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Research on stem cells isolated and cultured human umbilical cord charging method

Author: LiZuo
Tutor: ZhangYuGuang
School: Beijing Union Medical College
Course: Immunology
Keywords: umbilical cord mesenchymal stem cell tissue EGF bFGF
CLC: R329
Type: Master's thesis
Year: 2011
Downloads: 397
Quote: 0
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Abstract

Mesenchymal stem cells (MSCs) derived from mesoderm.They are multipotent somatic stem cells which can be induced into osteoplast, lipocyte as well as neuron. Human MSC show highly efficient homing to the dysfunctional tissue or organ. Meanwhile, MSC greatly reduce the clinical transplant rejection and improve the viability of clinical transplantation due to their low immunogenicity. Umbilical cord derived mesenchymal stem cells are more and more attractive as a possible alternative to bone marrow derived mesenchymal stem cells owing to better accessibility. Therefore, how to quick and effective isolate mesenchymal stem cells is becoming a more and more important issue.This reserch include two parts.First we demonstrate whether mesenchymal stem cells can be prcduced in large scale by explant culture.Then we observed the sucessful rate and growth condition isolated by tissue culture both in different time course and ater freezing.We cultured the tissue in D/F12 medium with 20%FBS.We observed the time and the yeild of primary cells.As results show that we get 4.03±0.97x104cells/cm in 17.33±0.84 days from all six samples within 24hours; 1.22±0.24x104cells/cm in 22.00±1.72 days during 24-48 time course; 1.01±0.22x104cells in 21.69±2.15 days during 48-72 time course.We also attained 2.08±0.30x104cells/cm in20.60±2.13 days after thawing by 3 weeks cyropreservation.We can conclude that the yield of primary cells decreased as the treatment time elongation.Second,we verified if EGF and bFGF could promote the attachment of primary cells.We alluded to the the growth condition of attached cells by EGF and bFGF,phenotype and cell cycle of cells after passaging.The result shows that the promoting time of cell attatching to medium are 51.42±4.38hours,38.37±2.96hours,32.25±1.85hours,33.61±1.56hours seperately.There is significant differences between different groups after 7 days culture.The phenotype of HLA-Ⅰ,CD105,CD73 are positive,HLA-Ⅱ,CD45,CD34 are negtive.All groups can be differentiate into osteoplast and adipocyte.Compared with control group,the S phase is higer. To sum up,we can obtained th MSCs by explant culture.There is relationship between yeild and treatment time.There is no big difference between cryopreservation group and non-cryopreservation group. EGF and bFGF play roles in promoting cells proliferation and attachment to medium.

Research on stem cells isolated and cultured human umbilical cord charging method

Abstract

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