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Culture medium and application thereof

A technology of culture medium and conditional medium, which is applied in the field of biomedicine and can solve the problems that the preparation method needs to be improved.

Active Publication Date: 2015-03-11
GUANGZHOU INST OF BIOMEDICINE & HEALTH CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] However, the preparation methods of odontogenic cells, especially ameloblasts, which are important for tooth regeneration in oral regenerative medicine, still need to be improved.

Method used

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  • Culture medium and application thereof
  • Culture medium and application thereof
  • Culture medium and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] Example 1: Preparation of epithelioid cells

[0044] Material: Induced pluripotent stem cells (UC-iPSC) derived from urine cells

[0045] Medium: mTeSR1 TM Medium (STEMCELL catalog#05850); DMEM / F12 culture supplemented with N2supplement (Invitrogen catalog#17502-048), retinoic acid (Sigma catalog#R2625) and BMP-4 (R&D catalog#314-BP-050 / CF) Base (Gibco catalog#11330-032), wherein the medium contains 1% by weight of N2supplement, 1 μM retinoic acid, and 25ng / ml BMP-4.

[0046] Method steps:

[0047] First, use Metrigel to coat the culture plate, then inoculate induced pluripotent stem cells on the culture plate, and use mTeSR1 TM The medium is used to cultivate it, and it is subcultured every 4-6 days. Among them, before passaging, the cultured H1ESC and UC iPSC should be digested into small pieces with 2 mg / ml Dispase enzyme.

[0048] When the induced pluripotent stem cells grow to 60%-70% of the bottom of the dish, add DMEM / F12 medium supplemented with N2supplemen...

Embodiment 2

[0050] Example 2: Preparation of ameloblasts

[0051] Materials: epithelial-like cells obtained in Example 1; molar tooth germs of 14.5-day ICR fetal mice; nude mice

[0052] Reagents: Dispase (Gibco catalog#17105-041) at a concentration of 0.75mg / ml; supplemented with FBS (PAA catalog#A11-151), glutamine (Gibco catalog#25030-081), non-essential amino acids (Gibco catalog# 11140-050) and penicillin (Hyclone catalog#SV30010) high glucose DMEM medium (Hyclone catalog#SH30022.01B), wherein, the medium contains 10% FBS, 2mM glutamine, 0.1mM non-essential amino acids , 100U / ml penicillin, 0.1mg / ml streptomycin.

[0053] Instruments: stereo microscope (ZEISS, SteReo Lumar V12); upright biological microscope ZEISS (Axio Scope A1)

[0054] Method steps:

[0055] 1. Obtaining mesenchyme from molar tooth germs of ICR fetal rats

[0056] The molar tooth germs of 14.5-day-old ICR fetal mice were mechanically isolated. Specifically, after digesting the tooth germ with Dispase at a conce...

Embodiment 3

[0065] According to the method for preparing ameloblasts described in Example 2, 8 different induced pluripotent stem cell clones derived from urine cells were used to prepare ameloblasts, and the efficiencies of each stem cell induced and prepared to obtain ameloblasts were calculated and compared. The results See Table 1 below:

[0066] Table 1

[0067] induced pluripotent stem cell clone

Number of teeth / number of reconstituted tissue samples

Tooth ratio

vUC1-iPSC-C1

6 / 42

14.3％

UC1-iPSC-C1

12 / 50

24％

UC5-iPSC-C1

13 / 56

23.2％

UC5-iPSC-C2

15 / 59

25.4％

UC5-iPSC-C3

6 / 40

15％

UC7-iPSC-C3

12 / 52

23.1％

UC7-iPSC-C6

7 / 60

11.7％

UC7-iPSC-C9

17 / 59

28.8％

[0068] Among them, among the 8 different induced pluripotent stem cell clones, vUC1-iPSC-C1 is an induced pluripotent stem cell clone obtained by retrovirus induction, and the other 7 strains are in...

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Abstract

The invention discloses a culture medium and application thereof. The culture medium is a DMEM / F12 culture medium containing N2 supplement, retinoic acid and BMP-4. With the culture medium provided by the invention, stem cells can be effectively induced into epithelioid cells; then, the obtained epithelioid cells and mesenchyme of animal dental germs are subjected to recombination, culture and transplantation, so adamantoblast in a dental sample structure can be effectively obtained.

Description

technical field [0001] The invention relates to the field of biomedicine, in particular to a culture medium and its application, more specifically, to a culture medium, a method for preparing epithelioid cells, a kit for preparing ameloblasts and a method for preparing ameloblasts. Background technique [0002] At present, inducing stem cells into specific tissue cell types and using them in the treatment of corresponding tissue lesions is a new way to treat many diseases at this stage. In recent years, researchers in the field of regenerative medicine and tissue engineering have mainly used stem cells as seed cells to induce the preparation of specific tissue cell types, thereby providing a large number of materials for clinical transplantation of cells, tissues or organs, and then used in Diabetes, Parkinson's disease, spinal cord injury, leukemia, myocardial injury, renal failure, liver cirrhosis and other diseases, and great progress has been made so far. [0003] Howev...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/0735C12N5/071C12N5/074
Inventor 裴端卿蔡景蕾刘朋飞陈树彬张燕梅
Owner GUANGZHOU INST OF BIOMEDICINE & HEALTH CHINESE ACAD OF SCI
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