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Modified Leukocyte Filter Removes Tumor Cells from the Salvaged Blood.

Mei K, Du L, Yan M, Zhang Z, Zhang F, Gong L, Sun K, Zhang J, Tang Y, Jiang C, Liu J - PLoS ONE (2015)

Bottom Line: Intraoperative blood salvage, an effective blood conservation strategy, has not been applied in onco-surgery, because of potential malignant cell contamination.Invasive growth adherent cells were found in most of unfiltered samples and 67% of the inoculated nude mice developed tumors in LDF-treated sample.Neither adherent cells nor nude mice developed tumors were found in M-LDF-treated samples.

View Article: PubMed Central - PubMed

Affiliation: Department of Medical Oncology, Sichuan Cancer Hospital, Chengdu, Sichuan, China.

ABSTRACT

Background: Intraoperative blood salvage, an effective blood conservation strategy, has not been applied in onco-surgery, because of potential malignant cell contamination. In this study we tested effectiveness of a modified leukocyte depletion filter (M-LDF) for removal of tumor cells.

Materials and methods: The effects of M-LDF and regular LDF on removal of cells (HepG2 cell line) were compared. The safety of M-LDF was tested with blood (collected and washed during onco-surgery), the salvaged blood mixed with tumor cells from the solid tumor of the same patient, or mixed with HepG2 cells (n=30 in each protocol). Cancer cells were identified by flow cytometry, culture and bioassay with and without filtration.

Results: M-LDF removed 5-log of HepG2 and nucleated cells, which was much higher than regular LDF, and cells were destroyed when they passed through M-LDF. Cytokeratin-positive cells in all samples were removed by M-LDF. Invasive growth adherent cells were found in most of unfiltered samples and 67% of the inoculated nude mice developed tumors in LDF-treated sample. Neither adherent cells nor nude mice developed tumors were found in M-LDF-treated samples.

Discussion and conclusion: Since M-LDF can effectively remove and destroy cancer cells in the salvaged blood, it has great potential for clinical application.

No MeSH data available.


Related in: MedlinePlus

Bioassays for tumor development in nude mice.Injection of M-LDF untreated and treated samples into the left axilla of nude mice. With untreated samples, tumors developed (2–3 cm in diameter, A1, Black arrow) and were removed for HE (A2) and immunofluorescent staining (AE1/AE3-PE and DAPI, A3) to confirm human origin. No tumor was found after one year (B1, green arrow) in the M-LDF treated samples. HE staining of tissues from the axilla showed normal cells (B2), and stained negative for AE1/AE3 (B3).
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pone.0130864.g007: Bioassays for tumor development in nude mice.Injection of M-LDF untreated and treated samples into the left axilla of nude mice. With untreated samples, tumors developed (2–3 cm in diameter, A1, Black arrow) and were removed for HE (A2) and immunofluorescent staining (AE1/AE3-PE and DAPI, A3) to confirm human origin. No tumor was found after one year (B1, green arrow) in the M-LDF treated samples. HE staining of tissues from the axilla showed normal cells (B2), and stained negative for AE1/AE3 (B3).

Mentions: M-LDF filtration reduced nucleated cell count from (2.46±1.63) ×109/L to (4.54±12.04) ×104/L (P = 0.000). With unfiltered samples, clones were found in all cultures within 3–5 days, and tumors developed in 20 out of 30 inoculated nude mice (72 days, ranging from 5 to 338 days, Fig 7A1), which was confirmed by pathological and immunofluorescent examinations (Fig 7A2 and 7A3). Neither clones were found in culture and nor tumors developed in mice in the M-LDF-treated group (Fig 7B).


Modified Leukocyte Filter Removes Tumor Cells from the Salvaged Blood.

Mei K, Du L, Yan M, Zhang Z, Zhang F, Gong L, Sun K, Zhang J, Tang Y, Jiang C, Liu J - PLoS ONE (2015)

Bioassays for tumor development in nude mice.Injection of M-LDF untreated and treated samples into the left axilla of nude mice. With untreated samples, tumors developed (2–3 cm in diameter, A1, Black arrow) and were removed for HE (A2) and immunofluorescent staining (AE1/AE3-PE and DAPI, A3) to confirm human origin. No tumor was found after one year (B1, green arrow) in the M-LDF treated samples. HE staining of tissues from the axilla showed normal cells (B2), and stained negative for AE1/AE3 (B3).
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4476620&req=5

pone.0130864.g007: Bioassays for tumor development in nude mice.Injection of M-LDF untreated and treated samples into the left axilla of nude mice. With untreated samples, tumors developed (2–3 cm in diameter, A1, Black arrow) and were removed for HE (A2) and immunofluorescent staining (AE1/AE3-PE and DAPI, A3) to confirm human origin. No tumor was found after one year (B1, green arrow) in the M-LDF treated samples. HE staining of tissues from the axilla showed normal cells (B2), and stained negative for AE1/AE3 (B3).
Mentions: M-LDF filtration reduced nucleated cell count from (2.46±1.63) ×109/L to (4.54±12.04) ×104/L (P = 0.000). With unfiltered samples, clones were found in all cultures within 3–5 days, and tumors developed in 20 out of 30 inoculated nude mice (72 days, ranging from 5 to 338 days, Fig 7A1), which was confirmed by pathological and immunofluorescent examinations (Fig 7A2 and 7A3). Neither clones were found in culture and nor tumors developed in mice in the M-LDF-treated group (Fig 7B).

Bottom Line: Intraoperative blood salvage, an effective blood conservation strategy, has not been applied in onco-surgery, because of potential malignant cell contamination.Invasive growth adherent cells were found in most of unfiltered samples and 67% of the inoculated nude mice developed tumors in LDF-treated sample.Neither adherent cells nor nude mice developed tumors were found in M-LDF-treated samples.

View Article: PubMed Central - PubMed

Affiliation: Department of Medical Oncology, Sichuan Cancer Hospital, Chengdu, Sichuan, China.

ABSTRACT

Background: Intraoperative blood salvage, an effective blood conservation strategy, has not been applied in onco-surgery, because of potential malignant cell contamination. In this study we tested effectiveness of a modified leukocyte depletion filter (M-LDF) for removal of tumor cells.

Materials and methods: The effects of M-LDF and regular LDF on removal of cells (HepG2 cell line) were compared. The safety of M-LDF was tested with blood (collected and washed during onco-surgery), the salvaged blood mixed with tumor cells from the solid tumor of the same patient, or mixed with HepG2 cells (n=30 in each protocol). Cancer cells were identified by flow cytometry, culture and bioassay with and without filtration.

Results: M-LDF removed 5-log of HepG2 and nucleated cells, which was much higher than regular LDF, and cells were destroyed when they passed through M-LDF. Cytokeratin-positive cells in all samples were removed by M-LDF. Invasive growth adherent cells were found in most of unfiltered samples and 67% of the inoculated nude mice developed tumors in LDF-treated sample. Neither adherent cells nor nude mice developed tumors were found in M-LDF-treated samples.

Discussion and conclusion: Since M-LDF can effectively remove and destroy cancer cells in the salvaged blood, it has great potential for clinical application.

No MeSH data available.


Related in: MedlinePlus