Establishment and characterization of transplantable tumor line (KB) and cell lines (KB-P and KB-D8) from a rat thymus-derived dendritic cell sarcoma

Abstract

A transplantable tumor line (KB) was established in syngeneic rats from a naturally occurring sarcoma that had arisen in the thymus of a 24-month-old male F344 rat. Further, a cell line (KB-P) was induced from KB and a cloned cell line (KB-D8) was isolated from KB-P. The primary thymic tumor and KB tumors showed heterogeneous histological growth patterns such as sheet-like, ill-defined bundle, fascicular and interwoven fashions, consisting of spindle cells, oval cells and histiocytic large round cells. Immunohistochemically, neoplastic cells in KB tumors and KB-P and KB-D8 cultures reacted to vimentin and were labeled with antibodies of OX6 (for rat major histocompatibility complex class-II antigens), ED5 (for rat follicular dendritic cells; FDCs) and RED-1 (for interdigitating dendritic cells) in varying degrees, indicating that neoplastic cells exhibited the immunophenotypes of rat dendritic cells. In addition, neoplastic cells were immunoreactive to ED1 (for rat monocytes/macrophages) and ED2 (for rat tissue macrophages), and also showed positive reactions to histiocytic lysosomal enzymes such as acid phosphatase and non-specific esterase. Ultrastructurally, neoplastic cells had cell surface projections, cisterna-like structures and variously developed lysosomes in the cytoplasm. Based on these findings, the present tumor was regarded as dendritic cell-derived sarcoma capable of expressing macrophage-like and histiocytic nature. A reverse-transcription polymerase chain reaction method revealed that the addition of lipopolysaccharide dose dependently increased the expression of mRNA of transforming growth factor-beta1, a proinflammatory factor, in KB-D8 cells. The transplantable line (KB) and cell lines (KB-P and KB-D8) may become useful tools for studying the histogenesis and pathobiological functions of dendritic cells.

Characterization of a Rat Subcutaneous Malignant Fibrous Histiocytoma and Its Tumor Lines, with Reference to Histiocytic Features

“…Serial transplantations then were made at 4-to 5-week intervals until the fourth generation by implanting a tumor tissue fragment. 37,38,40 In each passage, 2 or 3 F344 rats were used. The homotransplantable tumor line was named KJ.…”

“…38,40 Tissue sections were stained by hematoxylin and eosin (HE), periodic acid-Schiff (PAS), Watanabe's silver impregnation, azanMallory, alcian blue (pH 2.5) and phosphotungustic acid hematoxylin (PTAH) methods. Frozen sections from the formalin-fixed tissues were stained with oil red O.…”

“…33,38,40 RT-PCR was conducted as described previously. 33,38,40 From 5 mg of total RNA, the single-strand cDNA was synthesized with Super Script Preamplification System (GIBCO BRL), and cDNAs were amplified by PCR with Taq DNA polymerase (Takara Shuzou, Kyoto, Japan) and each of the specific primers (sense and antisense) for TNF-a or b-actin (control). The following conditions were used for the amplification for TNF-a: 32 cycles of 2 minutes of denaturation at 94uC, 1 minute of annealing at 55uC, and 0.5 minutes of synthesis at 42uC.…”

“…Toxicologic pathologists who perform the evaluations in these studies require detailed knowledge concerning the characteristics of spontaneously occurring neoplastic lesions. Because F344 rats are an inbred strain with a genetically homogeneous nature, we have utilized this strain to establish tumor lines (homotransplantable and cultured cell lines) from spontaneous tumors, 33,[37][38][39][40] in order to clarify the biologic behavior, morphogenesis, and paraneoplastic syndromes associated with these tumors. In the present study, we succeeded in establishing a homotransplantable tumor line (KJ) in syngeneic rats from a spontaneous MFH found in the subcutaneous tissue of an aged male F344 rat.…”

Cisplatin-Induced Renal Interstitial Fibrosis in Neonatal Rats, Developing as Solitary Nephron Unit Lesions

“…The methods have been described elsewhere (Yamate et al, 2002a;Ide et al, 2003). Briefly, after pretreatments applied for each primary antibody (Table 1) to inactivate endogenous peroxidase, the deparaffinized sections were treated with 5% skimmed milk for 30 minutes, and incubated with each primary antibody for 14-24 hours.…”