Embryonic stem cells defective in the pig-a gene show hematopoietic differentiation in vitro

V. Rosti, G. Tmmml, I. Uwatm, M. Bessler

Research output: Contribution to journalArticle

Abstract

Somatic mutations in Uie X-linked PIG-A gene m a hematopoiettc stem eel! are responsible fui the decreased expression of glycosylphospbaodyl inositol (GPI)-Iinked surface molecules on blood cells of patients with paroxysmal nocturnal hemoglobinuria (PNH). In order to study various features of blood cells devoid of GPl-linked proteins, we have induced erythroid and myetoid difterentiation inemoryoid bodies (EB) derived from wild type mouse embryonic stem (ES) cells and in EB derived from ES-cells in which an inactivating mutation m the pi$-a gene had been introduced by homologous recombination {pig-a null ES-cells). ES-cells were (a) plated directly in 0.9 mcthylceUulose supplemented with 15% Foetal bovine serum (FBS) and 1 U/ml erythropoietin or (b) first grown for 24 hours m liquid culture (with FBS bul without erythropoietin), whereby smalt cell aggregates were allowed to form, and then plated as above. Erythroid and/or myelotd differentiation was evaluated by light microscopy. The identity of cells was further confirmed by microscopic examination of cytospin preparations obtained from indivkfria) embryoJd bodiesofEB pctju of EB poMju ofEB pqgj Jiretily 247 S.I 16? 5 (>' 6 61 0 i . (1%| . |Q%) .the iU-41 After liquid 526 I4 115 22 156 73 I) cullute.Our data (summarized in the above table) show that (i) pig-a null ES-cells do form EB m vu m and (although in three separate experiments we observed some variability) these do differentiate into erythroid and myeloid cells; (ii) pig-a null ES-cd Is form less EB than wild type ES-cells, (iii) 24 hours liquid culture favours subsequent honatopoictic differentiation in EB derived from pi$-a null ES-cd Is. ht vitro dffercntution of ES-cd Is carrying a mutation of the pif-a gene will be useful to further investigate normal and PNH hematopoiesis and their response 10 environmental factors.

Original languageEnglish
Pages (from-to)1028
Number of pages1
JournalExperimental Hematology
Volume24
Issue number9
Publication statusPublished - 1996

ASJC Scopus subject areas

  • Cancer Research
  • Cell Biology
  • Genetics
  • Hematology
  • Oncology
  • Transplantation

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