公开(公告)号：US20050266557A1

公开(公告)日：2005-12-01

申请号：US11113074

申请日：2005-04-25

申请人： Chantal Proulx ,  Nicolas Dupuis ,  Real Lemieux

发明人： Chantal Proulx ,  Nicolas Dupuis ,  Real Lemieux

IPC分类号： C12N5/02 ,  C12N5/078 ,  C12N5/0789 ,  C12N5/08

CPC分类号： C12N5/0647 ,  C12N5/0644 ,  C12N2500/02 ,  C12N2501/125 ,  C12N2501/145 ,  C12N2501/23 ,  C12N2501/26

摘要： Based on previous evidence suggesting positive effects of fever on in vivo hematopoiesis, the effect of hyperthermia on the expansion and differentiation of megakaryocytes (MKs) in ex vivo cultures of CB CD34-enriched cells has now been tested. Cells were cultured at 37° C. or 39° C. for 14 days in cytokine conditions optimized for MK development, and analyzed periodically by microscopy, flow cytometry and colony assays. Compared to 37° C., cultures maintained at 39° C. produced much more total cells (5X), MK progenitors (9X) and total MKs (7X), and showed accelerated (3-4 days) and enhanced MK maturation with increased yields of proplatelets and platelets (11.7X). The increased number of CD34+ cells and myeloid progenitors in the 39° C. cultures also suggested a general stimulatory effect of hyperthermia on the expansion of more primitive stem/progenitor cells and of cells of other lineages.

摘要翻译： 基于先前证据表明发热对体内造血作用的积极影响，现在已经测试了高热对CB CD34富集细胞离体培养物中巨核细胞（MK）的扩增和分化的影响。 细胞在37℃或39℃下在为MK发育优化的细胞因子条件下培养14天，并通过显微镜，流式细胞术和菌落测定定期分析。 与37℃相比，保持在39℃的培养物产生更多的总细胞（5X），MK祖细胞（9X）和总MK（7X），并且显示出加速（3-4天）和增强的MK成熟增加 产前血小板和血小板（11.7倍）。 在39℃培养物中CD34 +细胞和骨髓祖细胞数量的增加也表明热疗对更原始的干/祖细胞和其他谱系的细胞的扩增具有一般的刺激作用。

公开(公告)号：US5990385A

公开(公告)日：1999-11-23

申请号：US968688

申请日：1997-11-12

申请人： Louis-P. Vezina ,  Serge Laberge ,  Renee Bazin ,  Habib Khoudi ,  Real Lemieux ,  Guy Allard

发明人： Louis-P. Vezina ,  Serge Laberge ,  Renee Bazin ,  Habib Khoudi ,  Real Lemieux ,  Guy Allard

IPC分类号： C07K14/705 ,  C12N15/13 ,  C12N15/82 ,  A01H1/02 ,  C07K1/14 ,  C07K1/22

CPC分类号： C07K14/70503 ,  C12N15/8258

摘要： This invention is directed to characterizing a host system suitable for the production of functional transgenic proteins, such as anti-human IgG, for use in applications requiring Government regulatory approval. It is well known that regulatory agencies required stable, consistent master cell banks and master cell lines for the production of transgenic proteins in order to ensure sufficient material for appropriate characterization, clinical trials, and potential sales. Current plant production systems require the establishment of seed banks for this purpose. However, there are many draw backs related to such a system for the production of a continuous reliable transgenic protein source. An aspect of this invention is directed to characterizing a plant production system suitable for transgenic proteins that meet the stringent regulatory requirements. Another aspect of this invention exemplifies the production and characterization of an anti-human IgG for use as a blood grouping reagents, through the expression of corresponding genes in transgenic alfalfa plants. The cDNAs of the heavy and light chains of a human IgG-specific IgG2a(kappa) murine mAb (C5-1) were transferred into alfalfa through Agrobacterium infection. Transgenic plants expressing the light- and heavy-chain encoding mRNAs were obtained and plants from the F1 progeny (obtained by sexual crossing) were found to express fully assembled C5-1. Furthermore, the transgenic protein was stable in vivo, as well as during extraction and purification procedures. Purification yielded a unique H2L2 form with a reactivity indistinguishable from hybridoma-derived C5-1 in standardized serological tests. Results indicate that plant-derived transgenic proteins, such as mAbs can be used as diagnostic reagents as effectively as hybridoma-derived mAbs, and demonstrates the usefulness of the transformed alfalfa system to produce large amounts of proteins, including multimeric proteins such as mAbs.