Department of Pediatrics, Kyoto University, Kyoto,
Exp Hematol. 2009 Dec;37(12):1400-10. doi: 10.1016/j.exphem.2009.09.006. Epub 2009 Sep 24.
Multipotent germline stem (mGS) cells derived from neonatal mouse testis, similar to embryonic stem (ES) cells, differentiate into various types of somatic cells in vitro and produce teratomas after inoculation into mice. In the present work, we examined mGS cells for hematopoietic progenitor potential in vitro and in vivo.
mGS cells were differentiated on OP9 stromal cells and induced into Flk1(+) cells. Flk1(+) cells were sorted and replated on OP9 stromal cells with various cytokines and emerging hematopoietic cells were analyzed for lineage marker expression by fluorescein-activated cell sorting, progenitor activity by colony assay, and stem cell transplantation assay.
mGS cells, like ES cells, produce hematopoietic progenitors, including both primitive and definitive erythromyeloid, megakaryocyte, and B- and T-cell lineages via Flk1(+) progenitors. When transplanted into the bone marrow (BM) of nonobese diabetic/severe combined immunodeficient (NOD/SCID) gammac(null) mice directly, mGS-derived green fluorescent protein (GFP)-positive cells were detected 4 months later in the BM and spleen. GFP(+) donor cells were also identified in the Hoechst33342 side population, a feature of hematopoietic stem cells. However, these mGS-derived hematopoietic cells did not proliferate in vivo, even after exposure to hematopoietic stressors, such as 5-fluorouracil (5FU) injection or serial transplantation.
mGS cells produced multipotent hematopoietic progenitor cells with myeloid and lymphoid lineage potential in vitro and localized in the BM after intra-BM injection but, like ES cells, failed to expand or show stem cell repopulating ability in vivo.
从新生小鼠睾丸中分离出的多能生殖系干细胞（mGS）类似于胚胎干细胞（ES），在体外可分化为各种类型的体细胞，并在接种入小鼠后产生畸胎瘤。本研究旨在检测 mGS 细胞在体外和体内的造血祖细胞潜能。
mGS 细胞在 OP9 基质细胞上分化，并诱导成 Flk1(+)细胞。Flk1(+)细胞被分选出来，在含有各种细胞因子的 OP9 基质细胞上重新铺板，通过荧光激活细胞分选分析出现的造血细胞的谱系标记表达，通过集落形成实验分析祖细胞活性，通过干细胞移植实验分析。
mGS 细胞与 ES 细胞一样，通过 Flk1(+)祖细胞产生造血祖细胞，包括原始和终末红系、巨核细胞以及 B 和 T 细胞谱系。当直接将 mGS 衍生的绿色荧光蛋白（GFP）阳性细胞移植到非肥胖型糖尿病/严重联合免疫缺陷（NOD/SCID）γc(null)小鼠的骨髓（BM）中时，4 个月后在 BM 和脾脏中检测到 GFP 阳性细胞。GFP(+)供体细胞也在 Hoechst33342 侧群中被鉴定出来，这是造血干细胞的一个特征。然而，这些 mGS 衍生的造血细胞在体内并没有增殖，即使在暴露于造血应激物（如 5-氟尿嘧啶（5FU）注射或连续移植）后也没有增殖。
mGS 细胞在体外产生具有髓系和淋巴系潜在分化能力的多能造血祖细胞，并在 BM 内注射后定位于 BM 中，但与 ES 细胞一样，在体内无法扩增或显示出干细胞再殖能力。
