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Method for inducing differentiation of umbilical cord mesenchymal stem cells toward renal tubular epithelial cells

A technology of mesenchymal stem cells and epithelial cells, applied in the biological field, can solve problems such as undifferentiated methods, and achieve the effect of increasing the induction rate

Active Publication Date: 2019-06-07
上海葆年生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there is no efficient method for inducing umbilical cord-derived mesenchymal stem cells to differentiate into renal tubular epithelial cells

Method used

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  • Method for inducing differentiation of umbilical cord mesenchymal stem cells toward renal tubular epithelial cells
  • Method for inducing differentiation of umbilical cord mesenchymal stem cells toward renal tubular epithelial cells
  • Method for inducing differentiation of umbilical cord mesenchymal stem cells toward renal tubular epithelial cells

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] 1 material

[0021] Timed pregnant mice were from Shanghai Slack Experimental Animal Co., Ltd.

[0022] The human umbilical cord samples were obtained from the hospital, and the parturients and their family members gave informed consent to the experiment, which was approved by the hospital ethics committee.

[0023] LG-DMEM basal medium was purchased from Hyclone Company, fetal bovine serum (FBS), EGF, type II collagenase, hyaluronidase and DNase were purchased from Gibco Company, all-trans retinoic acid (ATRA), activin, penicillin, Streptomycin and astragaloside IV were purchased from Sigma.

[0024] 2 methods

[0025] 2.1 Isolation and Characterization of Embryonic Kidney Tissue and Preparation of Conditioned Medium

[0026] The day of gestation of mice was determined according to the appearance of vaginal plug. On the 13th day of gestation, the pregnant mice were dissected, and the metanephros of the embryos were isolated, and the DMEM medium (Gibco, Gaithersburg...

Embodiment 2

[0058] During the research and development process, the inventor of the present application found that the existing commercially available centrifuge tubes cannot effectively meet the centrifugation requirements during use. For example, the existing commercially available centrifuge tubes often adopt a split structure. The centrifuge tube is connected with the tube cover through thread rotation. If the tube cover is lost or forgotten, the centrifuge tube cannot be used. Second, after the centrifuge tube is placed in the centrifuge, due to the difference in size, it is easy to shake in the placement tank and cannot be well fixed in the centrifuge. The centrifuge tube is easily damaged in the centrifuge's storage tank. Therefore, the applicant also made corresponding innovations to the centrifuge tube during the test, so that the centrifugation step meets the requirements of this application.

[0059] see Figure 1 to Figure 3, the improved centrifuge tube includes a centrifuge ...

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PUM

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Abstract

The invention discloses a method for inducing differentiation of umbilical cord mesenchymal stem cells toward renal tubular epithelial cells. The method comprises the following steps: culturing umbilical cord mesenchymal stem cells by an induction medium, wherein the induction medium is an LG-DMEM medium and contains FBS with final volume fraction of (8-12)%, RA with final concentration of 10<-8>-10<-6> M, EGF with final concentration of 9-11 ng / mL, penicillin with final concentration of 90-110 U / mL, streptomycin with final cocnnetration of 90-110 U / mL and a UB cell condition medium with finalvolume fraction of (8-12)%. Efficient induction of differentiation of umbilical cord mesenchymal stem cells toward renal tubular epithelial cells in vitro is realized, and cell donors are provided for cell therapy of kidney diseases.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a method for inducing differentiation of umbilical cord mesenchymal stem cells into renal tubular epithelial cells. Background technique [0002] Drug-induced renal injury manifests as acute tubular injury, and even tubular necrosis, leading to renal failure. The method for treating renal failure usually adopts dialysis and kidney transplantation. Due to the deficiency of kidney source and immune rejection, the therapeutic effect is not good. With the in-depth study of stem cells, stem cells have become the preferred source of donors for cell therapy. At present, significant progress has been made in the research on inducing the directed differentiation of stem cells. For example, CN109022361A discloses that CD34+CD126- cells from umbilical cord blood are separated and screened, and co-cultured with umbilical cord mesenchymal stem cells to induce differentiation of megakaryotic prog...

Claims

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Application Information

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IPC IPC(8): C12N5/071
Inventor 邓炎雷
Owner 上海葆年生物科技有限公司
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