Reduced growth of normal myeloid progenitors on MDS stroma is unrelated to confluence

Tennant, G. B., Truran, Louise N. and Burnett, Alan Kenneth 1996. Reduced growth of normal myeloid progenitors on MDS stroma is unrelated to confluence. British Journal of Haematology 93 (S2) , p. 66. 10.1111/j.1365-2141.1996.tb08988.x

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Abstract

Ineffective haemopoiesis in patients with myelodysplasia (MDS) may be a consequence of defective marrow stromal cells. We have measured the growth of normal haemopoietic progenitors from cord blood mononuclear cells (MNC) in secondary long-term cultures on irradiated stromal layers grown from the marrow of normal subjects and MDS patients. Stromal layers were established in 25 cm2 flasks by inoculating 20 x 1 O6 bone marrow MNC in 10 ml of modified McCoy's 5A medium (glucose reduced to 400 mg/l and supplemented with 450 mg galactose/l) buffered with 40 mM HEPES.The proportion of the flask surface covered by stromal cells was estimated after 5 weeks primary growth (confluence %) and layers irradiated (10 Gy). Secondary cultures were established by re-inoculating with 5 x lo6 MNC from cord blood. Confluence ranged from 60-100% in normal cultures (n = 25; median = 95%) and 40-100% in MDS cultures (n = 26; median = 90%; ns). After 5 weeks secondary culture the CFU-GM from cord blood had a median value of 58.0/culture when grown on normal stroma (n = 20) and 4.3 on MDS stroma (n = 19; p = 0.0002). Median erythroid colonies numbered 15/culture with normal stroma (n = 13) and 0 with MDS stroma (n = 19; p = 0.004). In cultures with 90% confluence, or more, the median CFU-GM were 98.9hormal culture (n = 13) and 6.1IMDS culture (n = 12; p = 0.002); and the median period of CFU-GM detectability was 10 weeks on normal stroma (n = 9) and 5 weeks on MDS stroma (n = 13; p = 0.0005). There was little evidence of correlation between confluence and secondary CFU-GM numbers in weeks 4 to 7; but the period of CFU-GM detectability was correlated to confluence on normal stroma (n = 16; r, = 0.6463; p = 0.006) but not on MDS stroma (n = 20; r, = 0.2964; ns). Mean culture pH values were identical at week 5 (normal = 7.13, n = 23; MDS = 7.13, n = 19; ns) suggesting there was no difference in the underlying metabolic activity. We conclude that stromal layers from MDS marrow are defective in their ability to support normal haemopoiesis; and the variation in this defectiveness is greater than the measured variation in confluence and is independent of it.

Item Type:	Article
Date Type:	Publication
Status:	Published
Schools:	Medicine
Subjects:	R Medicine > R Medicine (General)
Additional Information:	Abstracts of papers presented at the Second Meeting of the European Haematology Association, Paris, France, 29 May-1 June 1996. Supplement 2, Article 260.
Publisher:	Wiley-Blackwell
ISSN:	0007-1048
Last Modified:	18 Sep 2017 09:49
URI:	https://orca.cardiff.ac.uk/id/eprint/59199

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