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www.lonza.com U.S. Scientific Support: 800-521-0390 [email protected] EU/ROW Scientific Support: +49-221-99199-400 [email protected] Document # TS-CC-323-4 06/11 © 2011 Lonza Walkersville, Inc. Poietics™ hematopoietic progenitor growth medium HPGM™ Erythroid progenitors PT-3926 + Introduction HPGM™-hematopoietic progenitor growth medium (catalog number PT-3926) is a serum-free medium containing only human proteins that supports the proliferation of human hematopoietic progenitor cells + + (CD34 cells). CD34 cells can be stimulated to undergo a limited amount of proliferation and still maintain a relatively undifferentiated phenotype when grown in this medium. Additionally, HPGM™ can be used as the growth medium to support the differentiation of hematopoietic progenitor cells, such as erythroid progenitors, shown below. CD36 cells (2C-250 or 2G-250) should be 5 established in culture at a density of 2 x 10 to 6 1 x 10 cells per ml of HPGM™ supplemented with EPO (3 U/ml), SCF (25 ng/ml), IL-3 (10 ng/ml), and IL-6 (10 ng/ml). After 5 to 7 days in culture in HPGM™, glycophorin A and hemoglobin expression will be observed in greater than 80% of the cells. A small portion of the population can be expected to continue to mature, will express glycophorin A in the absence of CD71 expression, and will actually enucleate. Hematopoietic progenitors + CD34 cells should be established in culture at a 4 5 density of 2 x 10 to 2 x 10 cells per ml of HPGM™ and supplemented with cytokines - e.g. SCF (25 ng/ml), TPO (50 ng/ml), and FL (50 ng/ml). After 7 days in culture in HPGM™ the total cell population + should expand by 10 to 20-fold and the CD34 cells should expand by 5 to 8-fold. Other cytokine combinations can be used with varying results. + CD34 cells cultured in HPGM™ All trademarks herein are marks of Lonza Group or its subsidiaries. + CD36 cells cultured in HPGM™ When placing an order or for scientific support, please refer to the product numbers and descriptions listed above. For a complete listing of all Poietics™ products, refer to the Lonza website or our current catalog. To obtain a catalog, additional information or scientific support you may contact Lonza by web, e-mail, telephone, fax or mail. THESE PRODUCTS ARE FOR RESEARCH USE ONLY. Not approved for human or veterinary use, for application to humans or animals, or for use in clinical or in vitro procedures. WARNING: CLONETICS™ AND POIETICS™ PRODUCTS CONTAIN HUMAN SOURCE MATERIAL, TREAT AS POTENTIALLY INFECTIOUS. Each donor is tested and found non-reactive by an FDA approved method for the presence of HIV-I, hepatitis B virus and hepatitis C virus. Where donor testing is not possible, cell products are tested for the presence of viral nucleic acid from HIV, hepatitis B virus, and hepatitis C virus. Testing can not offer complete assurance that HIV-1, hepatitis B virus, and hepatitis C virus are absent. All human sourced products should be handled at the biological safety level 2 to minimize exposure of potentially infectious products, as recommended in the CDC-NIH manual, Biosafety in Microbiological and Biomedical Laboratories, 5th edition. If you require further information, please contact your site safety officer or scientific support. All trademarks herein are marks of Lonza Group or its subsidiaries.