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Method for producing osteoblast cluster using ips cells

An osteoblast, construct technology, applied in biochemical equipment and methods, animal cells, vertebrate cells, etc., can solve the lack of extracellular matrix, can not always provide a good prognosis, osteoinductive activity is lower than autologous bone, etc. problem, to achieve the effect of high bone regeneration ability

Pending Publication Date: 2021-11-30
TOHOKU UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] Nonabsorbable materials (such as hydroxyapatite) and absorbent materials (such as β-tricalcium phosphate) currently clinically used as artificial bone / bone substitute materials have, for example, lower osteoinductive activity than autologous bone and cannot be used in surgical interventions. Questions that always provide a good prognosis
In addition, hybrid artificial bone / bone substitute materials obtained by combining artificial bone with growth factor proteins such as bone morphogenetic protein (BMP) (the next-generation type requiring further development) lack "extracellular matrix" important for bone tissue regeneration , and therefore cannot provide sufficient bone regeneration

Method used

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  • Method for producing osteoblast cluster using ips cells
  • Method for producing osteoblast cluster using ips cells
  • Method for producing osteoblast cluster using ips cells

Examples

Experimental program
Comparison scheme
Effect test

Embodiment

[0171] Experiment 1: Screening of the optimal microspace size for the induction of iPS cells into osteoblast constructs

[0172] "method"

[0173] 1.1. Culture of mouse iPS cells

[0174] For this experiment, an iPS cell line established from mouse gingival fibroblasts was used [PLoS ONE, 5(9):e12743, 2010]. By using ES medium [containing 15% fetal bovine serum (Gibco / Life Technologies, GrandIsland, NY, USA), 2 mM L-glutamine (Wako Pure Chemical, Osaka), 1×10 -4 M non-essential amino acids (Life Technologies, Grand Island, NY, USA), 1×10 -4 M 2-Mercaptoethanol (Life Technologies, Grand Island, NY, USA), 50 U penicillin and 50 μg / ml streptomycin (Wako Pure Chemical) in Dulbecco's modified Eagle's medium (DMEM: containing 4.5 g / L glucose and no pyruvate Na; Nacalai Tesque, Kyoto)] These mouse iPS cells were maintained and cultured as iPS cell constructs on SNLP76.7-4 feeder cells (mitomycin C treated).

[0175] 1.2. Generation of osteoblast constructs of mouse iPS ce...

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Abstract

The invention provides a method for producing an osteoblast cluster from iPS cells, said method comprising: (1) a step for inducing the formation of embryoid bodies by non-adhesively culturing undifferentiated iPS cells; (2) a step for inducing the differentiation of the iPS cells into mesodermal cells by non-adhesively culturing the embryoid bodies of the iPS cells obtained in step (1); and (3) a step for inducing the differentiation into osteoblasts by non-adhesively culturing the mesodermal cells of the iPS cells obtained in step (2), wherein the aforesaid steps (1) and (2) are carried out using a culture container which is provided with a bottom face and a circular side wall standing from the bottom face and in which a plurality of concave portions are formed in the bottom face in such a manner as to be independent from each other.

Description

technical field [0001] [Cross-reference to related patent applications] [0002] This application claims the benefit of priority from Japanese Patent Application No. 2019-033277 filed on February 26, 2019, the entire contents of which are incorporated herein by reference. The present invention relates to a method for producing osteoblast constructs using iPS cells. Background technique [0003] There is an extremely high demand for artificial bone / bone replacement materials to replace defective sites such as bone loss due to bone tumor resection, comminuted fractures, bone defects associated with rheumatoid arthritis fixation, alveolar ridge resorption, etc. . [0004] Non-absorbable materials (such as hydroxyapatite) and resorbable materials (such as β-tricalcium phosphate) currently clinically used as artificial bone / bone replacement materials have, for example, lower osteoinductive activity than autologous bone and cannot be used in surgical interventions. Questions th...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/077C12N5/10A61L27/38
CPCA61L2430/02A61L27/3821A61L27/3847A61L27/3895A61L27/3834C12N2506/45C12N5/0654C12N2535/00C12N5/0696C12N2500/30
Inventor 江草宏大川博子堀江尚弘近藤威
Owner TOHOKU UNIV
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