSC-loaded DC vaccines for clinical use, the investigators will harvest peripheral blood and tumor specimen from patients with Breast Cancer. The investigators will purify T, B cells and generate DCs from the PBMCs of the Breast Cancer patient.On the other hand, investigators will isolate ALDHhigh and ALDHlow tumor cells from the tumor specimen of the Breast Cancer patient using a similar protocol as investigators reported .

Aim 1: To demonstrate, in vitro, the relative cellular anti-Breast Cancer CSC immunity induced by Breast Cancer CSC-DC primed cytotoxic T cells.

Aim 2: To determine, in vitro, specific binding and lysis of Breast Cancer CSCs by antibodies produced by purified B cells from PBMCs stimulated with Breast Cancer CSC-DC.

Inclusion Criteria:

• The patient is ≥ 30 years of age at the time the informed consent to screening has been obtained;
• The patient has one of the following histologically confirmed breast cancer subtypes:

Estrogen receptor and/or progesterone positive tumor; Human epidermal growth factor receptor 2 (HER2)-overexpressing breast cancer; HER2-negative breast cancer.

-- The patient shows normal organ function according to the following parameters(as measured within six weeks prior to treatment allocation):

• Hemoglobin: Within normal range according to institutional standards;
• Absolute leukocyte count: Within normal range according to institutional standards;
• Absolute lymphocyte count: Within normal range according to institutional standards;
• Platelet count: Within normal range according to institutional standards;
• Alanine aminotransferase: ≤ 2.5 x Upper Limit of Normal (ULN);
• Aspartate aminotransferase: ≤ 2.5 x ULN;
• Total bilirubin: ≤ 1.5 x ULN. In the case of known Gilbert's syndrome ≤ 2 x ULN;
• Serum creatinine: 1.5 x ULN;
• Calculated creatinine clearance: > 50 mL/min .

Exclusion Criteria:

• The patient has inflammatory breast cancer, which is defined as clinically significant erythema of the breast and/or documented dermal lymphatic invasion.
• Diagnosis established by incisional biopsy.
• Prior and concomitant neoadjuvant anti-breast-cancer treatments such as chemotherapy, immunotherapy / biological response modifiers, endocrine therapy, and radiotherapy, unless authorized specifically by the protocol.
• level 3 hypertension;
• severe coronary disease;
• myelosuppression;
• respiratory disease;
• brain metastasis;
• chronic infections

The patients who unresectable after Cryotherapy