重慶市華雅干細胞技術有限公司 ，  ：

華雅集團旗下授權代理TTAKARA Cellartis干細胞產品

DEF-CS是單個干細胞基因操作的理想選擇，和目前市場上的相關產品不同的是，DEF-CS在設計上就是支持單細胞生長的。

DEF-CS 500誘導多能干細胞無血清培養基

英文名稱：Cellartis® DEF-CS™ 500 Culture System

中文名稱：Cellartis® DEF-CS™ 500人iPS細胞無血清培養系統

特點：成分確定 無血清 無飼養層 *培養基

用途：iPS細胞大規模培養，單細胞培養，轉染和Scaffold接種。

保存：基礎培養基和包被劑2-8°C，添加劑-20°C

DEF-CS系統是一個強大的培養系統，用于在無飼養層和確定環境下高效擴增人誘導多能干細胞（iPS）。該系統能夠實現穩定的生長速率，適合傳統iPS細胞培養，細胞批量生產和單細胞培養。用此培養系統生長的細胞保持未分化狀態，不會有可檢測的背景分化，無需細胞選擇。酶法傳代的單細胞可用于單細胞相關應用，包括高通量篩選，轉染和支架接種（scaffold seeding）。

☆ 成分確定、無血清、無飼養層培養

☆ All-In-One型：培養液+添加劑+包被劑

☆ 細胞單層生長，可進行單細胞傳代

☆ 維持未分化狀態，無需篩選分化細胞

☆ 單個干細胞基因操作實驗的理想選擇

產品應用

·放大和大規模生產人類iPS誘導多能干細胞

·人類iPS誘導多能干細胞的單細胞培養

·轉染和重編程

·高通量篩選

·組織工程（支架接種）

DEF-CS是單個誘導多能干細胞基因操作的理想選擇

Human iPS和Human ES增殖用培養基(DEF-CS Culture System, all-in-one format)

作為一款創新型的產品，Cellartis DEF-CS培養系統使得單細胞干細胞操作成為常規的操作。該產品是專門針對人類誘導多能干細胞（hiPS）和人類胚胎干細胞（hES）研發的高效率增殖用培養基產品體系。該產品系統為all-in-one型，包括了所有組分，客戶不需要另外求購組分。該產品是成分確定的培養基產品，而且不需要飼養層。DEF-CS既可以實現單細胞培養，也可以用于傳統的iPS培養模式以及大規模干細胞增殖。在使用DEF-CS培養基增殖干細胞時，幾乎沒有背景分化的問題，這使得細胞篩選不再必需。作為一款創新產品，利用DEF-CS培養干細胞時，可以使用酶消化法（enzymatic passaging）實現需要單細胞操作，這一特點十分有利于高通量細胞鑒別篩選（high-throughput screening）、轉染（transfection）、支架接種（scaffold seeding）等。

Cellartis DEF-CS 500 Culture System 組份

培養系統包括基礎培養基、包被劑和添加劑

Cellartis DEF-CS 500 Culture System (Cat. No. Y30010)

500 ml Cellartis DEF-CS 500 Basal Medium (Cat. No. Y30011; not sold separay)

4 ml Cellartis DEF-CS 500 COAT-1 (Cat. No. Y30012)

Cellartis DEF-CS 500 Additives (Cat. No. Y30016)：

o 2 x 750 µl DEF-CS GF-1

o 500 µl DEF-CS GF-2

o 200 µl DEF-CS GF-3

需要自行準備試劑：PBS（含鈣鎂和不含鈣鎂），TrypLE選擇酶（無酚紅）

PBS Dulbecco's with Ca2+ & Mg2+ (D-PBS +/+)

PBS Dulbecco's w/o Ca2+ & Mg2+ (D-PBS –/–)

TrypLE Select Enzyme (1X), no phenol red

誘導多能干細胞培養性能

單細胞傳代

Human induced pluripotent stem (iPS) cells can be passaged as single cells in the Cellartis DEF-CS Culture System. A single GFP-actin iPS cell was isolated and placed in the well of a culture dish. Twenty-four hours after seeding, morphology was assessed by fluorescence microscopy at 20x (Panel A) and 40x (Panel B) magnification. Sixteen days later, the single GFP-actin iPS cell had proliferated into numerous cells as evidenced by microscopic observation at 4x (Panel C), 10x (Panel D), 20x (Panel E), and 40x (Panel F) magnification.

擴增潛力

Expansion potential of a characterized working bank of human induced pluripotent stem (iPS) cells in the Cellartis DEF-CS Culture System. The Cellartis DEF-CS Culture System can produce 2 x 109 human iPS cells within 4 passages (18–20 days) from frozen cells (2.0–2.5 x 106 cells).

多能干性維持

Human pluripotent stem cells remain undifferentiated when cultured in the Cellartis DEF-CS Culture System. Human iPS cells cultured for 23 passages in the Cellartis DEF-CS Culture System were characterized by Oct-4 staining (A) and nuclear staining (B).

參考文獻：

1. Sivertsson, Louise, et al. "Hepatic differentiation and maturation of human embryonic stem cells cultured in a perfused     three-dimensional bioreactor." Stem cells and development 22.4 (2012): 581-594.

2. Hanson, Charles, et al. "Transplantation of human embryonic stem cells onto a partially wounded human cornea in     vitro." Acta ophthalmologica 91.2 (2013): 127-130.

3. Norrman, Karin, et al. "Distinct gene expression signatures in human embryonic stem cells differentiated towards definitive endoderm at single-cell level." Methods 59.1 (2013): 59-70.

4. Ulvestad, Maria, et al. "Drug metabolizing enzyme and transporter protein profiles of hepatocytes derived from human embryonic and induced pluripotent stem cells." Biochemical pharmacology 86.5 (2013): 691-702.

5. Ramirez JM, et al. Side scatter intensity is highly heterogeneous in undifferentiated pluripotent stem cells and predicts clonogenic self-renewal. Stem Cells Dev.2013 Jun 15;22(12):1851-1860.

6. Borestrom, Cecilia, et al. “Footprint-free human induced pluripotent stem cells from articular cartilage with redifferentiation capacity: A first step toward a clinical-grade cell source.” Stem Cells Trans. Med. (2014) 3, 433-447.

7. Kia, Richard, et al. "MicroRNA-122: a novel hepatocyte-enriched in vitro marker of drug-induced cellular toxicity." Toxicological Sciences (2014): kfu269.

8. Valton, Julien, et al. "Efficient strategies for TALEN-mediated genome editing in mammalian cell lines." Methods 69.2 (2014): 151-170.

9. Zandén, Carl, et al. "Stem cell responses to plasma surface modified electrospun polyurethane scaffolds." Nanomedicine: Nanotechnology, Biology and Medicine 10.5 (2014): 949-958.

10. Asplund, Annika, et al. “One Standardized Differentiation Procedure Robustly Generates Homogenous Hepatocyte Cultures Displaying? Metabolic Diversity from a Large Panel of Human Pluripotent Stem Cells” Stem Cell Rev and Rep (2015)

 ?華雅集團旗下授權代理TTAKARA Cellartis干細胞產品，Cellartis的多能細胞培養系統包括iPS誘導多能干細胞株、iPS誘導多能干細胞和ES胚胎干細胞培養基，iPS基因編輯用單細胞克隆培養基和多能細胞檢測用單克隆抗體。

重慶市華雅干細胞技術有限公司以“專注干細胞研究，提升干細胞品質”為發展理念，專業從事干細胞基礎研究、干細胞再生醫學臨床應用研究及干細胞技術服務，自主研發產品熱賣中：人間充質干細胞無血清培養基 

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