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Polynucleated megakaryocytic cell and method for manufacturing platelets

A technology for megakaryocytes and platelets, applied in the field of platelet manufacturing, can solve the problem of non-muscle type II myosin that cannot be observed locally, and achieve the effects of promoting multinucleation, increasing the number, and shortening the time.

Active Publication Date: 2014-05-21
THE UNIV OF TOKYO
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Lodier et al. (Non-Patent Document 1) clarified that in the endomitosis of megakaryocytes, although a cleavage groove is formed, non-muscle type II myosin locally present in the contractile ring cannot be observed. Defect in extension

Method used

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  • Polynucleated megakaryocytic cell and method for manufacturing platelets
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  • Polynucleated megakaryocytic cell and method for manufacturing platelets

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Embodiment

[0171] 1. Preparation of Megakaryocytes Before Multinucleation

[0172] 1-1. Preparation of pre-multinucleated megakaryocytes from ES cells

[0173] In order to study the multinucleation of megakaryocytes, megakaryocytes before multinucleation were prepared from ES cells (see Patent Document 3 for details).

[0174] The human ES cell line [KhES-3] was cultured in the presence of 20ng / ml VEGF for 14 days to prepare a network structure, and the hematopoietic precursor cells removed from the network structure were recovered and planted on 10T1 / 2 cells to Reach cell number 1×10 5 / hole.

[0175] The hematopoietic precursor cells thus prepared were infected with pMx tet off c-MYC2A BMI1 retroviral vector integrating c-MYC-2A-BMI1 three times every 12 hours at MOI=10 (confirmed by Jurkat cells) to perform c-MYC and induction of BMI1 expression (Patent Document 3). The pMx tet off c-MYC2A BMI1 vector can express the c-MYC gene and BMI1 gene in the presence of estradiol, while...

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Abstract

An object of the present invention is to provide a method of promoting polynucleation of megakaryocytic cells to form polynucleated megakaryocytic cells with higher polynucleation, and to further provide a method for efficiently producing platelets from polynucleated megakaryocytic cells. The present invention provides a method for producing polynucleated megakaryocytic cells, wherein the method includes a step for inducing forced expression of an apoptosis suppressor gene in megakaryocytic cells prior to polynucleation and culturing of these cells.

Description

technical field [0001] The present invention relates to a method for efficiently multinucleating pre-multinucleated megakaryocytes, a method for producing platelets from the megakaryocytes, and the like. Background technique [0002] In the treatment or surgical treatment of blood-related diseases, more blood cells are required. Among blood cells, platelets are one of the most important blood cells as cells necessary for blood coagulation and hemostasis. Platelets are required in large quantities for treatments such as leukemia, bone marrow transplantation, and anticancer therapy, and the need for a stable supply is high. Conventionally, in order to ensure the supply of platelets, in addition to the method of extracting blood from blood donors, the method of administering preparations such as TPO-like analogs (mimetics), the method of differentiating megakaryocytes from umbilical cord blood or bone marrow cells, etc. have been used. In recent years, techniques for inducing...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/10A61K35/14A61P7/00C12N15/09A61K35/19
CPCC12N2501/48A61K35/19C12N2501/165C12N2506/02C12N5/0644A61P7/00C12N5/0634A61K35/14C12N5/10C12N15/85
Inventor 江藤浩之中内启光高山直也中村壮
Owner THE UNIV OF TOKYO
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