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KR-102960920-B1 - COMPOSITION FOR MAINTAING DIFFERENTIATION POTENCY OF DIFFERENTIATED FIBROBLAST-LIKE CELLS

KR102960920B1KR 102960920 B1KR102960920 B1KR 102960920B1KR-102960920-B1

Abstract

By using the culture medium composition according to the present invention, not only can it contribute to the improvement of differentiation ability, efficacy, and stability of differentiated cells, but it can also increase cell growth and viability. Through this, the cultured cells become more stabilized, allowing for higher expression of active ingredients with each passage. Furthermore, the method for maintaining stable culture according to the present invention can extend the culture period compared to conventional culture methods, thereby extending the usable cell count and enabling efficient mass production at low cost. The differentiated fibroblast-like cells and culture medium maintained as described above can provide cosmetic compositions with skin improvement or excellent anti-aging effects, and furthermore, can be utilized in various industries, such as cell therapy agents for treating skin wrinkles or scars.

Inventors

  • 용은지

Assignees

  • 주식회사 래디안

Dates

Publication Date
20260507
Application Date
20221128

Claims (14)

  1. A composition for maintaining the differentiation ability of differentiated fibroblast-like cells differentiated from human adipose-derived mesenchymal stem cells, comprising DMEM/HIGH GLUCOSE medium containing transforming growth factor (TGF-β), fibroblast growth factor (FGF), dexamethasone, connective tissue growth factor (CTGF), insulin growth factor (IGF), epidermal growth factor (EGF), fetal bovine serum, penicillin, and streptomycin.
  2. delete
  3. A composition for maintaining the differentiation ability of differentiated fibroblast-like cells according to claim 1, characterized in that the differentiated fibroblast-like cells are differentiated from human adipose-derived mesenchymal stem cells by culturing human adipose-derived mesenchymal stem cells on a gelatin-coated plate and adding a replaceable cell differentiation medium, replacing it at intervals of 2 to 4 days and 2 to 7 times.
  4. A composition for maintaining the differentiation ability of differentiated fibroblast-like cells according to claim 3, wherein the cell differentiation medium is a DMEM/HIGH GLUCOSE medium comprising transforming growth factor (TGF-β), fibroblast growth factor (FGF), dexamethasone, connective tissue growth factor (CTGF), insulin growth factor (IGF), epidermal growth factor (EGF), fetal bovine serum, penicillin, and streptomycin.
  5. A method for culturing differentiated fibroblast-like cells differentiated from human adipose-derived mesenchymal stem cells using a composition that maintains differentiation ability according to claim 1.
  6. A culture medium containing differentiated fibroblast-like cells cultured according to the method of claim 5.
  7. In claim 6, the culture medium containing differentiated fibroblast-like cells is characterized by increasing the expression of one or more of Fibronectin and VEGF.
  8. In claim 6, the culture medium containing differentiated fibroblast-like cells is characterized in that the differentiated fibroblast-like cells or the culture medium thereof have a skin improvement or anti-aging effect.
  9. A culture medium containing differentiated fibroblast-like cells according to claim 6, characterized in that the differentiated fibroblast-like cells or the culture medium thereof are included in either a pharmaceutical composition or a cosmetic composition.
  10. A culture medium containing differentiated fibroblast-like cells according to claim 6, characterized in that the differentiated fibroblast-like cells or the culture medium thereof are included in any one of a pack, a band, or a patch.
  11. A culture medium containing differentiated fibroblast-like cells according to claim 10, characterized in that the pack is one of a local patch, sheet type, gauze type, cream type, and gel type.
  12. A culture medium containing differentiated fibroblast-like cells according to claim 10, characterized in that the pack is one selected from a mask pack, a hand pack, a foot pack, a calf pack, a hair pack, a thigh pack, an abdomen pack, a side pack, an upper arm pack, a chin pack, and a neck pack.
  13. A culture medium containing differentiated fibroblast-like cells according to claim 6, characterized in that the differentiated fibroblast-like cells or the culture medium thereof are in the form of any one of aerosol, liquid, solution, gel, paste, cream, ointment, foam, and powder.
  14. A cell therapy agent for treating skin wrinkles or scars, comprising the differentiated fibroblast-like cells of claim 6.

