Method of profiling a cell population
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Abstract
The present invention relates to a method of profiling a cell population comprising a step of detecting the presence or absence of at least two biological markers in said cell population, wherein at least one of said markers is a cell surface marker, which is a sialylated N-glycan marker with structure NeuNAcα3Gal, and at least one of said markers is a mRNA marker related to glycoproteins and/or glycosynthase proteins. The invention also relates to method for purification of cord blood cell population and to a complete cell population from cord blood purified by said method.
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Citations
113 Claims
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1-72. -72. (canceled)
- 73. A method for purification of a cell population or a cord blood cell population from a sample containing multiple cell populations, comprising the following subsequent steps of contacting said sample with a specific binding reagent, removing material which is not bound to said specific binding reagent, contacting said sample with an additional amount of the specific binding reagent and removing material which is not bound to said specific binding reagent.
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85. A complete reproducible cell population purified by a binding reagent from a sample comprising multiple cell populations, wherein sample cell population comprises substantial amount of cells binding weakly to the binding reagent, the amount of weakly binding cells being between 0.1-5% or 5-50% of the cell population.
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86. A complete cell population wherein said cell population is from cord blood selected with regard to a single cell surface marker, said cell population being characterized by
a. purity of at least 90%; -
b. the presence of at least 50% of available cell surface marker containing cells of cord blood; and c. the presence of essentially natural ratio of cells expressing low and high levels of the surface marker. - View Dependent Claims (87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 110)
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- 97. A method of profiling a cell population of cord blood, preferably CD133-type homogenous cell population, comprising a step of detecting the presence or absence of at least two biological markers in said cell population, wherein at least one of said markers is a cell surface marker, and at least one of said markers is a mRNA marker, with the provision that said cell surface marker is not CD133 or CD34.
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111. A method for selecting optimal biomarkers for a stem cell population, preferably for a complete cord blood derived CD133 or CD34 cell population, the method comprising the steps of:
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a. producing a pure stem cell population; b. selecting a marker from markers listed in Tables 1-27 for testing the cell population; c. producing specific binding molecule recognizing the marker; d. testing the binding of the binding molecule to the cell population or testing the manipulation of the cell population by the binding molecule; e. optionally repeating the test with multiple cell populations from different individuals and with a control cell population(s); and f. selecting the optimal binding protein based on the binding activity and/or specificity and/or reproducibility of the protein.
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112. Method for combined mRNA expression analysis and glycome analysis, the method comprising the steps of:
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a. producing a pure stem cell population; b. screening mRNA expression levels of the cell population and screening of N-glycans of the cell population by carbohydrate recognizing screening methods; c. correlating the glycan expression with mRNA expression; and d. selecting the structure correlated in the previous step as a marker structure, wherein the cell population is preferably a complete cord blood derived CD133 or CD34 cell population.
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113. Method of assigning glycan structures with specific cell surface marker proteins, the method comprising the steps of:
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a. producing a pure stem cell population; b. isolating glycoprotein(s) from the cell; c. analyzing glycan structures of the isolated protein and/or the identity of the protein; wherein the cell population is preferably a complete cord blood derived CD133 or CD34 cell population.
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Specification