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HyStem-C First-Time User Guide


Introduction

If this is your first time using hydrogels, performing a simple bench-top experiment to familiarize yourself with how the hydrogel works is recommended on a pilot study before larger number of animals.


Bench-Top Gelation Test

Based on your planned animal experiments, decide on the cell concentration, medium type, and medium dilution that will be used for your experiments. Perform a gelation test with HyStem-C hydrogel kit as described in "Guidelines for 3-D Cell Encapsulation in HyStem-C Hydrogels for Cell Delivery Applications” to determine the available working time for injections. These three variables are dictated as follows:

  • Cell concentration depends on the cell type and length of time desired for tumor growth. (In general, tumor cells injected in HyStem-C produce tumors more rapidly and of more reproducible size than cells injected in medium alone.)
  • Medium dilution is based on required gelation time of HyStem-C and the amount of nutrients needed to keep cells alive prior to injection.

Pilot Animal Study

It is strongly recommended that researchers using HyStem-C conduct a pilot study using a small number of nude mice to determine the optimal experimental conditions for a given cell line. The following variables should be considered in the pilot study:

  • Cell type: HyStem-C has been successfully used for injection and growth of primary cells and thirty or more cell lines, including most common cancer lines.
  • Cell density: You may need to empirically determine the optimal density for a given cell line and injection location.
  • Injection volume: You may need to empirically determine the optimal volume for a given cell line and injection location.
  • HyStem-C dilution with medium: The dilution will affect the rate of gelation and may also affect the rate of tumor growth. See “Guidelines for 3-D cell encapsulation in HyStem-C hydrogels for xenograft applications” for recommendations on how to set up a pilot experiment. We also recommend performing an in vitro study using the same conditions as anticipated for the in vivo study to control for cell viability.