Differentiation of canine bone marrow cells with hemopoietic characteristics from an adherent stromal cell precursor.
Differentiation of canine bone marrow cells with hemopoietic characteristics from an adherent stromal cell precursor.
复制标题
具有造血特性的犬骨髓细胞从贴壁基质细胞前体的分化。
DOI：
10.1073/pnas.92.3.748
复制
发表时间：
1995
影响因子：
11.1
通讯作者：
Deeg,HJ
中科院分区：
文献类型：
--
作者：
Huss,R;Hong,DS;McSweeney,PA;Hoy,CA;Deeg,HJ
从犬长期骨髓培养物中建立基质细胞系，通过有限稀释法克隆，并维持在基质细胞条件培养基中。这些细胞贴壁生长，在标准条件下（在20-30%条件培养基中;汇合率，70-90%）保持稳定的生长速率和形态，并在长期骨髓培养中支持造血。在外源性重组犬干细胞因子（rcSCF）的存在下，圆形细胞从粘附层，脱离，并作为活的漂浮细胞留在培养物中。当培养物在没有rcSCF的情况下生长至> 90%汇合时，也出现圆形漂浮细胞。圆形细胞比贴壁细胞小，表达CD 34，显示嗜碱性血浆，c-kit、MHC-II类标志物和髓样标志物染色阳性。在标准的集落形成试验中，分离的细胞产生粒细胞-巨噬细胞集落形成单位（CFU-GM）、成纤维细胞集落形成单位（CFU-F）和不太明确的集落形成单位。此外，在长期培养的同种异体饲养细胞上，这些细胞产生造血集落。引人注目的是，分化是可逆的：当非贴壁细胞以较低密度重悬于含血清培养基中时，它们重新贴壁并生长至汇合，此时圆形细胞再次分离。当置于克隆测定中时，来自该第二周期的分离细胞主要产生CFU-F和很少的CFU-GM。这些结果表明，一些成纤维样基质细胞具有分化为具有造血特性的细胞的潜力。这些观察结果为成年犬骨髓基质中存在造血祖细胞的静止前体提供了证据。
Stromal cell lines were established from canine long-term marrow cultures, cloned by limiting dilution, and maintained in stromal cell-conditioned medium. These cells grew adherent, maintained stable growth rate and morphology under standard conditions (in 20-30% conditioned medium; confluency, 70-90%), and supported hemopoiesis in long-term marrow cultures. In the presence of exogenous recombinant canine stem cell factor (rcSCF), round cells developed from the adherent layer, detached, and remained in culture as viable floating cells. Round floating cells also appeared when cultures were grown to > 90% confluency without rcSCF. Round cells were smaller than adherent cells, expressed CD34, showed basophilic plasma, and stained positive for c-kit, MHC-class II markers, and myeloid markers. In standard assays for colony formation, the detached cells produced granulocyte-macrophage colony-forming units (CFU-GM), fibroblast colony-forming units (CFU-F), and less well-defined colony-forming units. In addition, on allogeneic feeder cells in long-term cultures, these cells generated hemopoietic colonies. Strikingly, the differentiation was reversible: when nonadherent cells were resuspended at lower density in serum-containing medium, they reattached and grew to confluence when, once again, round cells detached. Detached cells from this secondary cycle produced mainly CFU-F and few CFU-GM when placed in clonal assays. These results suggest that some fibroblast-like stromal cells have the potential to differentiate into cells with hemopoietic characteristics. These observations provide evidence for the existence of a quiescent precursor of hemopoietic progenitors in the bone marrow stroma of the adult dog.