Perfusion cell cultures generate higher viable cell densities (VCD) and volumetric productivity compared to fed-batch cultures. However, due to the limited availability of small-scale perfusion models, perfusion systems are seldom used to produce licensed biotherapeutics. This study evaluated two small-scale perfusion mimic protocols to bridge the research-to-production gap. Shake flasks and the ambr250 HT were used to compare centrifugation and in situ gravity settling protocols. The centrifugation protocol achieved a peak VCD >50 million cells/mL and is well-suited to media formulation and feeding strategy comparisons. The in situ gravity settling protocol achieved ∼20 million cells/mL and is well-suited to cell productivity and stability studies. The cell retention steps resulted in temporary DO and pH changes but did not affect the overall culture's health. Further, both protocols were able to recover from an imposed long-duration DO stress, although the centrifugation protocol cultures had higher cell specific oxygen consumption rates indicative of higher culture stress. Both protocols sustained the cultures for 39 days with stable cell specific productivities (∼27 pg/cell·day). Overall, this study demonstrated the feasibility of two economic small-scale perfusion mimic protocols in a standard small-scale bioreactor system, which could be translated to other multi-unit small-scale bioreactor systems. •Centrifugation perfusion mimic protocol enables media formulation comparisons.•Centrifugation perfusion mimic protocol assists feeding strategy comparisons.•In situ gravity perfusion mimic protocols evaluates cell productivity stability.•Cell specific oxygen uptake rates indicate increased stress for CHO cells.