Carter R F, Kruth S A, Valli V E, Dubé I D
Department of Pathology, University of Toronto, Ontario, Canada.
Exp Hematol. 1990 Oct;18(9):995-1001.
We established and maintained long-term cultures of marrow from normal dogs and dogs with lymphoma or leukemia by single inoculations of mononuclear cell suspensions. Media containing only horse sera (as opposed to horse and fetal calf sera) and catalase (for antioxidative effect) supported improved culture viability, as indicated by increased recovery of progenitor cells (granulocyte-macrophage colony-forming units, CFU-GM) and the release of abundant erythroid cells in the cultures for up to 3 weeks. CFU-GM were maintained for at least 3-4 weeks of culture. Culture appearance, cell counts, and assays of CFU-GM were used to compare the culture kinetics of tumor-involved marrow to normal marrow specimens. Cultures of marrow with extensive tumor involvement tended to be less viable, apparently due to a relative lack of competent progenitors. To investigate whether canine long-term marrow culture provided a purging effect similar to the loss of tumor cells noted in human long-term cultures of marrow from patients with chronic myelogenous leukemia (CML) or acute myelogenous leukemia (AML), we established long-term marrow cultures from 28 dogs with histologically confirmed untreated lymphoma or leukemia. Eleven of these dogs had cytogenetically marked tumor cells in the marrow at the initiation of culture. In six dogs with lymphoma and one dog with acute monocytic leukemia (AMoL) French-American-British classification (FAB) M4 leukemia, we could detect no cytogenetic evidence for persistence of the tumor clones in individually plucked or pooled CFU-GM grown from 3-week-old long-term cultures. In one case of AML (FAB M2), 80% of CFU-GM recovered from long-term cultures at 4 weeks still contained an extra metacentric marker chromosome associated with the continued presence of the leukemic clone in the cultures. Our documentation of a purging effect for some tumors supports the use of this canine model system in the investigation of autologous marrow transplantation with long-term cultured cells for humans with lymphoma and leukemia.
我们通过单核细胞悬液的单次接种，建立并维持了正常犬以及患有淋巴瘤或白血病犬的长期骨髓培养物。仅含马血清（与马血清和胎牛血清相对）和过氧化氢酶（用于抗氧化作用）的培养基支持了更好的培养活力，这表现为祖细胞（粒细胞 - 巨噬细胞集落形成单位，CFU - GM）回收率增加，并且在长达3周的培养中培养物中有大量红系细胞释放。CFU - GM在培养中维持至少3 - 4周。培养物外观、细胞计数以及CFU - GM测定用于比较肿瘤累及骨髓与正常骨髓标本的培养动力学。肿瘤广泛累及的骨髓培养物活力往往较低，显然是由于相对缺乏有功能的祖细胞。为了研究犬长期骨髓培养是否能提供类似于在慢性粒细胞白血病（CML）或急性粒细胞白血病（AML）患者的人长期骨髓培养中所观察到的肿瘤细胞丢失的清除效应，我们从28只经组织学证实未经治疗的淋巴瘤或白血病犬建立了长期骨髓培养物。其中11只犬在培养开始时骨髓中有细胞遗传学标记的肿瘤细胞。在6只患有淋巴瘤的犬和1只患有急性单核细胞白血病（AMoL）（法国 - 美国 - 英国分类法（FAB）M4白血病）的犬中，我们在从3周龄长期培养物中单独挑取或汇集的CFU - GM中未检测到肿瘤克隆持续存在的细胞遗传学证据。在1例AML（FAB M2）中，从4周龄长期培养物中回收的CFU - GM的80%仍含有与培养物中白血病克隆持续存在相关的额外中着丝粒标记染色体。我们对某些肿瘤清除效应的记录支持了在研究将长期培养细胞用于淋巴瘤和白血病患者的自体骨髓移植中使用这种犬模型系统。
