METHOD FOR PREPARING BONE MARROW CELL AGGREGATE
|Posted date||Sep 23, 2016|
|International application number||2016JP051009|
|International publication number||WO 2016121512|
|Date of international filing||Jan 14, 2016|
|Date of international publication||Aug 4, 2016|
|Title||METHOD FOR PREPARING BONE MARROW CELL AGGREGATE|
|Abstract||Provided is a technique whereby bone marrow cells can be organized by a simple method within a short period of time. A method for preparing a bone marrow cell aggregate, said method comprising adding a bone marrow cell mass-containing liquid to a medium that contains a swellable material, and culturing the bone marrow cell mass in the presence of the swellable material. A method for reconstructing a bone marrow tissue, said method comprising adding a bone marrow cell mass-containing liquid to a medium that contains a swellable material, and culturing the bone marrow cell mass in the presence of the swellable material. As the common sense in the art, it has been believed that, after disintegrating a bone marrow tissue, cell masses can be hardly reorganized while keeping the cell composition unchanged, i.e., without adding "a binder" such as adhesive cells or intracellular matrix. In fact, such cell masses cannot be aggregated by methods generally employed in the art. The present invention provides a remarkable breakthrough technique relating to the three-dimensional culture of bone marrow cells to thereby overcome the above problem and situation encountered in the prior art. It is also confirmed that a bone marrow-like tissue reconstructed by the method according to the present invention can be continuously cultured in a methyl celluose-containing medium until the 14th day.|
|Outline of related art and contending technology||
Such as a mouse or human bone marrow tissue is, play an important role as a hematopoietic tissue. Hematopoietic and clarification of the mechanism for disease related thereto, the bone marrow tissue reconstruction technique plays an important role. However, for most blood cells in bone marrow Mn, fall apart once again and it is difficult to organize.
As the bone marrow for culture techniques, known Dexter culture (non-patent document 1: Dexter TM, Allen TD, Lajtha LG. Conditions controlling the proliferation of haemopoietic stem cells in vitro. J Cell Physiol. 1977 Jun; 91 (3): 335-44.). This is because the stromal cells as a feeder, on which the maintenance of hematopoietic stem cells, to differentiate into a variety of cell culture method allows. Planar culture is performed in that environment.
The culture of 3 using a technique of performing a three-dimensional scaffold present (non-patent document 2: Nichols JE, Cortiella J, Lee J, Niles JA, Cuddihy M, Wang S, Bielitzki J, Cantu A, Mlcak R, Valdivia E, Yancy R, McClure ML, Kotov NA. In vitro analog of human bone marrow from 3D scaffolds with biomimetic inverted colloidal crystal geometry. Biomaterials. 2009 Feb; 30 (6): 1071-9, non-patent document 3: Leisten I, Kramann R, Ventura Ferreira MS, Bovi M, Neuss S, Ziegler P, Wagner W, Knuchel R, Schneider RK. 3D co-culture of hematopoietic stem and progenitor cells and mesenchymal stem cells in collagen scaffolds as a model of the hematopoietic niche. Biomaterials. 2012 Feb; 33 (6): 1736-47.).
Sawa Harvard University Wyss Institute birds 3 - dimensional reconstruction of the bone marrow via the courage et al. for utilizing the microfluidic device by the method reported study (non-patent document 4: Yu-suke Torisawa, Catherine S Spina, Tadanori Mammoto, Akiko Mammoto, James C Weaver, Tracy Tat, James J Collins, Donald E Ingber, Nature Methods, Vol11, JUNE, 663-669 2014) Bone marrow-on-a-chip replicates hematopoietic niche physiology in vitro,. They can be confirmed with the various hematopoietic phenomenon and in vitro. The actual reconstruction procedures include, cylindrical poly(dimethylsiloxane) (PDMS) in the hollow portion of bone to reconstruct the packed material, such as a mouse can be implanted into the body, to reconstruct the bone and bone marrow. Is embedded at the device body, not at all using live cells, bone marrow cells may be blood to move toward the via. The duration of implantation is required for 8 weeks. 8 Weeks after the bone marrow in the body tissue and the tissue was removed, the microfluidic device is used in in vitro perfusion hematopoietic phenomenon can be examined.
|Scope of claims||
|IPC(International Patent Classification)|
National States: AE AG AL AM AO AT AU AZ BA BB BG BH BN BR BW BY BZ CA CH CL CN CO CR CU CZ DE DK DM DO DZ EC EE EG ES FI GB GD GE GH GM GT HN HR HU ID IL IN IR IS JP KE KG KN KP KR KZ LA LC LK LR LS LU LY MA MD ME MG MK MN MW MX MY MZ NA NG NI NO NZ OM PA PE PG PH PL PT QA RO RS RU RW SA SC SD SE SG SK SL SM ST SV SY TH TJ TM TN TR TT TZ UA UG US UZ VC VN ZA ZM ZW
ARIPO: BW GH GM KE LR LS MW MZ NA RW SD SL SZ TZ UG ZM ZW
EAPO: AM AZ BY KG KZ RU TJ TM
EPO: AL AT BE BG CH CY CZ DE DK EE ES FI FR GB GR HR HU IE IS IT LT LU LV MC MK MT NL NO PL PT RO RS SE SI SK SM TR
OAPI: BF BJ CF CG CI CM GA GN GQ GW KM ML MR NE SN ST TD TG
Contact Information for " METHOD FOR PREPARING BONE MARROW CELL AGGREGATE "
- Public University Corporation Yokohama City University Research Promotion Division
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