Although embryonic stem (ES) cells can differentiate into germ cells, little is known about the influence of culture media on this process. We investigated the effect of two culture media on the capacity of ES cells to differentiate into germ cells using embryoid body (EB) and monolayer culture protocols. Germ cell differentiation was induced in mouse ES cells under four experimental conditions: EB/Dulbecco's modified Eagle's medium (EB/DMEM), EB/knockout Dulbecco's modified Eagle's medium (EB/KO-DMEM), monolayer/Dulbecco's modified Eagle's medium (monolayer/DMEM), and monolayer/knockout Dulbecco's modified Eagle's medium (monolayer/KO-DMEM). After incubation for 6 days, quantitative real-time polymerase chain reaction (qRT-PCR) was used to assess expression of the germ cell markers, Mvh, Oct4, Rec8, Scp1, Scp3 and Stra8. Also, Oct4 and Mvh expressions at the protein level were assessed using immunocytochemistry; we evaluated alkaline phosphatase activity in addition to cell number and viability. Germ cell-specific marker expression was increased significantly in cells differentiated in KO-DMEM for both EB and monolayer protocols; the highest level was in cultures using the EB protocol. The highest cell proliferation rate was observed using the monolayer/KO-DMEM protocol and the lowest using the EB/DMEM protocol. Generally, KO-DMEM exhibited the greatest impact on germ cell differentiation and cell proliferation. Optimization of germ cell differentiation of ES cells requires careful selection of culture medium.

中文翻译：

---

培养基可影响小鼠胚胎干细胞的生殖细胞分化。

尽管胚胎干（ES）细胞可以分化为生殖细胞，但对于培养基对这一过程的影响知之甚少。我们调查了两种培养基对胚胎干细胞分化的能力的影响，使用胚胎样体（EB）和单层培养方案。在四种实验条件下，在小鼠ES细胞中诱导生殖细胞分化：EB / Dulbecco改良的Eagle培养基（EB / DMEM），EB /敲除Dulbecco改良的Eagle培养基（EB / KO-DMEM），单层/ Dulbecco改良的Eagle培养基（单层/ DMEM）和单层/剔除Dulbecco改良的Eagle培养基（单层/ KO-DMEM）。孵育6天后，使用定量实时聚合酶链反应（qRT-PCR）评估生殖细胞标记Mvh，Oct4，Rec8，Scp1，Scp3和Stra8。另外，使用免疫细胞化学评估了蛋白水平上的Oct4和Mvh表达。我们评估了碱性磷酸酶的活性以及细胞数量和活力。对于EB和单层实验方案，在KO-DMEM中分化的细胞中，生殖细胞特异性标志物的表达显着增加。最高水平是使用EB方案进行的培养。使用单层/ KO-DMEM方案观察到最高的细胞增殖速率，使用EB / DMEM方案观察到最低的细胞增殖速率。通常，KO-DMEM对生殖细胞分化和细胞增殖表现出最大的影响。ES细胞生殖细胞分化的优化需要仔细选择培养基。我们评估了碱性磷酸酶的活性以及细胞数量和活力。对于EB和单层实验方案，在KO-DMEM中分化的细胞中，生殖细胞特异性标志物的表达显着增加。最高水平是使用EB方案进行的培养。使用单层/ KO-DMEM方案观察到最高的细胞增殖速率，使用EB / DMEM方案观察到最低的细胞增殖速率。通常，KO-DMEM对生殖细胞分化和细胞增殖表现出最大的影响。ES细胞生殖细胞分化的优化需要仔细选择培养基。我们评估了碱性磷酸酶的活性以及细胞数量和活力。对于EB和单层实验方案，在KO-DMEM中分化的细胞中，生殖细胞特异性标志物的表达显着增加。最高水平是使用EB方案进行的培养。使用单层/ KO-DMEM方案观察到最高的细胞增殖速率，使用EB / DMEM方案观察到最低的细胞增殖速率。通常，KO-DMEM对生殖细胞分化和细胞增殖表现出最大的影响。ES细胞生殖细胞分化的优化需要仔细选择培养基。使用单层/ KO-DMEM方案观察到最高的细胞增殖速率，使用EB / DMEM方案观察到最低的细胞增殖速率。通常，KO-DMEM对生殖细胞分化和细胞增殖表现出最大的影响。ES细胞生殖细胞分化的优化需要仔细选择培养基。使用单层/ KO-DMEM方案观察到最高的细胞增殖速率，使用EB / DMEM方案观察到最低的细胞增殖速率。通常，KO-DMEM对生殖细胞分化和细胞增殖表现出最大的影响。ES细胞生殖细胞分化的优化需要仔细选择培养基。