Laboratory Adventures- Making Cell Culture Media

This week, we have been focusing on growing cells with the ST2 cell lineage. This cell lineage is one of the skeletal lineages, which means that these particular cells will eventually develop into a skeletal cell. What we want to do is get these cells to multiply, without having them differentiate into the specialized cells yet. Why? Because we want to have enough stem cells that can become the specific cells we want. Think of it like planting flowers. If you want to have a specific number of flowers in your garden, say 15, you need 15 seeds that will grow into those flowers. The same concept applies here with the skeletal stem cells: we need the right amount of stem cells that will become the right amount of bone.

So my next short story entails the cell-culture preparation of the specific cell culture media needed for the ST2 cells type to multiply:

Culture Dilemma

“Oh, come on, Mabel!” Walter wailed. “Please eat your electroplants! They are essential for keeping your organs magnetized!”

A white cow with magnetic silver splotches gave a loud bellow. She swiveled her head from the batch of sizzling, sparking white flowers at her left. They were eight feet tall, sprouting from a patch of orange soil encapsulated by a two-foot plastic fence.

“Ya’ll think I wanna be digestin’ that electrifying garbage?” The cow spoke through a series of deep moos and grunts (Walter was able to translate her guttural sounds into sentences). “Ya’ll don’t appreciate mah milk, so why should I go sufferin’ through the consumption of those junky products ya’ll call flowers?”

Mabel was a genopet species of a cow, known as a Magnecow. Such cows had magnetic organs. The milk they produced was known as “Magnemilk,” a special type of milk that contained bone marrow stromal cells—or BMSCs—which were stem cells that became bone. If one consumed the milk, one would consume the stem cells needed for proper bone growth, which meant rapid bone-healing or bone-growth once the stem cells in the milk were consumed. Such a quality was what made the Magnecows special: in the milk of many other genopet species of cows, the stem cells are missing because they mix with all the other ingredients in the milk, and they become dead and ineffective in the body. But since the Magnecows have magnetic organs, all the unwanted ingredients stick to the organs and the stem cells are still alive by the time Magnecow farmers like Walter obtain their milk.

Unfortunately, however, Walter’s 10-year-old son, Tommy, had refused to drink a single cup of her milk whenever his mother served it to him. He had been dumping it down the sink behind her back. To make matters worse, the other children of the Magnecow Clan had also been throwing away the Magnemilk cartons whenever they were served it in school.

Walter shook his head. “Kids will be kids. You can’t refuse to produce Magnemilk just because the children aren’t appreciating it. There are many other humans in this clan who love your milk! Look at all the muscular farmers! They depend on your Magnemilk!”

He put on his plastic gloves and grabbed the sparky, sizzling flowers at her left and shoved the flowers toward her jaws. The sparks surrounding the flower petals flickered, zapping the cow’s face. Mabel let out a loud bellow and whirled around, kicking her back leg toward Walter’s ribcage. Walter let out a quick grunt before everything went black.

“Walter, darlin’!” Ava found her husband lying on a white bed in a hospital, right next to their son, Tommy. She rushed up to his side and grabbed her cheeks with her hands. “Ain’t this a let-down. Can’t believe Mabel would do this to ya!”

He wrinkled his nose. “Hmph. Here I am with a broken ribcage. Now I need that genopet’s Magnemilk more than ever.”

Ava looked around the hospital, frowning. Everywhere she looked, there were doctors pushing stretchers in every direction, and Magnecows passing by with the doctors by their sides. “It’s pretty darn crowded here.”

“Well, our clan raises the cows that heal broken bones!” Snapped Walter. “Now that our Magnecows won’t eat any electroplants, their organs aren’t magnetized anymore, so they can’t produce any Magnemilk. Their milk will be just as lame and unhandy as any regular cow milk!”

Tommy groaned in the bed next to him. “Ohhhh, come on! Can’t you just make Magnemilk yourselves, without having to obtain it directly from Magnecows?”

Ava crossed her arms. Her eyes shifted from her son to her husband. Then she looked around at all the other patients in the hospital. Each patient either had deformed limbs or bones jutting out of place. She shuddered. She wasn’t a doctor in this hospital, but she was a postdoctoral researcher here.

“I’m gonna have a talk with my colleagues in my lab,” she spoke with a dry tone to her voice. “We’re gonna make sure we do just that,” she nodded to Tommy. “We’ll find a way to make Magnemilk without having to obtain it from Magnecows.”

Apparently, there was already a lab meeting to discuss a similar topic. Ava found herself in a room full of other researchers, each of them with silver hair just like her. They were all humans of the Magnecowish race, each of them responsible for raising Magnecows. Each was accompanied by their own Magnecows, just as Ava herself was accompanied by a cranky Mabel to her right.

