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Long-term culture-initiating cells (LTC-IC) produced from CD34+ cord blood cells with limiting dilution method
OBJECTIVE: Even though much progress has been made in defining primitive hematologic cell phenotypes by using flow cytometry and clonogenic methods, the direct method for study of marrow repopulating cells still remains to be elusive. Long Term Culture-Initiating Cells (LTC-IC) are known as the most primitive human hematopoietic cells detectable by in vitro functional assays. METHODS: In this study, LTC-IC with limiting dilution assay was used to evaluate repopulating potential of cord blood stem cells. RESULTS: CD34 selections from cord blood were completed succesfully with magnetic beads (73,64%±9,12). The average incidence of week 5 LTC-IC was 1: 1966 CD34+ cells (range 1261–2906).CONCLUSION: We found that number of LTC-IC obtained from CD34+ cord blood cells were relatively low in numbers when compared to previously reported bone marrow CD34+ cells. This may be due to the lack of some transcription and growth factors along with some cytokines and chemokines released by accessory cells which are necessary for proliferation of cord blood progenitor/stem cells and it presents an area of interest for further studies.

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Authors: Gülderen Yanıkkaya Demirel, Tülin Budak-Alpdoğan, Sema Aktaş, Mahmut Bayık

Keywords: Kök hücre, kordon kanı, LTC-IC, kısıtlı dilüsyon, CFU kültürü, akan hücre ölçer

  • Year: 2010
  • Number: 4
  • Referance No: 35002

  • Figure 1. A summary of the LTC-IC by LDA method. FC: Flow cytometry, LDA: Limiting dilution assay, CAFC: Cobblestone area forming cells
  • Figure 2. CAFC and CFUs obtained from CD34+ cord blood cells are shown. CAFC were measured at week 5 and then cells were transferred to MethoCult, a semicellulose commercial media used for CFU assays. CFUs were counted on day 12 (week 7). A. Cobblestone Area Forming Cells (CAFCs) after week 5. B. CFU-GEMM: Colony Forming Unit-GranulocyteErythroidMonocyteMacrophage. C. BFU-E: Burst Forming Unit – Erythroid. D. Giemsa-stained BFU-E. E. A HPP-Q (High Proliferative Potential-Quiscent) colony. F. A BFU-E and CFU-GM are seen in a nearby area of the petri dish in MethoCult media

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