Effects of FPIL6/IL2 on the differentiation and proliferation of lymphoid in the ML-IC assay.
- Author:
Er-jin FAN
1
;
Jun-min SONG
;
Shi-rong XU
;
Chun-hua ZHAO
Author Information
- Publication Type:Journal Article
- MeSH: Animals; Biological Assay; Cell Culture Techniques; methods; Cell Differentiation; Cell Division; Cells, Cultured; Coculture Techniques; methods; Hematopoietic Stem Cells; cytology; Humans; Interleukin-2; biosynthesis; genetics; pharmacology; Interleukin-6; biosynthesis; genetics; pharmacology; Killer Cells, Natural; cytology; metabolism; Mice; Recombinant Fusion Proteins; pharmacology; Stromal Cells; cytology
- From: Acta Academiae Medicinae Sinicae 2002;24(1):30-35
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo develop an in vitro assay that allows the culture and identification of a single human bone marrow progenitor closely related to hematopoietic stem cell, which is more primitive than LTC-IC, and to find an efficient culture conditions for NK-IC expansion.
METHODSFusion protein IL6/IL2 was reconstructed and expressed in E. coli DH5 alpha. ML-IC was determined by watching if the single cell can give rise to secondary progenitors with both LTC-IC and NK-IC characteristics. LTC-IC frequency was determined by the CFC clonogenic methylcellulose assay. NK-IC frequency was determined by phenotyping CD56 positive NK cells. The effect of FPIL6/IL2 on the expansion of NK-IC was examined by comparing the colony number of NK cells before and after the culture.
RESULTSAfter the initial 4-week expansion culture, we showed that (25.75 +/- 5.68)% of freshly sorted Lin-/34+/DRdim cells were able to generate functional NK-IC in one or more secondary FPIL6/IL2 cultures, whereas (6.81 +/- 1.97)% in the control. A total of 102 NK-IC cells were present when were cultured for 6-7 weeks in FPIL6/IL2 expansion medium, which was much higher than the 33 NK-IC cells in the control.
CONCLUSIONML-IC assay will prove useful to assess a very primitive hematopoietic cell with multilineage generative capacity. FPIL6/IL2 is capable of initiating and promoting NK-IC expansion greatly in ex vivo cultures in terms of net-conservation and net proliferation.
