Wiktor-Jedrzejczak W, Ahmed A, Szczylik C
Exp Hematol. 1981 Sep;9(8):835-48.
The number of adherent colonies grown and the type of their growth were influenced by both the serum and the medium used for the culture. The test results were obtained using either Dulbecco MEM, TC-199 or NCTC-109 together with 10% fetal calf serum and 10% horse serum (FCS/HS). Murine marrow cultures made in these conditions gave the linear relationship between the number of cells explanted (in the range 5 X 10(3) cells/ml to 5 X 10(6) cells/ml) nd the number of macroscopic adherent colonies grown. The number of colonies reached plateau by day 9 of culture and did not increase for up to 90 days of culture. Adherent colony forming cells disappeared from the liquid phases of such cultures within 3 days. Adherent layer formed had the capacity to support maintenance of spleen colony forming units and produced colony stimulating factor (CSF). This CSF supported 4-day granulocyte and macrophage colony formation in liquid cultures but not 7-day granulocyte-macrophage colony formation in semisolid medium. Therefore, the described culture system allowed to quantitatively grow cells responsible for some properties attributed to hemopoietic microenvironment (HM), possessed defined growth characteristics and may be utilized for further in vitro studies of HM.
所生长的贴壁集落数量及其生长类型受到培养所用血清和培养基的影响。使用杜尔贝科改良伊格尔培养基（Dulbecco MEM）、TC - 199或NCTC - 109以及10%胎牛血清和10%马血清（FCS/HS）获得了测试结果。在这些条件下进行的小鼠骨髓培养，在接种细胞数量（范围为5×10³个细胞/毫升至5×10⁶个细胞/毫升）与所生长的宏观贴壁集落数量之间呈现线性关系。集落数量在培养第9天达到平台期，并且在长达90天的培养期内不再增加。贴壁集落形成细胞在这类培养物的液相中3天内消失。形成的贴壁层有能力支持脾集落形成单位的维持并产生集落刺激因子（CSF）。这种CSF在液体培养中支持4天的粒细胞和巨噬细胞集落形成，但在半固体培养基中不支持7天的粒细胞 - 巨噬细胞集落形成。因此，所描述的培养系统能够定量培养负责造血微环境（HM）某些特性的细胞，具有明确的生长特征，可用于进一步的HM体外研究。