However, there were other humans in the room as well. To their left was a group of humans with red hair, each accompanied by a cow with orange fur and flaming-red horns. They were the Flamecow Clan, and their cows were raised to provide hot milk tea that could cure cancer by burning off all cancerous cells. To their right was a group of humans with green hair, each accompanied by green-pelted goats with tan underbellies. They were the Goaticillin Clan, and their goats provided milk that was a mixture of penicillin and streptomycin, and their milk could cure pneumonia and a list of other respiratory diseases.

Mabel was shaking her head as she watched all the humans conversing amongst one another, shaking hands and exchanging small talk. Meanwhile, the three genopet species were all staying on their side of the room, snorting and glaring at one other.

“This is absurd,” mooed Mabel. “Can’t believe ya’ll are doin’ this.”

“Hey,” Ava shrugged. “You Magnecows didn’t want to eat any more electroplants, so we can’t obtain the milk to grow bones. Magnemilk was our only source of bone stem cells, but since you cows ain’t producing the milk anymore, we have to find another way to obtain enough bone stem cells that can turn into enough bone for all our injured patients.”

“Do you have to involve other clans in your pursuit?!” Mabel stomped her hoof. “You know we genopets abhor the scent of genopet species that are not our own!”

Ava sighed. “Hun, until we do any further research, this is the only way. By mixing Goaticillin milk, Flamecow milk, and regular, stem cell-derived Magnecow milk, we can create a cell culture media that will allow bone stem cells to grow and multiply. This is so that we have enough bone stem cells that can turn into bone once they are placed onto the right material.”

She smiled as a green-haired lady came up to her and handed her a bottle of dilute, tan liquid. Goaticillin milk, labeled as “Goaticillin Penstrep.” “Thank you,” she said.

The green-haired lady just wrinkled her nose and walked away.

An orange-haired man came up to her and tossed a bottle of thicker orange liquid. Flamecow milk, labeled as “Flamecow MEM-alpha.”

“Whoa!” Ava caught the bottle, though it almost slipped out of her hands. “Sheesh, careful there!”

Mabel snorted at the man. “Pah! No one seems to be happy to work with ya’ll.”

“Well, we paid ‘em a ton of money to obtain this stuff,” she nodded at both bottles of milk in her hands. “This better work.”

She let Mabel into her lab room. She milked Mabel into a small bottle, labeled “Magnecow FBS.” It was short for “Magnecow Fetal-Bone-Serum.”

She poured each of the three milk bottles into a 50-mL beaker and then turned to three petri dishes. Each was plated with bone stem cells. She pipetted the solution from the beaker into three petri dishes. Then she placed them in the incubator.

She didn’t return until three days later. When she opened the incubator to check the cells under the microscope, she saw numerous clusters of tiny dots, crowded together.

She clapped her hands. “They’ve grown! The bone stem cells have grown!”

Mabel awoke from her stall in the back of the lab. “Ay…what’s all the yellin’?”

“Mabel, it’s a miracle! Using regular milk from you Magnecows and mixing it with Flamecows and Goaticillins made the right media to make these bone stem cells multiply. Just pouring it over three petri dishes has made enough bone stem cells to grow an entire femur for my son!”

Mabel chortled. “Heh. So now you’re just gonna use that milk-mixin’ method? Good luck with your future debt to the Flamecow and Goaticillin Clans.”

——–

Notice the title of this short story was not “Cultural” Dilemma, but “Culture” Dilemma. Culture referred to both the cell culture we were trying to prepare, as well as the different clan cultures that were needed to provide the resources for it!

But the main correlation to my experiment was the way the cell culture media had to be prepared. To prepare cell culture media for the ST2 cells, I had to mix MEM-alpha, which is base media, along with fetal bovine serum (FBS), and a solution of penicillin and streptomycin, which is known as “penstrep” for short. Mixing those three ingredients would create the complete cell culture media for the ST2 cell lines, so that these cells can continue to multiply until enough is needed for the next step.

Then later on in the week, we plated these ST2 cells onto the wooden PEEK material. Unfortunately, the cells didn’t stick as well as we wanted them to. So for this upcoming week, we will focus on obtaining different skeletal lineages. The plan is to isolate the cells from bone marrow, and then put the entire cell population into a flask. After about a day, there may be cells that stick to the flask. These are the cells that possibly are of skeletal lineage. We will take the cells that stuck to the flask and use flow cytometry to decipher the specific types of cells and see if we have obtained the cells we want. Stay tuned for my next short story, which will entail more details on how this experiment will go!

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One Response to Laboratory Adventures- Making Cell Culture Media

  1. Parker J says:

    I wasn’t sure if you could pull this off but you did. Good for you! Looking forward to more.

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