US9782443B2
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Preparing tooth-like structure using stem cell
Assignee: Guangzhou Institutes of Biomedicine and Health
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Filing: 2013-02-05
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Granted: 2017-10-10
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Expires: 2033-02-05
Abstract
Abstract The usage of a stem cell in preparation of a tooth-like structure is provided. And a culture medium, a method for preparing an epithelial-like cell, a kit for preparing an ameloblast, a method for preparing an ameloblast are also provided. Specifically, the culture medium comprises a basal medium, which is DMEM/F12 medium; N2 supplement; retinoic acid; and BMP-4.
Inventors
- 👤 Duanqing Pei
- 👤 Jinglei CAI
- 👤 Pengfei Liu
- 👤 Shubin CHEN
- 👤 Yanmei Zhang
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-02 01:21
CPC Classifications
A; A61; A61K; A61K35/00; A61K35/12; A61K35/48; A61K35/54; A61K35/545; A61P; A61P1/00; A61P1/02; C; C07; C07K; C07K14/00; C07K14/435; C07K14/475; C07K14/51; C12; C12N; C12N5/00; C12N5/06; C12N5/0602; C12N5/0652; C12N5/0654; C12N2500/00; C12N2500/30; C12N2500/32; C12N2500/33; C12N2500/38; C12N2501/00; C12N2501/06; C12N2501/10; C12N2501/155; C12N2501/30; C12N2501/38; C12N2501/385; C12N2506/00; C12N2506/02; C12N2506/45
Claims Analysis
Total Claims: 18
Claim 1
Independent Claim
39 words
A method of preparing a tooth-like structure, wherein said tooth-like structure is prepared from a human induced pluripotent stem cell, and wherein said human induced pluripotent stem cell is prepared from a stem cell derived from a urine cell.
Claim 2
Independent Claim
39 words
A method of preparing a tooth-like structure, wherein said tooth-like structure is prepared from a human induced pluripotent stem cell, and wherein said human induced pluripotent stem cell is prepared from a stem cell derived from a urine cell.
Claim 3
Dependent Claim
21 words
The method of claim 1 , wherein the tooth-like structure is produced in a mammalian animal by means of xenograft transplantation.
Claim 4
Dependent Claim
26 words
The method of claim 2 , wherein the mammalian animal is at least one selected from a group consisting of mouse, rat, pig, dog, and monkey.
Claim 5
Dependent Claim
22 words
The method of claim 1 , wherein the stem cell is induced to form an epithelial-like cell prior to the xenograft transplantation.
Claim 6
Dependent Claim
13 words
The method of claim 1 , wherein the tooth-like structure comprises an ameloblast.
Claim 7
Independent Claim
120 words
A method for preparing an ameloblast, comprising: preparing an epithelial-like cell by a method comprising culturing a stem cell using a first culture medium comprising a DMEM/F12 medium supplemented with about 1 wt. % N2 supplement, about 1 μM retinoic acid and about 25 ng/ml BMP-4, to induce the stem cell differentiating into the epithelial-like cell; preparing a dental mesenchyme from a first animal; recombining the epithelial-like cell with the dental mesenchyme to obtain a recombinant specimen; culturing the resulting recombinant specimen to obtain a reconstituted explant; and transplanting the reconstituted explant into a second animal for at a predetermined time to form the ameloblast, wherein the stem cell is an induced pluripotent stem cell derived from a urine cell.
Claim 8
Independent Claim
120 words
A method for preparing an ameloblast, comprising: preparing an epithelial-like cell by a method comprising culturing a stem cell using a first culture medium comprising a DMEM/F12 medium supplemented with about 1 wt. % N2 supplement, about 1 μM retinoic acid and about 25 ng/ml BMP-4, to induce the stem cell differentiating into the epithelial-like cell; preparing a dental mesenchyme from a first animal; recombining the epithelial-like cell with the dental mesenchyme to obtain a recombinant specimen; culturing the resulting recombinant specimen to obtain a reconstituted explant; and transplanting the reconstituted explant into a second animal for at a predetermined time to form the ameloblast, wherein the stem cell is an induced pluripotent stem cell derived from a urine cell.
Claim 9
Dependent Claim
16 words
The method of claim 6 , wherein the stem cell is cultured for about 5-10 days.
Claim 10
Dependent Claim
29 words
The method of claim 6 , wherein the first animal and the second animal are independently selected from a group consisting of mouse, rat, pig, dog, and monkey respectively.
Claim 11
Dependent Claim
22 words
The method of claim 7 , wherein the first animal is a fetal mouse, and the second animal is a nude mouse.
Claim 12
Dependent Claim
30 words
The method of claim 9 , wherein the dental mesenchyme is mechanically separated from a tooth germ, and the tooth germ is digested using a protease prior to the separation.
Claim 13
Dependent Claim
16 words
The method of claim 10 , wherein the protease is a Dispase of about 0.75 mg/ml.
Claim 14
Dependent Claim
27 words
The method of claim 6 , wherein the resulting recombinant specimen is cultured using a DMEM medium supplemented with FBS, non-essential amino acid, glutamine, penicillin and streptomycin.
Claim 15
Dependent Claim
23 words
The method of claim 9 , wherein the recombinant tissue is transplanted into a space under a kidney capsule of the nude mouse.
Claim 16
Dependent Claim
16 words
The method of claim 6 , wherein the predetermined time is about 2 to 4 weeks.
Claim 17
Dependent Claim
14 words
The method of claim 6 , wherein the predetermined time is about 3 weeks.
Claim 18
Dependent Claim
93 words
The method of claim 6 , wherein the stem cell is cultured using at least one medium selected from the group consisting of mTeSR1 medium, E8 medium, and KSR-conditioned medium prior to preparing the epithelial-like cell, wherein the medium is replaced with the first culture medium when the cultured stem cell reach a confluence of about 60-80%, to obtain the epithelial-like cell, said first culture medium being DMEM/F12 medium supplemented with a N2 supplement of about 1 wt. %, a retinoic acid of about 1 μM, and a BMP-4 of about 25 ng/ml.
References Cited (Backward Citations)
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