Lazarus H M, Haynesworth S E, Gerson S L, Caplan A I
Department of Medicine, Ireland Cancer Center of University Hospitals of Cleveland, Case Western Reserve University, Ohio 44106, USA.
J Hematother. 1997 Oct;6(5):447-55. doi: 10.1089/scd.1.1997.6.447.
The purpose of this study was to compare the ability to collect human bone marrow-derived mesenchymal (stromal) progenitor cells (MPC) from bone marrow versus peripheral blood hematopoietic progenitor cell (PBPC) collections using in vitro and in vivo assays. Ten milliliter samples of PBPC collections mobilized from 11 patients undergoing autotransplants using chemotherapy followed by G-CSF 5-10 micrograms/kg were evaluated using in vitro and in vivo assays for hematopoietic progenitors and MPCs. Additionally, 10 ml samples of unstimulated bone marrow aspirates as well as PBPC collected after mobilization using G-CSF 10 micrograms/kg obtained from 3 normal, histocompatible allogeneic donors were analyzed for hematopoietic progenitors and MPCs. The MPCs were isolated and culture-expanded as adherent cells in vitro and subsequently tested for the capacity to differentiate into mesenchymal phenotypes in vivo using calcium hydroxyapatite porous ceramic cubes implanted s.c. in athymic mice. Demineralized sections of these cubes were analyzed histologically for the appearance of bone and cartilage. Seven autotransplant subjects with cancer received G-CSF after chemotherapy administration, whereas 4 cancer patients and all 3 normal donors received G-CSF alone as the mobilizing regimen. For the autologous PBPC collections and the normal marrow aspirations, median hematopoietic progenitor content was in the normal range for our institution. MPCs were detected in in vitro cultures and as bone-positive ceramic cubes in samples of all 3 allogeneic donor bone marrows but in none of the 14 autologous and 6 allogeneic PBPC collections. In conclusion, MPCs could not be recovered in PBPC collections obtained from either normal donors or patients who underwent PBPC collections after mobilization therapy but could be obtained routinely from bone marrow samples. Although the role of transplanted MPCs is an area of clinical investigation, this study points out a fundamental differences in the population of cells transplanted after collection from bone marrow versus peripheral blood.
本研究的目的是通过体外和体内试验,比较从骨髓与外周血造血祖细胞(PBPC)采集中获取人骨髓间充质(基质)祖细胞(MPC)的能力。对11例接受化疗后使用5 - 10微克/千克G - CSF进行自体移植的患者所采集的10毫升PBPC样本,采用体外和体内试验评估造血祖细胞和MPC。此外,对3名正常、组织相容性匹配的同种异体供体在使用10微克/千克G - CSF动员后采集的未刺激骨髓抽吸物以及PBPC的10毫升样本进行造血祖细胞和MPC分析。MPC作为贴壁细胞在体外分离并培养扩增,随后使用皮下植入无胸腺小鼠的羟基磷灰石多孔陶瓷立方体,测试其在体内分化为间充质表型的能力。对这些立方体的脱矿切片进行组织学分析,观察骨和软骨的出现情况。7例癌症自体移植受试者在化疗后接受G - CSF,而4例癌症患者和所有3名正常供体仅接受G - CSF作为动员方案。对于自体PBPC采集和正常骨髓抽吸物,造血祖细胞含量中位数在本机构的正常范围内。在所有3名同种异体供体骨髓样本的体外培养物中检测到MPC,并在陶瓷立方体样本中检测到骨阳性,但在14例自体和6例同种异体PBPC采集中均未检测到。总之,在正常供体或动员治疗后进行PBPC采集的患者所获得的PBPC采集中无法回收MPC,但可常规从骨髓样本中获得。尽管移植MPC的作用是一个临床研究领域,但本研究指出了从骨髓与外周血采集中获取的移植细胞群体的根本差异。