StemPro™ hESC SFM - FAQs

人体ES细胞一般可通过以下方法进行鉴定：经典型的形态（它们以致密的细胞团形式生长 生长为紧密聚集的小型细胞团，细胞小并具有高核质比）；表面标志物的表达；干细胞特异性基因表达的RT-PCR检测（如Oct3/4，Sox2和Nanog）；碱性磷酸酶染色和端粒酶活性检测。最为常用的人ES细胞特异性表面标志物包括发育阶段特异性的胚胎抗原SSEA-3和SSEA-4。其它的ES细胞特异性的其它表面抗原还包括TRA-1-60和TRA-1-81。（Science 282:1145 (1998)）。

人胚胎干细胞源自人类囊胚内细胞团, 是通过使用兔抗血清检测BeWO细胞（人滋养层细胞系）抗血清的通过免疫外科法手段来进行分离的（Science 282:1145 (1998)）。

胚胎干（ES）细胞是源自早期哺乳动物胚胎的细胞，能够在体外进行无限的未分化增殖，同时保留分化为各类成体组织（包括生殖细胞）的潜能。ES细胞的多能性通常在体外通过以下方法来证明：将ES细胞诱导分化为类拟胚体，并以谱系特异性的标志物检测三个体胚层（内胚层，中胚层和外胚层）中分化细胞的谱系特异性标志物分化情况，或将这些细胞注射到免疫缺陷型小鼠中，以检测畸胎瘤中产生的细胞类型。

是的，这些产品就是在设计中就是配合使用的。

We recommend to thaw the supplement in the 37°C water bath, aliquot and freeze at -5°C to -20°C. Thaw each aliquot only one additional time. To make 100 ml complete medium, mix 90.8 ml DMEM/F-12 with GlutaMax matrix, 2 ml StemPRO hESC supplement, 7.2 ml 25% BSA solution, 80 ul bFGF (at 10 ug/ml), and 182 ul 2-mercaptoethanol (at 55mM). Store at 2°C-8°C until ready to use, but use the same day.

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No, we don't recommend StemPro hESC SFM for mouse cells because they showed poor growth and some differentiation. We recommend KnockOut DMEM and KnockOut SR for mouse ESC cultures (catalog numbers 10829-018 and 10828-028 respectively).

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Yes, derivations using StemPro hESC SFM have been carried out using either a basement membrane extract (like Geltrex growth factor) or feeder cells.

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StemPro hESC SFM has been tested with the following hESC: H1, H9, BG01, BG02, BG03, HUES6, HUES9, Cyth203, CyT49, BG01v, HES-2, HES-3, KhES-1, and TE06. The medium has also been used with the Rhesus monkey line R336.4. Some cells may require a short adaptation time but most do not.

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Yes, we recommend using Geltrex hESC-qualified Reduced Growth Factor basement membrane (catalog number A10480-02), CELLstart (catalog number A10142-01) or similar matrix.

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In addition to the StemPRO hESC SFM kit, you will also need the following to make a complete medium: full length recombinant human FGF (catalog # PHG0261) and 2-Mercaptoethanol (catalog # 21985-023).

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The StemPro hESC SFM kit (Cat. No. A1000701) contains: - 500 mL DMEM/F-12 with GlutaMax (Cat. No. 10565-018)
- 10 mL StemPro hESC SFM Supplement 50X (Cat. No. A10006-01)
- 40 mL Bovine Serum Albumin 25% (BSA Solution) (Cat. No. A10008-01)
Note that the StemPRO hESC Supplement and BSA solution cannot be ordered individually.

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Human ES cells are generally characterized by their typical morphology (they grow as tightly packed clusters of small cells with high ratio of nucleus to cytoplasm); surface marker expression; RT-PCR detection of stem cell-specific gene expression (such as Oct3/4, Sox2, and Nanog); alkaline phosphatase staining, and telomerase activity assay. The most commonly used ES specific surface markers include stage-specific embryonic antigens SSEA-3 and SSEA-4 for human ES cells. Other ES-specific surface antigens also include TRA-1-60 and TRA-1-81. (Science 282:1145 (1998).

Human ES cells are derived from human blastocyst inner cell masses, isolated by immunosurgery with rabbit antiserum to BeWO cells (a human trophoblast cell line) (Science 282:1145 (1998)).

Embryonic stem (ES) cells are derived from the early mammalian embryo and are capable of unlimited, undifferentiated proliferation in vitro while maintaining their potential to differentiate into a wide of range of adult tissues including germ cells. The pluripotency of the ES cells is normally demonstrated in vitro by inducing ES cells to differentiate into embryoid bodies and checking lineage-specific markers for differentiated cells in three body layers (endo, meso, and ectoderm), or injecting them into immunodeficient mice and determining the cell types produced in the teratomas.

Yes, these products were designed to work together.

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