Chlorella is one of the common microalgae found in a wide range of habitats, including Antarctica. Chlorella UMACC 234 is an interesting isolate in the collection of Antarctic microalgae in the University of Malaya algae culture collection (UMACC) as it grows well at temperatures much higher than the ambience. The alga was isolated from snow samples collected from Casey, Antarctica. This study investigates the influence of nitrogen source on the growth, biochemical composition and fatty acid profile of Chlorella UMACC 234. The cultures were grown in Bold’s Basal Medium with 3.0 mM NaNO3, NH4Cl or urea. The cultures grown on NaNO3 attained the highest specific growth rate (μ = 0.43 day–1) while the specific growth rates of those grown on NH4Cl and urea were not significantly different (p > 0.05). The urea-grown cells produced the highest amounts of lipids (25.7% dry weight) and proteins (52.5% dry weight) compared to those grown on other nitrogen sources. The cell numbers attained by the cultures grown at NaNO3 levels between 0.3 and 3.0 mM were similar but decreased markedly at 9.0 mM NaNO3. The fatty acids of Chlorella UMACC 234 were dominated by saturated fatty acids, especially 16:0 and 18:0. The percentage of polyunsaturated fatty acids was very low, especially in cells grown on urea (0.9% total fatty acids). Characterisation of the growth and biochemical composition of this Antarctic Chlorella is important to our studies on the relationship of Chorella isolates from tropical, temperate and polar regions, especially in terms of phylogeny and stress adaptation.