Description

Composition for maintaining differentiation potential of differentiated fibroblast-like cells The present invention relates to a method for maintaining the differentiated state of human adipose tissue-derived mesenchymal stem cells after differentiation into fibroblast-like cells, and a culture medium composition thereof. Stem cells refer to undifferentiated cells capable of differentiating into various types of cells that constitute biological tissues under specific environmental conditions. Due to these unique characteristics, stem cells are attracting attention in fields such as cell therapy, new drug development, and cosmeceuticals; recently, numerous new technologies, cell therapies, and cosmetic ingredients utilizing stem cells are being launched. In particular, while the plasticity to differentiate into various cell types is the greatest advantage of stem cells, the differentiation process is often prolonged due to various factors, or the differentiated cells frequently fail to produce sufficient effects. Furthermore, this differentiation capability has raised concerns regarding the stability of stem cells, leading to a high risk of cancer formation. Therefore, there are challenges that must be addressed before cells induced to differentiate from stem cells into specific cell lineages can be utilized. To realize the potential for the commercialization of stem cells, it is crucial to obtain large quantities of cells through cell culture. However, as the culture period lengthens, stem cells inevitably undergo aging and deformation. This leads to a decrease in stem cell capabilities, such as proliferative or differentiation ability, resulting in a problem where the sustainability of their effects is reduced. Furthermore, since prolonged culture periods lead to higher production costs, including material and labor expenses, there is a growing need to address these issues. As prior art, Patent Document 1 discloses a method for differentiating fibroblast-like cells from adipose-derived stem cells and a culture medium composition, but there is no recognition or disclosure of a composition for maintaining differentiation ability. Figure 1 shows the results of confirming the differentiation markers of early fibroblast-like cells obtained from Example 1. Figure 2 shows the results of confirming differentiation markers by passage measured according to Experimental Example 2. Figure 3 shows the results of confirming Fibronectin expression ability by passage measured according to Experimental Example 3. Figure 4 shows the results of confirming VEGF expression ability by passage measured according to Experimental Example 3. Hereinafter, embodiments of the present invention are described in detail so that those skilled in the art to which the present invention pertains can easily implement it. The embodiments of the present invention are provided to more completely explain the present invention to those with average knowledge in the art. Accordingly, the embodiments of the present invention may be modified in various different forms, and the scope of the present invention is not limited to the embodiments described below. Throughout the specification of the present invention, when a part is described as “comprising” a certain component, this means that, unless specifically stated otherwise, it does not exclude other components but includes additional components. Throughout the specification of the present invention, when a step is described as being located “on” or “before” another step, this may include not only cases where a step is in a direct chronological relationship with another step, but also cases where there is an indirect chronological relationship in which the chronological order of the two steps may change, such as a mixing step following each step. Throughout the specification of this invention, terms such as “about,” “substantially,” etc., are used to mean at or near the stated value when inherent manufacturing and material tolerances are presented in the said meaning, and are used to prevent unscrupulous infringers from unfairly exploiting the disclosure in which precise or absolute values are mentioned to aid in understanding the invention. Throughout the specification, the terms “step” or “step of” do not mean “step for”. The method for differentiating human adipose-derived mesenchymal stem cells into fibroblasts according to the present invention comprises culturing human adipose-derived mesenchymal stem cells on a gelatin-coated plate and adding a replaceable cell differentiation medium to differentiate them into fibroblasts. The plate according to the present invention must use an appropriate extracellular matrix molecule for the differentiation and proliferation of stem cells. Suitable extracellular matrix molecules include collagen, fibronectin, matrigel, and gelatin. In the present invention, gelatin, which exhibited the highest differentiation and proliferation rates, was preferably used, a