Sunday, February 24, 2013

Non-colony type monolayer culture of human embryonic stem cells

Stem Cell Res. 2012 Nov;9(3):237-48. doi: 10.1016/j.scr.2012.06.003. Epub 2012 Jun 28.

Non-colony type monolayer culture of human embryonic stem cells.

Chen KG, Mallon BS, Hamilton RS, Kozhich OA, Park K, Hoeppner DJ, Robey PG, McKay RD.

Source

NIH Stem Cell Unit, National Institute of Neurological Disorders and Stroke, National Institutes of Health, Bethesda, MD 20892, USA. cheng@mail.nih.gov

Abstract

Regenerative medicine, relying on human embryonic stem cell (hESC) technology, opens promising new avenues for therapy of many severe diseases. However, this approach is restricted by limited production of the desired cells due to the refractory properties of hESC growth in vitro. It is further hindered by insufficient control of cellular stress, growth rates, and heterogeneous cellular states under current culture conditions. In this study, we report a novel cell culture method based on a non-colony type monolayer (NCM) growth. Human ESCs under NCM remain pluripotent as determined by teratoma assays and sustain the potential to differentiate into three germ layers. This NCM culture has been shown to homogenize cellular states, precisely control growth rates, significantly increase cell production, and enhance hESC recovery from cryopreservation without compromising chromosomal integrity. This culture system is simple, robust, scalable, and suitable for high-throughput screening and drug discovery.

参考文献　奇形腫

Methods Mol Biol. 2011;767:231-41. doi: 10.1007/978-1-61779-201-4_17.

The teratoma assay: an in vivo assessment of pluripotency.

Wesselschmidt RL.

Source

Center for Department of Applied Technology Development, Beckman Research Institute, City of Hope, Duarte, CA, USA. robin@primogenix.com

Abstract

A teratoma is a nonmalignant tumor comprised of a disorganized mixture of cells and small foci of tissue comprised of cells from all three of the embryonic germ-layers. By definition, a cell is pluripotent if it can differentiate into cells derived from all three of the embryonic germ-layers: ectoderm, mesoderm, and endoderm. In the teratoma assay, putative pluripotent stem cells (PSCs) are implanted into an immune-compromised mouse where they may proliferate and differentiate to form a teratoma. The PSCs grow at the implantation site supported by a complex mixture of factors from the local milieu, as well as circulating factors that are vital components of normal mammalian physiology. After a predetermined time of 6-12 weeks or when the tumor has reached sufficient size, it is removed and subjected to histopathological analysis. The teratoma may be further processed by immunocytochemistry and gene expression profiling. This chapter describes methods to generate teratomas through the implantation of putative PSC lines in the SCID mouse. Implantation at the following sites is described: (1) intramuscular, (2) subcutaneous, (3) under the testis capsule, and (4) under the kidney capsule.

Methods Mol Biol. 2011;690:67-80. doi: 10.1007/978-1-60761-962-8_4.

Functional assays for human embryonic stem cell pluripotency.

O'Connor MD, Kardel MD, Eaves CJ.

Source

Terry Fox Laboratory, BC Cancer Agency, Vancouver, BC, Canada.

Abstract

Realizing the potential that human embryonic stem cells (hESCs) hold, both for the advancement of biomedical science and the development of new treatments for many human disorders, will be greatly facilitated by the introduction of standardized methods for assessing and altering the biological properties of these cells. The 7-day in vitro alkaline phosphatase colony-forming cell (AP(+)-CFC) assay currently offers the most sensitive and specific method to quantify the frequency of undifferentiated cells present in a culture. In this regard, it is superior to any phenotypic assessment protocol. The AP(+)-CFC assay, thus, provides a valuable tool for monitoring the quality of hESC cultures, and also for evaluating quantitative changes in pluripotent cell numbers following manipulations that may affect the self-renewal and differentiation properties of the treated cells. Two other methods routinely used to evaluate hESC pluripotency involve either culturing the cells under conditions that promote the formation of nonadherent differentiating cell aggregates (termed embryoid bodies), or transplanting the cells into immunodeficient mice to obtain teratomas containing differentiated cells representative of endoderm, mesoderm, and ectoderm lineages.

PLoS One. 2012;7(9):e45532. doi: 10.1371/journal.pone.0045532. Epub 2012 Sep 25.

Standardization of the teratoma assay for analysis of pluripotency of human ES cells and biosafety of their differentiated progeny.

Gropp M, Shilo V, Vainer G, Gov M, Gil Y, Khaner H, Matzrafi L, Idelson M, Kopolovic J, Zak NB, Reubinoff BE.

Source

The Hadassah Human Embryonic Stem Cell Research Center, The Goldyne Savad Institute of Gene Therapy and Department of Gynecology, Hadassah-Hebrew University Medical Center, Jerusalem, Israel.

Nat Biotechnol. 2007 Nov;25(11):1212; discussion 1212.

The terminology of teratocarcinomas and teratomas.

Damjanov I, Andrews PW.

The terminology of teratocarcinomas and teratomas.

Lensch MW, Ince TA.

Nat Biotechnol. 2007 Nov;25(11):1211; author reply 1211-2.

An antibody against SSEA-5 glycan on human pluripotent stem cells enables removal of teratoma-forming cells.

Tang C, Lee AS, Volkmer JP, Sahoo D, Nag D, Mosley AR, Inlay MA, Ardehali R, Chavez SL, Pera RR, Behr B, Wu JC, Weissman IL, Drukker M.

Nat Biotechnol. 2011 Aug 14;29(9):829-34. doi: 10.1038/nbt.1947.

Nat Biotechnol. 2011 Sep 8;29(9):803-5. doi: 10.1038/nbt.1974.

Toward safer regenerative medicine.

Andrews PW.

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Lectin microarray論文が引用三位

Lectin microarray論文がGenes to Cellsに引用三位だそうで嬉しい。下の画像で、真ん中より下の方に遠慮がちに書かれています。Toyodaさん、よかったですね～

The Scientistにも今度記事になるとのことなので、紹介させて下さい。

Sunday, February 17, 2013

単語

Webよりコピペ。

Types of IND Applications

•	Commercial IND – submitted by a sponsor that intends to market the product upon FDA approval.

•	Investigator-Initiated IND – submitted by a physician who both initiates and conducts an investigation.

•	Emergency Use IND – issued by the FDA to allow the use of an experimental drug or biologic for the treatment of one patient when there are no other reasonable treatment options and there is not time for submission and review of a regular IND or for IRB review. Research may not be conducted under an emergency use IND. An emergency use IND exemption may be used one time only for a particular drug or biologic at a particular institution. Subsequent uses require prior CHR review and approval. For more information, please review the CHR Guidelines on the Emergency Use and Compassionate Use of Experimental Drugs and Devices.

•	Treatment Use of Investigational Drugs – submitted for experimental drugs showing promise in clinical testing for serious or life-threatening conditions while the final clinical work is conducted and the FDA review takes place. For more information, please see http://www.fda.gov/oc/ohrt/irbs/drugsbiologics.html-treatment. CHR review is required for any Treatment IND and Group C Treatment IND studies, even if the FDA has granted a waiver from local IRB requirements.