In 1912, Bass and Johns described successful culture of
Plasmodium asexually in vitro [
1]; in 1976, Trager and Jensen achieved continuous in vitro cultivation of
Plasmodium falciparum [
2,
3], providing for the first time stable access to large quantities of stage specific malaria parasites [
2], including sexual stages, the so-called gametocytes [
4]. In search of an isolate for stable continuous in vitro culture and transmission experiments, the Nijmegen falciparum strain 54 (NF54) was selected. This was a case of airport malaria in a patient residing near Schiphol international airport in the Netherlands in 1979 [
5]. This isolate, thought to have originated from West-Africa [
6], and its clone 3D7 [
7] are arguably the most widely used laboratory strains of
P. falciparum and have become a standard resource for malaria research. They have formed the basis of whole sporozoite vaccination approaches [
8], and the basis of many functional assays for evaluation of malaria interventions, such as the standard membrane feeding assay (SMFA) [
9]. Given the importance of genetic variation for vaccine and drug efficacy, reliance on a single parasite isolate fails to give a comprehensive insight into intervention potency in natural infections [
10]. Similarly, relevant inter-strain variation in parasite growth rates [
11], gametocyte production [
12] and sporozoite invasion capacity [
13,
14] warrant further examination. The current portfolio of laboratory isolates for studies on sexual and sporogonic stages is very limited. During continuous culture, isolates can acquire spontaneous mutations and lose their ability to sexually differentiate [
15], hampering utility of many
Plasmodium isolates for malaria research. Expanding the portfolio to include a diversity of isolates would be beneficial. This study describes selection, clonal adaptation, and evaluation of
P. falciparum isolates for in vitro culture and sporogonic stage development.