Applying Excel Spreadsheet for Guide, Process Control, Documentation, and Labeling Assistance for HPCA Cryopreservation

Background \u0026 Aim HPCA plasma depletion and cryopreservation is one of the most common processes in a cell processing facility. The process steps are:(1)initial sampling, evaluation of the in-coming WBC concentration, volume, CD34/kg, (2)centrifugation, (3)plasma depletion, (4)aliquot the plasma depleted product into cryo freezing bags, (5)adding cryoprotectant(DMSO based),(6)post-process sampling, (7)controlled rate freezing. There are several critical values during the process:(a) the plasma depletion rate is the higher the better, to reduce the use of DMSO, while(b) keeping the WBC recovery rate as high as possible (minimum 75%, usually \u003e90% in our facility), and (c)the final product WBC concentration not too high (should not exceed 560e6/ml in our facility); also considering (d) the filling volume of the cryo freezing bags per manufacturer\u0027s specification, which is 30-70mL/bag in our facility; (e)CD34/kg per bag requirement per \"Processing Order\", which is 2e6 in our facility. The process should be performed as quickly as possible to maximize cell viability. Since there are so many numbers to consider during the process, manual calculation could be slow and/or inaccurate, especially under time pressure. Therefore, our facility developed and validated an Excel spreadsheet with multiple automatic calculation functions to guide the critical steps of cryopreservation. also as a documentation and a labeling tool. Methods, Results \u0026 Conclusion An excel spreadsheet with \"Initial, In-Process, Aliquot Suggestion, Actual, Final Product\" sections, is developed, encompassing the above required data and limiting values. It used many \"IF\" functions in formula, conditional formatting, data validation (the entry cells will either \"warm\" or \"stop\" if non-logical data entered; certain cells will flag a red X if undesirable data achieved). It can suggest the operator to deplete how much plasma, and to suggest how many cryo bags to use. The operator can make decision and enter in the \"actual\" cells, the spreadsheet will calculate the actual volumes in each bag accordingly. An in-process control point where the TNC recovery rate is estimated, after the plasma depletion. The spreadsheet is trialed, modified, finally validated per our Worksheet Validation SOP. The spreadsheet is password protected. See below image of the spreadsheet and formula provided for downtime purpose. The spreadsheet has facilitated the quality, speed, and safety of the cryo process, also assisting the documentation and labeling process.